Genomic analysis of variability in Delta-toxin levels between Staphylococcus aureus strains.

BACKGROUND: The delta-toxin (δ-toxin) of Staphylococcus aureus is the only hemolysin shown to cause mast cell degranulation and is linked to atopic dermatitis, a chronic inflammatory skin disease. We sought to characterize variation in δ-toxin production across S. aureus strains and identify genetic loci potentially associated with differences between strains. METHODS: A set of 124 S. aureus strains was genome-sequenced and δ-toxin levels in stationary phase supernatants determined by high performance liquid chromatography (HPLC). SNPs and kmers were associated with differences in toxin production using four genome-wide association study (GWAS) methods. Transposon mutations in candidate genes were tested for their δ-toxin levels. We constructed XGBoost models to predict toxin production based on genetic loci discovered to be potentially associated with the phenotype. RESULTS: The S. aureus strain set encompassed 40 sequence types (STs) in 23 clonal complexes (CCs). δ-toxin production ranged from barely detectable levels to >90,000 units, with a median of >8,000 units. CC30 had significantly lower levels of toxin production than average while CC45 and CC121 were higher. MSSA (methicillin sensitive) strains had higher δ-toxin production than MRSA (methicillin resistant) strains. Through multiple GWAS approaches, 45 genes were found to be potentially associated with toxicity. Machine learning models using loci discovered through GWAS as features were able to predict δ-toxin production (as a high/low binary phenotype) with a precision of .875 and specificity of .990 but recall of .333. We discovered that mutants in the carA gene, encoding the small chain of carbamoyl phosphate synthase, completely abolished toxin production and toxicity in Caenorhabditis elegans. CONCLUSIONS: The amount of stationary phase production of the toxin is a strain-specific phenotype likely affected by a complex interaction of number of genes with different levels of effect. We discovered new candidate genes that potentially play a role in modulating production. We report for the first time that the product of the carA gene is necessary for δ-toxin production in USA300. This work lays a foundation for future work on understanding toxin regulation in S. aureus and prediction of phenotypes from genomic sequences.

A Comparison of the Bacterial Nasal Microbiome in Allergic Rhinitis Patients Before and After Immunotherapy.

OBJECTIVES/HYPOTHESIS: Recent research has examined the nasal microbiome in rhinosinusitis and nondiseased states. Given immunologic alterations in allergic rhinitis (AR) and after allergen immunotherapy (IT), we evaluated the nasal microbiome in these conditions. STUDY DESIGN: Cross-sectional comparison. METHODS: In this cross-sectional study, nasal swabs for microbiome analysis were collected from three patient groups: IT-naïve AR patients, AR patients undergoing IT for greater than 12 months, and a control group without sinonasal inflammatory disease. RESULTS: Nasal swabs were successfully collected for 14 IT-naïve AR patients, 20 post-IT patients, and 17 controls. The α diversity showed a statistical difference in evenness but not in richness amongst samples, whereas the ß-diversity was significantly different between groups. Corynebacterium and Staphylococcus were the most prevalent bacteria across all groups. CONCLUSIONS: ß-diversity was found to be significantly different across the three groups, but the AR groups were found to be more similar to each other than to the controls. Although there is symptomatic improvement in the AR group undergoing IT, the microbiome does not appear to transition to a healthy microbiome composition. LEVEL OF EVIDENCE: 4 Laryngoscope, 2020.

High Rates of New Delhi Metallo-ß-Lactamase Carbapenemase Genes in Multi-Drug Resistant Gram-Negative Bacteria in Nicaragua.

Antimicrobial resistance (AMR) is a global public health crisis. Much of the burden of AMR in resource-limited settings remains unknown. This pilot study characterized clinical isolates of multidrug-resistant Gram-negative rods (MDR-GNRs) from Nicaragua. New Delhi metallo-ß-lactamase (NDM) carbapenemase genes were detected in 60% of isolates. Enterobacteriaceae had the highest rates of NDM detection, with 92% (50/54 isolates) positive by polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) analysis revealed patterns of clustering among isolates by two factors: plasmid profiles and year of culture. These findings of very high rates of NDM-carbapenemase genes in MDR-GNRs from hospitals throughout Nicaragua are alarming. Further research is needed to determine clinical and epidemiologic factors associated with multidrug-resistant isolates and to guide interventions to limit further spread.