Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is an important emerging zoonotic pathogen that causes severe skin infections. To combat infections from drug-resistant bacteria, the transplantation of commensal antimicrobial bacteria as a therapeutic has shown clinical promise. We screened a collection of diverse staphylococcus species from domestic dogs and cats for antimicrobial activity against MRSP. A unique strain (S. felis C4) was isolated from feline skin that inhibited MRSP and multiple gram-positive pathogens. Competition experiments in mice showed that S. felis significantly reduced MRSP skin colonization and an antimicrobial extract from S. felis significantly reduced necrotic skin injury from MRSP infection. Fluorescence and electron microscopy revealed that S. felis antimicrobials disrupted bacterial but not eukaryotic cell membranes. LC/MS identified several S. felis phenol-soluble modulin beta (PSMβ) peptides that exhibited antimicrobial and anti-inflammatory activity. These findings indicate a feline commensal bacterium that could be utilized in bacteriotherapy against difficult-to-treat animal and human skin infections.

NHEK cells were treated with DMSO (0.1%) control, PSMβ2 (10 μg/ml), Poly I:C (0.4 μg/ml) or PSMβ2 and Poly I:C combined, all in triplicate wells, for 4 h or 24 h in and RNA was extracted using the PureLink RNA mini kit and triplicate samples were pooled together for each treatment. Isolated RNA was submitted to the UCSD IGM Genomics Center for RNA-sequencing performed on a high-output run V4 platform (Illumina, USA) with a single read 100 cycle runs. All the steps including trim bases, alignment with bowtie 2, quantification to annotation model (Partek E.M), Normalization (CPM/Add 1.0 E-4), Gene Set Enrichment (GSA), Differential Gene Analysis and hierarchial clustering, and Gene Ontology analysis was performed with the Partek Flow Genomics Suite.

contributor: Alan O'Neill                            
contributor: Nikhil Kulkarni                            
                                
SAMPLES
Sample name    source name                          organism                                                    characteristics: treatment                                   characteristics: time            
Sample 1          Control_4_h_S76                  NHEK    Human primary keratinocytes    DMSO (0.1%)                                                   4 h post-treatment            
Sample 2          Control_24_h_S80                NHEK    Human primary keratinocytes    DMSO (0.1%)                                                   24 h post-treatment            
Sample 3          Poly_IC_4_h_S78                 NHEK    Human primary keratinocytes    Poly I:C (0.4 ug/ml)                                            4 h post-treatment            
Sample 4          Poly_IC_24_h_S82               NHEK    Human primary keratinocytes    Poly I:C (0.4 ug/ml)                                          24 h post-treatment            
Sample 5          Poly_IC_PSMB_4_h_S79    NHEK    Human primary keratinocytes    Poly I:C (0.4 ug/ml) and PSMB2 (10 ug/ml)    4 h post-treatment            
Sample 6          Poly_IC_PSMB_24_h_S83  NHEK    Human primary keratinocytes    Poly I:C (0.4 ug/ml) and PSMB2 (10 ug/ml)    24 h post-treatment            
Sample 7          PSMB_4_h_S77                    NHEK    Human primary keratinocytes     PSM?2 (10 ug/ml)                                             4 h post-treatment            
Sample 8          PSMB_24_h_S81                 NHEK    Human primary keratinocytes     PSM?2 (10 ug/ml)                                             24 h post-treatment            
                                
PROTOCOLS
growth protocol: NHEK cells were cultured in EpiLife medium supplemented with EpiLife Defined Growth Supplement (EDGS)
treatment protocol: "NHEK cells were treated with DMSO (0.1%), PolyI:C (0.4 ?g/ml), PSM?2 (10 ?g/ml) or PolyI:C and PSM?2 for 4 h or 24 h."                            
extract protocol: NHEKs cells were washed 2X with PBS and RNA was extracted using PureLink RNA Mini Kit
                                
DATA PROCESSING PIPELINE
data processing step: "All the steps including trim bases, alignment with bowtie 2, quantification to annotation model (Partek E.M), Normalization (CPM/Add 1.0 E-4), Gene Set Enrichment (GSA), Differential Gene Analysis and hierarchial clustering, and Gene Ontology analysis was performed with the Partek Flow Genomics Suite."                       
genome build: hg19                            
processed data files format and content: txt