Lidia Piechowicz

Background/Objectives: Shiga toxin-producing Escherichia coli (STEC) are important zoonotic pathogens associated with gastrointestinal infections and hemolytic–uremic syndrome (HUS). Increasing serotype diversity, emergence of hybrid strains, and growing antimicrobial resistance complicate surveillance and treatment. This review summarizes recent data on STEC isolated from major animal reservoirs between 2020 and 2026. Methods: We comparatively analyzed peer-reviewed, open-access studies investigating STEC from cattle, sheep, goats, poultry, wildlife, and related animal products. We extracted and evaluated data on serotypes, virulence genes, hybrid pathotypes, and antimicrobial resistance determinants. Results: High serotype diversity was observed across all reservoirs, with non-O157 STEC frequently predominating over O157:H7. Ruminant isolates commonly carried stx2 and additional virulence determinants associated with severe human disease, confirming cattle and small ruminants as major reservoirs of highly pathogenic STEC. Poultry-associated isolates showed greater variability in virulence profiles but frequently exhibited high rates of multidrug resistance. Hybrid strains combining virulence traits characteristic of multiple E. coli pathotypes were increasingly reported. Livestock-associated STEC commonly carried resistance determinants against β-lactams, tetracyclines, sulfonamides, quinolones, and polymyxins, whereas wildlife isolates generally showed lower rates of resistance. Conclusions: Animal-associated STEC populations demonstrate increasing genomic diversity, pathogenic potential, and antimicrobial resistance. The growing prevalence of non-O157 and hybrid strains highlights the limitations of traditional serotype-focused surveillance and emphasizes the need for integrated One Health monitoring strategies.

Background/Objectives: Skin and soft tissue infections (SSTIs) represent a significant clinical challenge, largely due to the high prevalence of antibiotic-resistant Staphylococcus aureus, particularly methicillin-resistant S. aureus (MRSA). Treatment is further complicated by biofilm formation, which reduces antibiotic efficacy. The limitations of conventional therapies highlight the need for alternative approaches. Phage therapy has emerged as a promising biological strategy; however, its effectiveness may be constrained by factors such as phage instability and biofilm regrowth. This study aimed to enhance phage-based treatment by combining a newly isolated phage, vB_Sau-E, with lactoferrin (Lf), a multifunctional protein of the innate immune system. Methods: Phage vB_Sau-E was characterized in terms of its infection dynamics and lytic activity. Biocompatibility was further examined using human skin cell lines. The potential effect of Lf was assessed by evaluating its impact on phage infectivity and stability under a range of environmental conditions and by checkerboard assay. Results: Phage vB_Sau-E belongs to the Silviavirus genus in the Herelleviridae family. It was shown to infect 12 out of 22 tested clinical MRSA isolates, with 10 strains identified as good hosts. The phage has a ~30 min life cycle, and ~50 progeny virions are released after bacterial cell lysis. We have also observed that Lf increased plating efficiency and enhanced phage stability at a pH of 5.5 and at −20° C. It also proved to have an additive antibacterial effect, though this was observed to be strain-dependent. Conclusions: Lactoferrin functions as a stabilizing adjuvant for phage vB_Sau-E. Its additive effect supports the development of more effective, biofilm-targeting therapies for staphylococcal SSTIs.

Bacterial resistance to antibiotics is one of the leading factors encouraging the development of new therapeutic strategies. The increased resistance to antibiotics can be attributed to several factors, such as early and unnecessary administration, incorrect dosing, or incomplete antibiotic treatment. One of the diseases that calls for improved understanding of this problem is meningitis, which - if ineffectively treated - may result in severe neurological complications and death. This study provides an overview of the current antibiotic strategies for bacterial meningitis along with the therapeutic challenges associated with standard treatment options. In addition, it also presents the current progress in bacteriophage research, highlighting both their potential to replace some common antibiotic therapies in the treatment of meningitis and the significant lack of clinical studies regarding most of them. The research on phage therapy targeting meningitis-associated pathogens is limited, and where it exists, it is predominantly focused on mouse models. There, its efficiency seems mostly promising. Nevertheless, comprehensive clinical trials are needed to properly determine the efficacy and safety of phage therapy in humans before it becomes a significant alternative to antibiotics.

Staphylococcus aureus nasal carriage is a potential source of secondary infections in COVID-19 patients, yet it remains unclear whether SARS-CoV-2 infection favors colonization by more virulent or resistant strains. We analyzed 31 nasal S. aureus isolates from hospitalized COVID-19 patients to assess antimicrobial resistance, virulence gene content, and genetic diversity. Only two isolates (6.4%) were methicillin resistant, and most strains showed limited resistance beyond the MLSB phenotype. Adhesin genes were highly prevalent, whereas toxin genes were detected in only 16.1% of isolates. Spa typing revealed high genetic diversity with no dominant clone. Overall, S. aureus isolates from COVID-19 patients did not differ substantially from previously described carriage strains, suggesting no selective enrichment of highly virulent or resistant lineages during SARS-CoV-2 infection.

Staphylococcus aureus is a major cause of hospital-acquired pneumonia, with methicillin-resistant strains contributing significantly to prolonged illness and mortality. Methicillin-resistant strains can be responsible for up to 75% of infections in certain countries. Therefore, the problem is described as severe, and the search for alternative methods of treatment of such infections is currently one of the priorities in healthcare. Bacteriophages, although historically underutilized, are re-gaining interest for their potential in treating bacterial infections. However, they do have their limitations such as specific ranges of activity and resistance development. Combining phages with antimicrobial agents such as lactoferrin—a natural protein with antimicrobial and anti-biofilm properties—may improve treatment outcomes. In this study, we evaluated the efficacy of three Kayviruses paired with lactoferrin against MRSA in infected pulmonary epithelial cell cultures. The combination significantly reduced bacterial viability, protected human cells from cytotoxic effects of bacterial infection, and decreased inflammasome activation. These findings suggest that phage-lactoferrin combinations may offer a promising, safer alternative for managing MRSA-related pneumonia and reducing dependence on traditional antibiotics. Key points: • Phage lactoferrin mixture had no influence on A549 cells • Lactoferrin increased phage efficacy and reduced influence of bacteria on cells • Phage + Lf mixture limited inflammatory response similarly to phages and Lf alone

The impact of tetracycline on the soil and rhizosphere microbiome of lettuce was analyzed. Soil was collected from an agricultural field regularly fertilized with manure, and tetracycline was added at two concentrations (5 mg/kg and 25 mg/kg). In untreated soil, dominant bacteria included Proteobacteria (43.17%), Bacteroidota (17.91%), and Firmicutes (3.06%). Tetracycline addition caused significant shifts in the microbiome composition, notably increasing Actinobacteriota (22%) and favoring Mycobacterium tuberculosis (low concentration) and Mycobacterium holsaticum (high concentration). Proteobacteria decreased by 21%, possibly indicating antibiotic resistance development. An increase in Firmicutes, particularly Bacillales, suggested a selection for resistant strains. In the lettuce rhizosphere, tetracycline-induced changes were less pronounced than in soil. Proteobacteria remained dominant, but taxa like Burkholderiales and Chitinophagales increased in response to tetracycline. The rise in chitin-degrading bacteria might result from fungal overgrowth linked to the bacteriostatic effects of tetracycline. Pathogens such as M. tuberculosis, observed in the soil, were not detected in the lettuce rhizosphere.

Background/Objectives: The viral pandemic caused by the SARS-CoV-2 virus has affected millions of people. However, it was noticed that high mortality was often a result of bacterial co-infections. One of the main pathogens responsible for secondary infections in patients with viral respiratory tract infections, including COVID-19, is Staphylococcus aureus. In recent years, the number of infections caused by drug-resistant strains of S. aureus has been growing rapidly, often exceeding the number of infections caused by antibiotic-sensitive strains. In addition, biofilm-related infections are more difficult to treat due to the lower sensitivity of biofilm structure to antibiotics. Bacteriophages are seen as alternative treatment of bacterial infections. Therefore, in our work, we have analyzed the efficacy of three Kayviruses against S. aureus strains isolated from COVID-19 patients. Methods: We analyzed the ability of tested phages to remove S. aureus biofilm both from polystyrene plates as well as from the surface of pulmonary epithelial cells. Results: We have observed that tested Kayviruses had a broad host range. Furthermore, phages were able to effectively reduce biofilm biomass and number of viable cells in pure culture. During our research, none of the tested phages was shown to have a negative effect on cell viability and were able to inhibit the negative effect S. aureus had on cell condition. Conclusions: Our results show tested phages were effective in reducing the biofilm of S. aureus strains isolated from COVID-19 patients, had no adverse effect on lung epithelial cell viability. Therefore, it should be recognized that the properties of three studied Kayviruses give them an advantage in the selection of phages for treatment of staphylococcal infections.

Background/Objectives: According to the One Health concept, wild birds can be indicators of ecosystem pollution and disease incidence. Escherichia coli strains are widespread worldwide, but there are still few reports on the association of human infections with a potential reservoir of highly pathogenic human strains in wild birds. Fecal E. coli with uropathogenic potential (UPEC) can be transmitted between birds and humans and may be a risk factor for urinary tract infections (UTIs). Results: The results showed that above 50% of the isolates were grouped as highly pathogenic, according to Clermont phylogroup classification. Such strains were found to be stronger biofilm producers, with a higher adherence of monocytes than low pathogenic. However, the highest cytotoxicity was observed for strains described as aquatic environmental. Convergence of the results of the analysis of monocyte activation by E. coli strains and the ability to form biofilm by individual phylogroups of the strains tested was demonstrated. Genetic determinants of the uropathogenicity of E. coli (UPEC) correlate with the evidence of strain pathogenicity during monocyte activation in in vitro assays. Methods: In this study, we assessed the virulence potential of environmental strains isolated from wild waterfowl using genetic analysis (Clermont phylogroup classification) and phenotypic methods, including analysis of the human monocyte response to biofilm formation. The estimation of the ability to form biofilms was tested using crystal violet, and the pathogenic potential of strains by monocyte activation assay including changes in morphology, adhesion and cytotoxicity. Conclusions: In conclusion, the virulence of E. coli strains isolated from free-living birds is significant, and they can be considered environmental reservoirs of pathogenic strains. According to our observations, they can be responsible for the dissemination of uropathogenic strains among humans.

Vancomycin (Van) is a glycopeptide antibiotic commonly used as a last resort for treating life-threatening infections caused by multidrug-resistant bacterial strains, such as Staphylococcus aureus and Enterococcus spp. However, its effectiveness is currently limited due to the rapidly increasing number of drug-resistant clinical strains and its inherent cytotoxicity and poor penetration into cells and specific regions of the body, such as the brain. One of the most promising strategies to enhance its efficacy appears to be the covalent attachment of cell-penetrating peptides (CPPs) to the Van structure. In this study, a series of vancomycin conjugates with CPPs—such as TP10, Tat (47–57), PTD4, and Arg₉—were designed and synthesized. These conjugates were tested for antimicrobial activity against four reference strains (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa) and two clinical drug-resistant strains: methicillin-resistant S. aureus and vancomycin-resistant E. faecium. In addition, cytotoxicity tests (using a human fibroblast cell line) and blood–brain barrier (BBB) permeability tests (using a parallel artificial membrane permeability assay—PAMPA-BBB assay) were conducted for selected compounds. Our research demonstrated that conjugation of Van with CPPs, particularly with Tat (47–57), Arg₉, or TP10, significantly enhances its antimicrobial activity against Gram-positive bacteria such as S. aureus and Enterococcus spp., reduces its cytotoxicity, and improves its access to brain tissues. We conclude that these findings provide a strong foundation for the design of novel antimicrobial agents effective in treating infections caused by drug-resistant staphylococcal and enterococcal strains, while also being capable of crossing the BBB.