Purpose: Imbalance in the microbiota, dysbiosis, has been identified in inflammatory bowel disease (IBD). We explored the fecal microbiota in pediatric patients with treatment-naïve IBD, non-IBD patients with gastrointestinal symptoms and healthy children, its relation to IBD subgroups, and treatment outcomes. Patients and methods: Fecal samples were collected from 235 children below 18 years of age. Eighty children had Crohn’s disease (CD), 27 ulcerative colitis (UC), 3 IBD unclassified, 50 were non-IBD symptomatic patients, and 75 were healthy. The bacterial abundance of 54 predefined DNA markers was measured with a 16S rRNA DNA-based test using GA-Map™ technology at diagnosis and after therapy in IBD patients. Results: Bacterial abundance was similarly reduced in IBD and non-IBD patients in 51 of 54 markers compared to healthy patients (P<0.001). Only Prevotella was more abundant in patients (P<0.01). IBD patients with ileocolitis or total colitis had more Ruminococcus gnavus (P=0.02) than patients with colonic CD or left-sided UC. CD patients with upper gastrointestinal manifestations had higher Veillonella abundance (P<0.01). IBD patients (58%) who received biologic therapy had lower baseline Firmicutes and Mycoplasma hominis abundance (P<0.01) than conventionally treated. High Proteobacteria abundance was associated with stricturing/penetrating CD, surgery (P<0.01), and nonmucosal healing (P<0.03). Low Faecalibacterium prausnitzii abundance was associated with prior antibiotic therapy (P=0.001), surgery (P=0.02), and nonmucosal healing (P<0.03). After therapy, IBD patients had unchanged dysbiosis. Conclusion: Fecal microbiota profiles differentiated IBD and non-IBD symptomatic children from healthy children, but displayed similar dysbiosis in IBD and non-IBD symptomatic patients. Pretreatment fecal microbiota profiles may be of prognostic value and aid in treatment individualization in pediatric IBD as severe dysbiosis was associated with an extensive, complicated phenotype, biologic therapy, and nonmucosal healing. The dysbiosis persisted after therapy, regardless of treatments and mucosal healing.

Background: Staphylococcus aureus is the main microbial pathogen in orthopaedic infections, and it adds considerable extra costs to the national health-care system each year. Nasal carriers of Staphylococcus aureus have an increased risk of invasive disease, including surgical site infection. The purpose of the present study was to investigate whether the Staphylococcus aureus carrier clones found in patients undergoing elective orthopaedic surgery were the same as the clones found in isolates from orthopaedic patients with Staphylococcus aureus surgical site infections. Methods: Patients admitted for elective orthopaedic surgery underwent nasal cultures for Staphylococcus aureus . Further, orthopaedic patients with a deep surgical site infection caused by Staphylococcus aureus were characterized using the same genotyping methods: multilocus sequence typing and staphylococcal protein A typing. Results: Multilocus sequence typing revealed a large number of genotypes in the two populations. However, 85% of nasal carriers and 90% of surgical site infection isolates could be classified into the same four multilocus sequence typing clonal complexes. The risk of Staphylococcus aureus surgical site infection in nasal carriers compared with non-carriers was 5.8 times higher (95% confidence interval, 1.5 to 23.1 times). Of the nasal carriers, 6.3% (95% confidence interval, 1.7% to 10.9% [seven of 111 patients]) developed a deep Staphylococcus aureus surgical site infection, and all but one patient had identical genotypes in the nasal and surgical site infection isolates. Conclusions: Staphylococcus aureus isolates from nasal carriers and patients with surgical site infection clustered into the same few multilocus sequence typing clonal complexes. This finding confirms the existence of some commonly occurring Staphylococcus aureus clones in different population groups within a geographically restricted area. The almost complete individual concordance between Staphylococcus aureus genotypes in carriers who developed a deep surgical site infection strongly supports transmission from the nose, skin surfaces, and other endogenous body regions as a possible route. Clinical Relevance: Surgical site infections might be more frequently caused by endogenous transmission than was previously assumed. Perioperative preventive efforts must focus more on this route to further decrease the risk of postoperative orthopaedic infections.

Significance: Methicillin-resistant Staphylococcus aureus (MRSA), traditionally associated with hospitals, is increasingly circulating in the community. This imposes, in turn, a potential burden on hospital infection control due to a more frequent hospitalization of colonized patients. We developed an individual-based model, reproducing community and healthcare settings, to understand the epidemiological drivers of MRSA and the connections between the society and the healthcare institutions. We show that in Norway, a low-prevalence country, the rise of infections is driven by an increasing inflow of cases from abroad rather than by an ongoing epidemic. We demonstrate the major role played by households in transmitting MRSA and show that the burden on hospitals from the growing community circulation is still limited thanks to aggressive infection-control protocols.

Understanding the evolution of CA-MRSA is important in light of the increasing importance of this reservoir in the dissemination of MRSA. Here, we highlight the story of the CA-MRSA CC152 lineage using whole-genome sequencing on an international collection of CC152. We show that the evolution of this lineage is novel and that antibiotic usage may have the potential to select for the phage-encoded Panton-Valentine leukocidin. The diversity of the strains correlated highly to geography, with higher level of resistance observed among the European MRSA isolates. The mobility of the SCC mec element is mandatory for the emergence of novel MRSA lineages, and we show here distinct acquisitions, one of which is linked to the successful clone found throughout Europe today.

Nucleic acid purification methods are of importance when performing microbiota studies and especially when analysing the intestinal microbiota as we here find a wide range of different microbes. Various considerations must be taken to lyse the microbial cell wall of each microbe. In the present article, we compare several tissue lysis steps and commercial purification kits, to achieve a joint RNA and DNA purification protocol for the purpose of investigating the microbiota and the microbiota-host interactions in a single colonic mucosal tissue sample. A further optimised tissue homogenisation and lysis protocol comprising mechanical bead beating, lysis buffer replacement and enzymatic treatment, in combination with the AllPrep DNA/RNA Mini Kit (Qiagen, Hilden, Germany) resulted in efficient and simultaneous purification of microbial and human RNA and DNA from a single mucosal colonic tissue sample. The present work provides a unique possibility to study RNA and DNA from the same mucosal biopsy sample, making a direct comparison between metabolically active microbes and total microbial DNA. The protocol also offers an opportunity to investigate other members of a microbiota such as viruses, fungi and micro-eukaryotes, and moreover the possibility to extract data on microbiota and host interactions from one single mucosal biopsy.

Colorectal epithelial tumors occur spontaneously in dogs, and the pathogenesis seems to parallel that of humans. The development of human colorectal tumorigenesis has been linked to alterations in the composition of the intestinal microbiota. This study characterized the fecal- and mucosa-associated microbiota in dogs with colorectal epithelial tumors (n = 10). The fecal microbiota was characterized by 16S rDNA analysis and compared with that of control dogs (n = 13). We also determined the mucosa-associated microbiota composition in colonic tumor tissue (n = 8) and in adjacent non-tumor tissue (n = 5) by 16S rDNA- and rRNA profiling. The fecal microbial community structure in dogs with tumors was different from that of control samples and was distinguished by oligotypes affiliated with Enterobacteriaceae, Bacteroides, Helicobacter, Porphyromonas, Peptostreptococcus and Streptococcus, and lower abundance of Ruminococcaceae, Slackia, Clostridium XI and Faecalibacterium. The overall community structure and populations of mucosal bacteria were not different based on either the 16S rDNA or the 16S rRNA profile in tumor tissue vs. adjacent non-tumor tissue. However, the proportion of live, potentially active bacteria appeared to be higher in non-tumor tissue compared with tumor tissue and included Slackia, Roseburia, unclass. Ruminococcaeceae, unclass. Lachnospiraceae and Oscillibacter. Colorectal tumors are rarely diagnosed in dogs, but despite this limitation, we were able to show that dogs with colorectal tumors have distinct fecal microbiota profiles. These initial results support the need for future case-control studies that are adequately powered, as well as age-matched and breed-matched, in order to evaluate the influence of bacteria on colorectal cancer etiopathogenesis and to determine whether the bacteria may have potential as biomarkers in clinical settings.