Clostridioides difficile infection (CDI) may recur in approximately 10–30% of patients, and the risk of recurrence increases with each successive recurrence, reaching up to 65%. C. difficile can form biofilm with approximately 20% of the bacterial genome expressed differently between biofilm and planktonic cells. Biofilm plays several roles that may favor recurrence; for example, it may act as a reservoir of spores, protect the vegetative cells from the activity of antibiotics, and favor the formation of persistent cells. Moreover, the expression of several virulence genes, including TcdA and TcdB toxins, has been associated with recurrence. Several systems and structures associated with adhesion and biofilm formation have been studied in C. difficile, including cell-wall proteins, quorum sensing (including LuxS and Agr), Cyclic di-GMP, type IV pili, and flagella. Most antibiotics recommended for the treatment of CDI do not have activity on spores and do not eliminate biofilm. Therapeutic failure in R-CDI has been associated with the inadequate concentration of drugs in the intestinal tract and the antibiotic resistance of a biofilm. This makes it challenging to eradicate C. difficile in the intestine, complicating antibacterial therapies and allowing non-eliminated spores to remain in the biofilm, increasing the risk of recurrence. In this review, we examine the role of biofilm on recurrence and the challenges of treating CDI when the bacteria form a biofilm.

To determine genomic characteristics and molecular epidemiology of carbapenem non-susceptible Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, and Pseudomonas aeruginosa from medical centres of Mexico using whole genome sequencing data analysed with the EPISEQ^Ⓡ CS application and other bioinformatic platforms. Clinical isolates collected from 28 centres in Mexico included carbapenem-non-susceptible K. pneumoniae (n = 22), E. coli (n = 24), A. baumannii (n = 16), and P. aeruginosa (n = 13). Isolates were subjected to whole genome sequencing using the Illumina (MiSeq) platform. FASTQ files were uploaded to the EPISEQ^Ⓡ CS application for analysis. Additionally, the tools Kleborate v2.0.4 and Pathogenwatch were used as comparators for Klebsiella genomes, and the bacterial whole genome sequence typing database was used for E. coli and A. baumannii. For K. pneumoniae, both bioinformatic approaches detected multiple genes encoding aminoglycoside, quinolone, and phenicol resistance, and the presence of bla_NDM-1 explained carbapenem non-susceptibility in 18 strains and bla_KPC-3 in four strains. Regarding E. coli, both EPISEQ^Ⓡ CS and bacterial whole genome sequence typing database analyses detected multiple virulence and resistance genes: 20 of 24 (83.3%) strains carried bla_NDM, 3 of 24 (12.4%) carried bla_OXA-232, and 1 carried bla_OXA-181. Genes that confer resistance to aminoglycosides, tetracyclines, sulfonamides, phenicols, trimethoprim, and macrolides were also detected by both platforms. Regarding A. baumannii, the most frequent carbapenemase-encoding gene detected by both platforms was bla_OXA-72, followed by bla_OXA-66. Both approaches detected similar genes for aminoglycosides, carbapenems, tetracyclines, phenicols, and sulfonamides. Regarding P. aeruginosa, bla_VIM, bla_IMP, and bla_GES were the more frequently detected. Multiple virulence genes were detected in all strains. Compared to the other available platforms, EPISEQ^Ⓡ CS enabled a comprehensive resistance and virulence analysis, providing a reliable method for bacterial strain typing and characterization of the virulome and resistome.

This review explores the role of Pseudomonas aeruginosa biofilms in cystic fibrosis (CF) pathogenesis. Biofilms, the main bacterial lifestyle in CF lungs, are key in therapy failure, immune evasion, and chronic infection persistence. This review examines biofilm structure, emphasizing extracellular polymeric substances (Psl, Pel, alginate, eDNA) and their roles in structural stability, resistance to antibiotics, and immune modulation. Regulatory mechanisms, including c‐di‐GMP signaling and quorum‐sensing systems, are detailed as key drivers of biofilm formation and maintenance. The review also highlights polymicrobial interactions, particularly with Staphylococcus aureus , Candida spp., and Aspergillus spp., and commensal bacteria, illustrating how interaction dynamics shape microbial behavior, virulence, and treatment outcomes. Methods for studying biofilms in CF‐like conditions, such as advanced in vitro models and transcriptomic analyses, are outlined for their relevance in replicating the complex lung environment. Emerging antibiofilm strategies, including matrix‐disrupting enzymes, quorum‐sensing inhibitors, bacteriophage therapies, and nanomedicine, are discussed as promising tools to combat biofilm resilience. The review underscores the need for innovative therapeutic approaches and a deeper understanding of microbial and host interactions to improve clinical outcomes in CF patients.