title: Capsular hyaluronic acid of equine isolates of Streptococcus zooepidemicus is upregulated at temperatures below 35°C.
abstract: BACKGROUND
  Streptococcus zooepidemicus causes opportunist respiratory and other infections in the horse. Capsule expression is highly variable and known to affect resistance to phagocytosis. Most clinical isolates producing small, dry colonies at 37°C produce mucoid colonies at temperatures below 35°C.
  
  
  OBJECTIVE
  The aim was to understand the molecular basis of increased capsule expression by equine isolates of S. zooepidemicus at temperatures lower than 35°C.
  
  
  METHODS
  Cross-sectional observational study.
  
  
  METHODS
  Capsule production by groups of equine S. zooepidemicus strains was determined at 23, 30, 35 and 37°C. Hyaluronidase (HylC) at 23 and 37°C was measured by quantitative enzyme-linked immunosorbent assay. Expressions of hasA and hylC at 23 and 37°C were measured by quantitative reverse transcriptase-PCR. The covRS genes in representative S. zooepidemicus were sequenced and checked for mutations.
  
  
  RESULTS
  Colonies of randomly selected S. zooepidemicus strains became mucoid or showed marked increase in colony mucoidy following the change in temperature to 23°C. Expression of hasA at 23°C was 45- to 700-fold greater than at 37°C. Transcription of hylC at 23°C was 2.5- to 200-fold greater than at 37°C, yet enzyme concentrations in cultures were significantly higher at 37°C (P<0.05), suggesting that production of HylC is regulated post transcriptionally. The covRS gene in S. zooepidemicus was not mutated as seen in isolates of Streptococcus pyogenes with increased capsule production at 25°C.
  
  
  CONCLUSIONS
  Sensitivity of capsule expression to temperature above 35°C but not HylC by the general population of equine S. zooepidemicus indicates that capsule is not required for extended colonisation nor for opportunist invasion. Instead, capsule production at lower than body temperature may reflect adaptation to life on skin and mucosal surfaces, where hyaluronic acid contributes to adhesion and resistance to desiccation. Pathogenicity of S. zooepidemicus following opportunist invasion is possibly dependent on factors other than capsule that may be co-regulated with HylC. The Summary is available in Chinese - see Supporting information.
authors.affiliation: Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington, USA.
authors.forename: S
authors.initials: S
authors.lastname: Velineni
authors.forename: J F
authors.initials: JF
authors.lastname: Timoney
mesh_terms: D016667:Bacterial Capsules / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D006820:Hyaluronic Acid / Q000235:genetics / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000235:genetics / Q000378:metabolism; D018502:Streptococcus equi / Q000235:genetics / Q000378:metabolism*; D013696:Temperature; D015854:Up-Regulation
chemical_list: D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase
keywords: Streptococcus zooepidemicus; capsular hyaluronic acid; hasA; horse; hylC