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Chronic diabetic foot is a global burden affecting millions of people, and the chronicity of an ulcer is directly linked to the diverse bacterial burden and its biofilm mode of infection. The bacterial diversity of 100 chronic diabetic ulcer samples was profiled via traditional culturing method as well as metagenomic approach by sequencing the 16S rRNA V3 hyper-variable region on Illumina Miseq Platform (Illumina, Inc., San Diego, CA). All the relevant clinical metadata, including duration of diabetes, grade of ulcer, presence of neuropathy, and glycaemic level, were noted and correlated with the microbiota. The occurrence and establishment of bacterial biofilm over chronic wound tissues was revealed by Fluorescent in situ Hybridization and Scanning Electron Microscopy. The biofilm-forming ability of predominant bacterial isolates was studied via crystal violet assay and Confocal Laser Scanning Microscopy. The dominant phyla obtained from bacterial diversity analysis were Firmicutes, Proteobacteria, and Actinobacteria. The dominant aerobic pathogens identified by culture method are Pseudomonas, Proteus, Enterococcus, and Staphylococcus, whereas high-throughput sequencing revealed heightened levels of Streptococcus and Corynebacterium along with 22 different obligate anaerobes. The biofilm occurrence in chronic diabetic ulcer infection is well analysed. Herein, we illustrate the comprehensive pattern of bacterial infection and identify the community composition of chronic wound pathogenic biofilm.

Multidrug-resistant Staphylococcus aureus is a leading concern worldwide. Coagulase-Negative Staphylococci are claimed to be the reservoir and source of important resistant elements in S. aureus. However, the origin and evolutionary route of resistant genes in S. aureus are still remaining unknown. Here, we performed a detailed phylogenomic analysis of 152 completely sequenced S. aureus strains in comparison with 7,529 non-Staphylococcus aureus reference bacterial genomes. Our results reveal that S. aureus has a large open pan-genome where 97 (55%) of its known resistant-related genes belonging to its accessory genome. Among these genes, 47 (27%) were located within the Staphylococcal Cassette Chromosome mec (SCCmec), a transposable element responsible for resistance against major classes of antibiotics including beta-lactams, macrolides, and aminoglycosides. However, the physically linked mec-box genes (MecA–MecR–MecI) that are responsible for the maintenance of SCCmec elements is not unique to S. aureus, instead it is widely distributed within Staphylococcaceae family. The phyletic patterns of SCCmec-encoded resistant genes in Staphylococcus species are significantly different from that of its core genes indicating frequent exchange of these genes between Staphylococcus species. Our in-depth analysis of SCCmec-resistant gene phylogenies reveals that genes such as blaZ, ble, kmA, and tetK that are responsible for beta-lactam, bleomycin, kanamycin, and tetracycline resistance in S. aureus were laterally transferred from non-Staphylococcus sources. In addition, at least 11 non-SCCmec-encoded resistant genes in S. aureus, were laterally acquired from distantly related species. Our study evidently shows that gene transfers played a crucial role in shaping the evolution of antibiotic resistance in S. aureus.

Wohlfahrtiimonas chitiniclastica is a rare pathogen that was first isolated from a parasitic fly, Wohlfahrtia magnifica. It is a very rare but an emerging human pathogen reported only in Europe and South America. Recently, it is reported to be an egressing zoonotic pathogen from different geographical locations. The present case represents the first report of this pathogen from a patient suffering from osteomyelitis from India and so far no reports are available regarding the W. chitiniclastica associated infections in Asian countries. Clinical awareness of such emerging human pathogens is crucial for the infectious disease containment.

We report the outcome of 94 Miller-Galante unicompartmental knee replacements done by a single surgeon over 16 years in a district general hospital in the UK. Patellofemoral arthritis was considered a contraindication only if symptomatic and confirmed on radiography and surgery. Deformity when present was correctable. The mean age at surgery was 66.54 years. The mean followup was 10.8 years (2-16 years). The mean Bristol knee score was 43.1 (28-50); 86% of the patients had good or excellent scores. The average range of flexion was 110.6°( 80-130°); 89% of the knees had an appropriate alignment. Using revision as the endpoint, outcome for every knee was established. The survival rate for medial unicompartmental knee replacements was 94% at ten years and 87% at 15 years. Although the survival rate for the lateral unicompartmental knee replacement was 97% at five years, it dropped to 41% at eight years. Lateral unicompartmental knee replacements formed only 9.6% of all the replacements in this series. The results for medial unicompartmental knee replacements are similar to reports by other authors for similar and mobile bearing designs. Unicompartmental knee replacement results in a more kinematic knee and produces good functional results.

Background Enterococcus faecalis is a major clinically relevant nosocomial bacterial pathogen frequently isolated from polymicrobial infections. The biofilm forming ability of E. faecalis attributes a key role in its virulence and drug resistance. Biofilm cells are phenotypically and metabolically different from their planktonic counterparts and many aspects involved in E. faecalis biofilm formation are yet to be elucidated. The strain E. faecalis SK460 used in the present study is esp (Enterococcal surface protein) and fsr (two-component signal transduction system) negative non-gelatinase producing strong biofilm former isolated from a chronic diabetic foot ulcer patient. We executed a label-free quantitative proteomic approach to elucidate the differential protein expression pattern at planktonic and biofilm stages of SK460 to come up with potential determinants associated with Enterococcal biofilm formation. Results The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of proteomic data revealed that biofilm cells expressed higher levels of proteins which are associated with glycolysis, amino acid biosynthesis, biosynthesis of secondary metabolites, microbial metabolism in diverse environments and stress response factors. Besides these basic survival pathways, LuxS-mediated quorum sensing, arginine metabolism, rhamnose biosynthesis, pheromone and adhesion associated proteins were found to be upregulated during the biofilm transit from planktonic stages. The selected subsets were validated by quantitative real-time PCR. In silico functional interaction analysis revealed that the genes involved in upregulated pathways pose a close molecular interaction thereby coordinating the regulatory network to thrive as a biofilm community. Conclusions The present study describes the first report of the quantitative proteome analysis of an esp and fsr negative non gelatinase producing E. faecalis . Proteome analysis evidenced enhanced expression of glycolytic pathways, stress response factors, LuxS quorum signaling system, rhamnopolysaccharide synthesis and pheromone associated proteins in biofilm phenotype. We also pointed out the relevance of LuxS quorum sensing and pheromone associated proteins in the biofilm development of E. faecalis which lacks the Fsr quorum signaling system. These validated biofilm determinants can act as potential inhibiting targets in Enterococcal infections. Electronic supplementary material The online version of this article (10.1186/s12866-019-1527-2) contains supplementary material, which is available to authorized users.

Anteroposterior radiographs of the fully flexed and slightly externally rotated elbow were obtained in 122 cases of cubital tunnel syndrome (CUTS) and in 33 normal elbows. Medial trochlear lip osteophytes were not found in the studies of normal elbows but were noted in 20% of ulnar nerve entrapment cases. Medial incongruity between the trochlea and the olecranon greater than or equal to 5 mm was found in 29% of CUTS cases and in 6% of normal elbows. The presence of posttraumatic elbow-joint deformities seen on radiographs correlated significantly with the degree of clinical involvement. Depth and area measurements of ulnar nerve compression, performed using a line-art representation of the radiographs, showed a significant (P less than .0001) correlation between decrease in the area of the cubital tunnel and ulnar innervated muscle atrophy. These findings confirm the importance in this syndrome of both osteoarthritis and incongruity of the medial structures of the elbow joint.