Commensal Escherichia coli, naturally occurring in the intestinal tract, can be the origin of extraintestinal pathogenic E. coli (ExPEC) strains. ExPEC causes high mortality and significant economic losses in the swine industry in several countries and poses a serious threat to public health worldwide. The aim of this study was to analyze the extended phylogenetic structure and extraintestinal virulence potential in two groups of commensal E. coli isolates from post-weaning piglets and sows. The phylogenetic assignment to eight groups was determined using the revised Clermont phylogenetic typing method in quadruplex PCR. Identification of extraintestinal virulence genes (VGs) and adhesin operon genes was performed using multiplex or simplex PCR. The revised phylogenetic assignment allowed us to distinguish E. coli with significantly higher (groups C and F) or lower (group E) virulence potential in isolates from piglets. The majority of the tested VGs occurred more frequently in isolates from piglets than from sows, with statistically significant differences for seven genes: fimH, papAH, iutA, iroN, ompT, traT, and iss. Complete operons for type I and P fimbriae significantly prevailed among E. coli from piglets. This study provides insight into the extended phylogenetic structure of porcine commensal E. coli and showed that these strains, particularly from piglets, constitute a considerable reservoir of extraintestinal VGs and may increase the potential risk of extraintestinal infections.
Background: In the oral and maxillofacial surgery, fixation plates are commonly used for the stabilization of bone fragments. Additive manufacturing has enabled us to design and create personalized fixation devices that would ideally fit any given fracture. Aim: The aim of the present preliminary study was to assess the susceptibility of 3D-printed titanium fixation plates to biofilm formation. Methods: Plates were manufactured using selective laser melting (SLM) from Ti-6Al-4 V. Reference strains of Streptococcus mutans, Staphyloccocus epidermidis, Staphylococcus aureus, Lactobacillus rhamnosus, and Candida albicans, were tested to evaluate the material’s susceptibility to biofilm formation over 48 hours. Biofilm formations were quantified by a colorimetric method and colony-forming units (CFU) quantification. Scanning electron microscopy (SEM) visualized the structure of the biofilm. Results: Surface analysis revealed the average roughness of 102.75 nm and irregular topography of the tested plates. They were susceptible to biofilm formation by all tested strains. The average CFUs were as follows: S. mutans (11.91 x 107) > S.epidermidis (4.45 x 107) > S. aureus (2.3 x 107) > C.albicans (1.22 x 107) > L. rhamnosus (0.78 x 107). Conclusions: The present preliminary study showed that rough surfaces of additively manufactured titanium plates are susceptible to microbial adhesion. The research should be continued in order to compare additively manufactured plates with other commercially available osteotomy plates. Therefore, we suggest caution when using this type of material.
The regulatory functions of microRNA (miRNA) are involved in all processes contributing to carcinogenesis and response to viral infections. Cervical cancer in most cases is caused by the persistence of high-risk human papillomavirus (HR-HPV) infection. While oncogenic human papillomaviruses induce aberrant expression of many cellular miRNAs, this dysregulation could be harnessed as a marker in early diagnosis of HR-HPV infection, cervical squamous intraepithelial lesions, and cancer. In recent years, growing data indicate that miRNAs show specific patterns at various stages of cervical pathology. The aim of this review is to systematize current reports on miRNA capacity that can be utilized in personalized diagnostics of cervical precancerous and cancerous lesions. The analysis of the resources available in online databases (National Center for Biotechnology Information—NCBI, PubMed, ScienceDirect, Scopus) was performed. To date, no standardized diagnostic algorithm using the miRNA pattern in cervical pathology has been defined. However, the high sensitivity and specificity of the reported assays gives hope for the development of non-invasive diagnostic tests that take into account the heterogeneity of tumor-related changes. Due to this variability resulting in difficult to predict clinical outcomes, precise molecular tools are needed to improve the diagnostic and therapeutic process.
New hydrogel materials developed to improve soft tissue healing are an alternative for medical applications, such as tissue regeneration or enhancing the biotolerance effect in the tissue-implant–body fluid system. The biggest advantages of hydrogel materials are the presence of a large amount of water and a polymeric structure that corresponds to the extracellular matrix, which allows to create healing conditions similar to physiological ones. The present work deals with the change in mechanical properties of sodium alginate mixed with gelatin containing Pygeum africanum. The work primarily concentrates on the evaluation of the mechanical properties of the hydrogel materials produced by the sol–gel method. The antimicrobial activity of the hydrogels was investigated based on the population growth dynamics of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923, as well as the degree of degradation after contact with urine using an innovative method with a urine flow simulation stand. On the basis of mechanical tests, it was found that sodium alginate-based hydrogels with gelatin showed weaker mechanical properties than without the additive. In addition, gelatin accelerates the degradation process of the produced hydrogel materials. Antimicrobial studies have shown that the presence of African plum bark extract in the hydrogel enhances the inhibitory effect on Gram-positive and Gram-negative bacteria. The research topic was considered due to the increased demand from patients for medical devices to promote healing of urethral epithelial injuries in order to prevent the formation of urethral strictures.
Antibiotic resistance concerns various areas with high consumption of antibiotics, including husbandry. Resistant strains are transmitted to humans from livestock and agricultural products via the food chain and may pose a health risk. The commensal microbiota protects against the invasion of environmental strains by secretion of bacteriocins, among other mechanisms. The present study aims to characterize the bactericidal potential of bacteriocinogenic Escherichia coli from healthy humans against multidrug-resistant and antibiotic-sensitive strains from pigs and cattle. Bacteriocin production was tested by the double-layer plate method, and bacteriocin genes were identified by the PCR method. At least one bacteriocinogenic E. coli was detected in the fecal samples of 55% of tested individuals, adults and children. Among all isolates (n = 210), 37.1% were bacteriocinogenic and contained genes of colicin (Col) Ib, ColE1, microcin (Mcc) H47, ColIa, ColM, MccV, ColK, ColB, and single ColE2 and ColE7. Twenty-five E. coli carrying various sets of bacteriocin genes were further characterized and tested for their activity against zoonotic strains (n = 60). Strains with ColE7 (88%), ColE1-ColIa-ColK-MccH47 (85%), MccH47-MccV (85%), ColE1-ColIa-ColM (82%), ColE1 (75%), ColM (67%), and ColK (65%) were most active against zoonotic strains. Statistically significant differences in activity toward antibiotic-resistant strains were shown by commensal E. coli carrying MccV, ColK-MccV, and ColIb-ColK. The study demonstrates that bacteriocinogenic commensal E. coli exerts antagonistic activity against zoonotic strains and may constitute a defense line against multidrug-resistant strains.
Background: The commensal Escherichia coli population may significantly influence the pathogenesis of extraintestinal infections. The assignment to specific sequence type (ST) lineages and the presence of particular combinations of virulence genes are characteristic features of extraintestinal pathogenic E. coli (ExPEC)—although not exclusively. Extraintestinal virulence genes are also identified among commensal E. coli. This study aimed to examine the genotypic and phenotypic characteristics of the extraintestinal virulence potential of two populations of commensal E. coli isolates from adults and young children.Methods: Genotypic traits were detected using polymerase chain reaction (PCR). Appropriate phenotypic assays and real-time PCR were used to analyze the expression of virulence factors. Multilocus sequence typing was performed using the seven-loci Achtman scheme.Results: Genotypic studies revealed the virulence potential of the commensal isolates, and phenotypic analyses confirmed whether the genes are expressed. E. coli from adults carried pathogenicity islands and virulence genes in significantly higher proportions, resulting from the dominance of phylogroup B2 in this set of isolates. The hemolytic activity and higher levels of siderophore receptor expression were more common in E. coli from adults and were closely related to the dominance of phylogroup B2. Other traits not associated with phylogroup B2, such as adhesion abilities mediated by type 1 and P fimbriae and strong biofilm formation propensities, were detected with similar frequencies in both pools of isolates. E. coli harboring pathogenicity islands were subjected to multilocus sequence typing analysis. Sequence types ST73, ST59, ST131, ST95, ST141, and ST69 were most common in isolates from adults, whereas ST10, ST155, ST59, and ST1823 from children. In our collection of E. coli, the ST73 exhibited the highest potential for extraintestinal virulence.Conclusion: A significant proportion of E. coli from adults compared to young children exhibited considerable virulence potential, which may enable them to function as endogenous pathogens. Our research highlights the significant accumulation of extraintestinal pathogenicity features in commensal E. coli, indicating the need to monitor genetic and phenotypic profiles in “silent” potential pathogens.