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Replacement of native joints aims to restore patients’ quality of life by relieving pain and improving joint function. While periprosthetic joint infection (PJI) affects a small percentage of patients, with an estimated incidence of 1–9% following primary total joint replacement, this postoperative complication necessitates a lengthy hospitalisation, extended antibiotic treatment and further surgery. It is highlighted that establishing the correct diagnosis of periprosthetic infections is critical in order for clinicians to avoid unnecessary treatments in patients with aseptic failure. Of note, the PJI diagnosis could not purely rely upon clinical manifestations given the fact that heterogeneity in host factors (e.g., age and comorbidities), variability in infection period, difference in anatomical location of the involved joint and discrepancies in pathogenicity/virulence of the causative organisms may confound the clinical picture. Furthermore, intra-operative contamination is considered to be the main culprit that can result in early or delayed infection, with the hematogenous spread being the most prevalent mode. To elaborate, early and hematogenous infections often start suddenly, whereas chronic late infections are induced by less virulent bacteria and tend to manifest in a more quiescent manner. Last but not least, viruses and fungal microorganisms exert a role in PJI pathogenesis.

The One Health approach emphasizes the importance of antimicrobial resistance (AMR) as a major concern both in public health and in food animal production systems. As a general classification, E. coli can be distinguished based on the ability to cause infection of the gastrointestinal system (IPEC) or outside of it (ExPEC). Among the different pathogens, E. coli are becoming of great importance, and it has been suggested that ExPEC may harbor resistance genes that may be transferred to pathogenic or opportunistic bacteria. ExPEC strains are versatile bacteria that can cause urinary tract, bloodstream, prostate, and other infections at non-intestinal sites. In this context of rapidly increasing multidrug-resistance worldwide and a diminishingly effective antimicrobial arsenal to tackle resistant strains. ExPEC infections are now a serious public health threat worldwide. However, the clinical and economic impact of these infections and their optimal management are challenging, and consequently, there is an increasing awareness of the importance of ExPECs amongst healthcare professionals and the general public alike. This review aims to describe pathotype characteristics of ExPEC to increase our knowledge of these bacteria and, consequently, to increase our chances to control them and reduce the risk for AMR, following a One Health approach.

Dairy herds around the world are undergoing several changes. Herd sizes are increasing, as are both milk yield and quality. The implementation of new technologies in various domains of dairy production is leading to an increase in the quantity of data available. This, in turn, creates a need to extract useful information from these data to improve production efficiency. This paper presents the findings of a preliminary study that utilizes a machine learning (ML) approach to assess the accuracy of somatic cell count (SCC) and neutrophils + lymphocytes count/mL (PLCC) in identifying cows at risk of developing intramammary infection (IMI) due to major pathogens. These pathogens (MajPs) include S. aureus, S. agalactiae, S. uberis, and S. dysgalactiae. This study identified these pathogens either by real-time PCR (qPCR) methods or by conventional bacteriology, following the cows’ calving process. This study encompassed a total of 424 cows and 1696 quarter milk samples. A comparison of the two methods revealed significant disparities in the prevalence of MajPs, with the qPCR method demonstrating a higher prevalence than conventional bacteriology. However, the prevalence of negative results was comparable, with both methods yielding approximately 71.0% and 72.1%, respectively. The comprehensive results of this study substantiated that all the cellular markers exhibited the most accurate when MajP IMI was diagnosed using quarter milk samples, but this result is mainly due to the very high specificity. The cellular markers exhibited nearly equivalent performance, irrespective of the ML algorithm employed. The findings indicate that approaches based on SCC or PLCC may be useful for identifying healthy cows or quarters. However, it is essential to confirm all “non-negative” results through subsequent analysis within 7–15 days to ensure accuracy. However, further studies are necessary to enhance diagnostic accuracy.

A customizable 3D-bioprinted core-in-shell platform was developed for time-dependent oral colon delivery of live microorganisms. The system conveyed as a model bacterial species within a monolithic core, which was surrounded by a swellable hydroxypropyl cellulose barrier, imparting a lag phase of programmable duration, and by an enteric outer layer, protecting the dosage form during unpredictable gastric residence. Pastes of different compositions were investigated to shape the core. Core and core-in-shell units were fabricated from digital models using a bioprinter equipped with a high-precision plunger dispenser and pressure-based thermoplastic printhead. The printed units were characterized in terms of mass, dimensions, mechanical properties and release performance using paracetamol as a reference tracer. Bacterial viability was evaluated during screening of the formulation components and after each processing step by manual counting of colony-forming units. A mannitol-based formulation was selected for fabrication of the core, offering a favorable balance of printability, physico-technological properties, release behavior and ability to preserve bacterial viability. Two-layer core-in-shell systems were manufactured via a dual-printing operating mode. The desired performance was attained, with no release under acidic conditions, a lag phase in pH 6.8 fluid and a subsequent release profile comparable with that generated by the core as such. Viability studies demonstrated that compounding, core printing, shell deposition and drying did not adversely affect survival. 3D bioprinting was proved to be a versatile technique for the manufacturing of oral colon delivery systems containing probiotics or live biotherapeutics.

Antimicrobials are common drugs used to treat and prevent infectious diseases in plants, animals, and humans. Since their discovery in the mid-20th century, their use has dramatically increased for the benefit of humanity, and also for animal care. However, antimicrobial resistance soon appeared, which, according to the WHO, will limit or impede their use at the horizon of 2050. Indeed, antimicrobial resistance (AMR), which is a natural phenomenon in bacteria increased dramatically over the last 3 decades mainly due to the overuse and misuse of antibiotics in humans, animals, and plants. Apart from affecting human health, drug-resistant diseases also adversely affect plant and animal health, reduce agricultural productivity, and threaten food security. AMR affects all countries, regardless of economic status, and imposes high costs on health systems and national economies. Therefore, antimicrobial resistance should be studied and analyzed under the One Health paradigm. In mind of the One Health paradigm, to reduce and overcome AMR, we must take at least 3 complementary and integrated actions: (i) monitoring the resistome; (ii) developing protective strategies against antibiotic resistance; (iii) taking curative actions by designing new and original treatments. Moreover, the three actions must be conducted simultaneously due to the continuous adaptation of bacteria.

Background/Objectives: Pyometra is a life-threatening uterine infection of intact bitches and queens. Despite growing reports of multidrug-resistant (MDR) Escherichia coli in canine reproductive and urinary infections, no whole-genome data were previously available for pyometra isolates from Italy. This study aimed to characterise, by whole-genome sequencing and comparative genomics, the population structure, resistome and virulome of E. coli causing pyometra in companion animals from northern Italy in the context of European datasets. Methods: Four E. coli isolates (two canine, two feline) from pyometra cases underwent nanopore long-read sequencing. Genomes were compared with Brazilian and Finnish pyometra isolates using core- and accessory-genome analyses, pan-genome partitioning, phylogeny, and gene-based profiling of antimicrobial resistance and virulence determinants. Results: All Italian isolates belonged to phylogroup B2 and to recognised ExPEC sequence types (ST706/O51:H1, ST141/O2:H6, ST372/O75:H31, ST646/O22:H5). Phenotypically, they were uniformly resistant to several penicillins and early/third-generation cephalosporins but remained susceptible to fluoroquinolones, aminoglycosides and trimethoprim–sulphonamide. The combined 57-genome pan-genome was open yet strongly core-dominated; Italian strains shared an efflux- and regulator-centred intrinsic resistome and a rich ExPEC virulence repertoire (P, S, F1C and type 1 fimbriae, multiple siderophores, colibactin, Vat, haemolysin, CNF1) with Brazilian and Finnish isolates. Conclusions: Pyometra-associated E. coli from northern Italian pets belong to globally disseminated high-risk B2 lineages that combine extensive virulence with a largely intrinsic resistome, and currently retain susceptibility to several key drug classes, underscoring an important but vulnerable therapeutic window.

Bacterial populations inhabiting a variety of natural and human-associated niches have the ability to grow in the form of biofilms. A large part of pathological chronic conditions, and essentially all the bacterial infections associated with implanted medical devices or prosthetics, are caused by microorganisms embedded in a matrix made of polysaccharides, proteins, and nucleic acids. Biofilm infections are generally characterized by a slow onset, mild symptoms, tendency to chronicity, and refractory response to antibiotic therapy. Even though the molecular mechanisms responsible for resistance to antimicrobial agents and host defenses have been deeply clarified, effective means to fight biofilms are still required. Lactic acid bacteria (LAB), used as probiotics, are emerging as powerful weapons to prevent adhesion, biofilm formation, and control overgrowth of pathogens. Hence, using probiotics or their metabolites to quench and interrupt bacterial communication and aggregation, and to interfere with biofilm formation and stability, might represent a new frontier in clinical microbiology and a valid alternative to antibiotic therapies. This review summarizes the current knowledge on the experimental and therapeutic applications of LAB to interfere with biofilm formation or disrupt the stability of pathogenic biofilms.

Background/Objectives: Staphylococcus aureus is a multifaceted pathogen responsible for diseases in humans and in several animal species, including dairy cows. This study aimed to characterize and compare the genetic diversity, lineage distribution, and antimicrobial resistance profiles of S. aureus isolates from bovine milk with human-derived reference genomes to investigate host adaptation and inter-species transmission. Methods: Genomic analyses were performed on S. aureus isolates from quarter milk samples of dairy cows together with human-derived sequences from public databases. Whole-genome sequencing and multi-locus sequence typing (MLST) were used to determine sequence type (ST) distribution, and the presence of key antibiotic resistance genes and mobile genetic elements (MGEs) was assessed. Comparative genomics was applied to evaluate gene content, phylogenetic relationships, and lineage–host associations. Results: The dataset encompassed bovine-adapted lineages (CC97, CC133, CC151) and human-associated lineages (CC1, CC5, CC8, CC30, CC45), as well as livestock-associated ST398 in bovine samples and human-adapted ST5 and ST6 in animals. ST8 was the only ST shared between animal and human isolates and showed differing resistance profiles, with animal ST8 carrying resistance determinants absent from human ST8. Bovine-adapted strains were characterized by recurrent loss of human-associated virulence genes and acquisition of bovine-associated mobile genetic elements, and blaZ and mecA were rarely detected in bovine-adapted CC97 but were frequently present in human CC5 and CC8. Overall, animal isolates carried fewer resistance genes than human isolates. Conclusions: S. aureus from dairy cows and humans displayed substantial genetic diversity, with evidence of host-associated lineages and dynamic changes in gene and mobile element content. These findings support the need for integrated One Health surveillance to track shared and host-adapted lineages and their antibiotic resistance determinants.

Background and Objectives: Biofilm formation on cochlear implants (CIs) poses a major problem for surgeons, leading to a high incidence of explantation and revision surgery. Therefore, developing appropriate and cost-effective biofilm detection and prevention techniques is of the essence. In this systematic review, we sought to investigate the development of biofilm formation on CIs. We also elaborated on experimental preventative biofilm measures. Materials and Methods: We conducted a systematic search of both in vitro and in vivo literature published in PubMed, Scopus, and ScienceDirect, until 15 June 2024, for published studies evaluating the biofilm formation and strategies for inhibiting biofilm formation on CIs. Depending on the type of the included study, we assessed quality with the modified Consolidated Standards of Reporting Trials tool, the Joanna Briggs Institute Case Reports Critical Appraisal Tool, a modified Delphi technique, and the ROBINS-I tool. We synthesized the available information on biofilm formation on CIs and the infection prevention capacity of the included antibiofilm agents. Results: A total of 26 studies were included in this systematic review. Biofilms in CIs are usually localized in their recesses such as their removable magnet pocket as opposed to their smooth surfaces. S. aureus and P. aeruginosa are the most commonly isolated microorganisms, and they tend to be strong biofilm producers. The optimal treatment strategy for a biofilm-infected CI is explantation. Most of the examined biofilm prevention methods in CIs present sufficient antibiofilm activity. Conclusions: Biofilm formation in CIs is considered one of the most dreadful complications. There have been no specific guidelines for the treatment of those cases, with removal and/or replacement of the CI being the treatment of choice. Various experimental prevention methods provide promising antibiofilm activity both in vivo and in vitro.

Purpose Periprosthetic joint infections induced by methicillin-resistant Staphylococcus aureus (MRSA) pose a major socioeconomic burden. Given the fact that MRSA carriers are at high risk for developing periprosthetic infections regardless of the administration of eradication treatment pre-operatively, the need for developing new prevention modalities is high. Methods The antibacterial and antibiofilm properties of vancomycin, Al 2 O 3 nanowires, and TiO 2 nanoparticles were evaluated in vitro using MIC and MBIC assays. MRSA biofilms were grown on titanium disks simulating orthopedic implants, and the infection prevention potential of vancomycin-, Al 2 O 3 nanowire-, and TiO 2 nanoparticle-supplemented Resomer® coating was evaluated against biofilm controls using the XTT reduction proliferation assay. Results Among the tested modalities, high- and low-dose vancomycin-loaded Resomer® coating yielded the most satisfactory metalwork protection against MRSA (median absorbance was 0.1705; [IQR = 0.1745] vs control absorbance 0.42 [IQR = 0.07]; p  = 0.016; biofilm reduction was 100%; and 0.209 [IQR = 0.1295] vs control 0.42 [IQR = 0.07]; p  < 0.001; biofilm reduction was 84%, respectively). On the other hand, polymer coating alone did not provide clinically meaningful biofilm growth prevention (median absorbance was 0.2585 [IQR = 0.1235] vs control 0.395 [IQR = 0.218]; p  < 0.001; biofilm reduction was 62%). Conclusions We advocate that apart from the well-established preventative measures for MRSA carriers, loading implants with bioresorbable Resomer® vancomycin-supplemented coating may decrease the incidence of early post-op surgical site infections with titanium implants. Of note, the payoff between localized toxicity and antibiofilm efficacy should be considered when loading polymers with highly concentrated antimicrobial agents.