title: Sub-MICs of Carum copticum and Thymus vulgaris influence virulence factors and biofilm formation in Candida spp.
abstract: BACKGROUND
  Emergence of drug-resistant strains of Candida and inefficiency of conventional antifungal therapy has necessitated the search for alternative and new antifungal agents. Inhibition of virulence and biofilm are the potential drug targets. In this study, the oils of Carum copticum, Thymus vulgaris and their major active compound thymol as revealed by Gas chromatography and gas chromatography-mass spectrometry (GC-GC/MS) analysis were tested for their inhibitory activity against growth to determine sub-MIC values against 27 drug-resistant strains of Candida spp.
  
  
  METHODS
  Brothmacrodilution method was used for determination of MIC of test oils against Candida strains. The spectrophotometric methods were used for detection and inhibition assays for virulence factors in Candida spp. Light and electron microscopy was performed to observe morphological effects of oils on biofilms. GC-GC/MS were used to evaluate the major active compounds of test oils.
  
  
  RESULTS
  Virulence factors like proteinase and haemolysin were detected in 18 strains, both in solid and liquid media. A 70% of the test strains exhibited hydrophobicity and formed moderate to strong biofilms (OD280 0.5- > 1.0). Test oils exhibited MICs in the range of 45-360 μg.mL(-1) against the majority of test strains. All the oils at 0.25× and 0.5× MICs induced >70% reduction in the cell surface hydrophobicity, proteinase and haemolysin production. At 0.5× MIC, thymol and T. vulgaris were most inhibitory against biofilm formation. At sub-MICs electron microscopic studies revealed the deformity of complex structures of biofilms formed and cell membranes appeared to be the target site of these agents.
  
  
  CONCLUSIONS
  Therefore, our findings have highlighted the concentration dependent activity of oils of C. copticum and T. vulgaris against virulence factors and biofilms in proteinase and haemolysin producing drug-resistant strains of Candida spp. The above activities of test oils are supposed to be mainly contributed due to their major active compound thymol. Further mechanism involving anti-proteinase, anti-haemolysin and anti-biofilm activities of these oils and compounds are to be explored for possible exploitation in combating Candida infections.
authors.affiliation: Phytomedicine Programme, Department of Paraclinical Sciences, University of Pretoria, Pretoria 0110, South Africa. khanmsa@hotmail.com.
authors.forename: Mohd S A
authors.initials: MS
authors.lastname: Khan
authors.forename: Iqbal
authors.initials: I
authors.lastname: Ahmad
authors.forename: Swaranjit S
authors.initials: SS
authors.lastname: Cameotra
authors.forename: Francien
authors.initials: F
authors.lastname: Botha
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D002175:Candida / Q000187:drug effects* / Q000378:metabolism; D029974:Carum / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry; D006461:Hemolysis / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D010936:Plant Extracts / Q000494:pharmacology*; D012756:Sheep; D046930:Thymus Plant / Q000737:chemistry*; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D000935:Antifungal Agents; D010936:Plant Extracts; D037521:Virulence Factors