Proteus mirabilis is one of the most common etiological agent of complicated urinary tract infections, specially those associated to catheterization. This is related to the ability of P. mirabilis to form biofilms on different surfaces. This pathogen encodes 17 putative fimbrial operons, the highest amount found in any sequenced bacterial species so far. The present study analyzed the role of 4 P. mirabilis fimbriae (MR/P, UCA, ATF and PMF) in biofilm formation using isogenic mutants. Experimental approaches included migration over cathether, swimming and swarming motility, the semiquantitative assay based on adhesion and crystal violet staining and biofilm development by immunofluorescence and confocal microscopy. Different assays were performed using LB or artificial urine. Results indicated that the different fimbriae contribute to the formation of a stable and functional biofilm. Fimbriae revealed particular associated roles. First, all the mutants showed a significantly reduced ability to migrate across urinary catheters sections but neither swimming nor swarming motility were affected. However some mutants formed smaller biofilms compared with the wild type (MRP and ATF) while others formed significantly larger biofilms (UCA and PMF) showing different bioarchitecture features. It can be concluded that P. mirabilis fimbriae have distinguishable roles in the generation of biofilms, particulary associated to catheters.

Recurrent urinary tract infections (UTIs) occur frequently in children and women. Intracellular bacterial communities (IBCs) and biofilm formation by Escherichia coli are risk factors for recurrence. The aim of this study was to evaluate the effect of different antibiotics on biofilms by E. coli strains isolated from children with UTI and to correlate virulence factors and IBCs with biofilm formation. A total of 116 E. coli strains were tested for biofilm formation using the crystal violet microplate technique. 58.6% of the strains did not produce biofilm, while 16.4%, 18.1% and 6.8% formed weak, moderate and strong biofilms, respectively. No correlation was found between the ability to form biofilms and the presence of IBCs. Biofilm formation was significantly associated with pili P codifying genes, whereas other virulence factors were not statistically associated. Antibiotics, including ampicillin, cephalothin, ceftriaxone, ceftazidime, amikacin and ciprofloxacin, were evaluated at different concentrations after 48 h of biofilm formation. Except ampicillin, the other antibiotics tested induced a significant reduction of biofilm biomass. In the case of recurrent UTIs potentially associated with the presence of biofilm, the use of third-generation cephalosporin, fluoroquinolones and aminoglycosides could be recommended. These antibiotics demonstrated to reduce biofilm biomass produced even by resistant strains.

Aim: Proteus mirabilis biofilms colonize medical devices, and their role in microbial pathogenesis is well established. Magnesium-doped zinc oxide nanoparticles (ZnO:MgO NPs) have potential antimic...

Proteus mirabilis(P. mirabilis) is a common etiological agent of urinary tract infections, particularly those associated with catheterization. P. mirabilis efficiently forms biofilms on different surfaces and shows a multicellular behavior called 'swarming', mediated by flagella. To date, the role of flagella in P. mirabilis biofilm formation has been under debate. In this study, we assessed the role of P. mirabilis flagella in biofilm formation using an isogenic allelic replacement mutant unable to express flagellin. Different approaches were used, such as the evaluation of cell surface hydrophobicity, bacterial motility and migration across catheter sections, measurements of biofilm biomass and biofilm dynamics by immunofluorescence and confocal microscopy in static and flow models. Our findings indicate that P. mirabilis flagella play a role in biofilm formation, although their lack does not completely avoid biofilm generation. Our data suggest that impairment of flagellar function can contribute to biofilm prevention in the context of strategies focused on particular bacterial targets.

Acinetobacter baumannii is a relevant opportunistic pathogen, and one of the main microorganisms responsible for outbreaks in nosocomial infections worldwide. Its pathogenicity is mainly due to its resistance to multiple antibiotics and to its ability to form biofilms on abiotic surfaces. The objective of this study was to characterize the biofilm formation cycle of A. baumannii isolated from a patient in a hospital and compare its antibiotic resistance with the planktonic cells. To study biofilm formation, the classical microtiter assay was used, with crystal violet staining and optical density reading to classify the type of biofilm. Also, the effect of gentamicin and colistin on bacterial biofilm was studied with an extra step of antibiotic addition. For the characterization of the different biofilm formation stages, the strain was grown on a coverslip, and the stain was made with a mixture of fluorophores markers to visualize the biofilm with a confocal laser microscope. It was possible to differentiate the A. baumannii biofilm formation stages. Through these observations, it was possible to estimate the time elapsed between each stage. As the strain was susceptible to colistin and gentamicin, both antibiotics were evaluated after the biofilm was formed. Neither antibiotics showed an effect on the eradication of A. baumannii biofilm.

Canine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (ddPCR) in comparison to conventional reverse-transcription (RT-PCR) and quantitative reverse-transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from apparently healthy individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR exhibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR showed a moderate concordance, with a Kappa coefficient of 0.477. The sensitivity was 42.4% for RT-PCR, 57.9% for RT-qPCR, and 72.4% for ddPCR, with 100% specificity for all methods. This study underscores ddPCR’s superior sensitivity and agreement with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis.