title: An insight into the antibiofilm properties of Costa Rican stingless bee honeys.
abstract: OBJECTIVE
  There is an increasing search for antibiofilm agents that either have specific activity against biofilms or may act in synergy with antimicrobials. Our objective is to examine the the antibiofilm properties of stingless bee honeys.
  
  
  METHODS
  Meliponini honeys from Costa Rica were examined along with Medihoney as a reference. All honeys were submitted to a screening composed of minimum inhibitory concentration, inhibition of biofilm formation and biofilm destruction microplate-based assays against a Staphylococcus aureus biofilm forming strain. Dialysis led to the isolation of an antibiofilm fraction in Tetragonisca angustula honeys. The honey antibiofilm fraction was evaluated for protease activity and for any synergistic effect with antibiotics on a Staphylococcus aureus biofilm. The active fraction was then separated through activity guided isolation techniques involving SDS-PAGEs, anion exchange and size exclusion fast protein liquid chromatographies. The fractions obtained and the isolated antibiofilm constituents were tested for amylase and DNase activity.
  
  
  RESULTS
  A total of 57 Meliponini honeys from Costa Rica were studied in this research. The honeys studied belonged to the Tetragonisca angustula (n=36) and Melipona beecheii (n=21) species. Costa Rican Tetragonisca angustula honeys can inhibit the planktonic growth, biofilm formation, and are capable of destroying a Staphylococcus aureus biofilm. The antibiofilm effect was observed in the protein fraction of Tetragonisca angustula honeys. The biofilm destruction proteins allowed ampicillin and vancomycin to recover their antimicrobial activity over a Staphylococcus aureus biofilm. The antibiofilm proteins are of bee origin, and their activity was not due to serine, cysteine or metalloproteases. There were 2 proteins causing the antibiofilm action; these were named the Tetragonisca angustula biofilm destruction factors (TABDFs). TABDF-1 is a monomeric protein of approximately 50kDa that is responsible of the amylase activity of Tetragonisca angustula honeys. TABDF-2 is a protein monomer of approximately 75kDa.
  
  
  CONCLUSIONS
  Tetragonisca angustula honeys from Costa Rica are a promising candidate for research and development of novel wound dressings focused on the treatment of acute and chronic Staphylococcus aureus biofilm wound infections.
authors.affiliation: Microbiologist, Coordinator of the Microbiology and Medicinal Chemistry Program, Medicinal Chemistry and Chemical Biology Program, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University. David de Wiedgebouw, P. O. B. 80082, 3508 TB Utrecht, The Netherlands; Centro de Investigaciones Apícolas Tropicales, Universidad Nacional. P. O. Box 475-3000 Heredia, Costa Rica.
authors.forename: L G
authors.initials: LG
authors.lastname: Zamora
authors.affiliation: Associate Professor, Medicinal Chemistry and Chemical Biology Program, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University. David de Wiedgebouw, P. O. B. 80082, 3508 TB Utrecht, The Netherlands.
authors.forename: C J
authors.initials: CJ
authors.lastname: Beukelman
authors.affiliation: Associate Professor, Medicinal Chemistry and Chemical Biology Program, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University. David de Wiedgebouw, P. O. B. 80082, 3508 TB Utrecht, The Netherlands.
authors.forename: A J J
authors.initials: AJ
authors.lastname: van den Berg
authors.affiliation: Senior Technician, Medical Microbiology, University Medical Center Utrecht, P. O. B. 85500, 3508 GA Utrecht, The Netherlands.
authors.forename: P C
authors.initials: PC
authors.lastname: Aerts
authors.affiliation: Research and Education Assistant, Medicinal Chemistry and Chemical Biology Program, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University. David de Wiedgebouw, P. O. B. 80082, 3508 TB Utrecht, The Netherlands.
authors.forename: H C
authors.initials: HC
authors.lastname: Quarles van Ufford
authors.affiliation: Assistant Professor, Coordinator of the Bacteriology Group, Medical Microbiology, University Medical Center Utrecht, P. O. B. 85500, 3508 GA Utrecht, The Netherlands; Laboratory of Phytopatology, Wageningen University. P. O. Box 16, 6700 A A Wageningen, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Nijland
authors.affiliation: Coordinator of the Laboratory of Food Microbiology, Facultad de Microbiología, Universidad de Costa Rica. P. O. Box 2060-1000 San José, Costa Rica.
authors.forename: M L
authors.initials: ML
authors.lastname: Arias
mesh_terms: D000667:Ampicillin / Q000494:pharmacology; D000681:Amylases; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001516:Bees; D018441:Biofilms / Q000187:drug effects*; D003364:Costa Rica; D003851:Deoxyribonucleases; D006722:Honey; D008826:Microbial Sensitivity Tests; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000254:growth & development; D014640:Vancomycin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D014640:Vancomycin; D000667:Ampicillin; D003851:Deoxyribonucleases; D000681:Amylases
keywords: Melipona beecheii; Staphylococcus aureus; Tetragonisca angustula; antibiofilm; stingless bee honey