Antibiofilm Activity of Actinobacillus pleuropneumoniae Serotype 5 Capsular Polysaccharide.docVIP

AntibiofilmActivityofActinobacilluspleuropneumoniae Serotype5CapsularPolysaccharide MichaelT.Karwacki1,DanielE.Kadouri1,MeriemBendaoud1,EraA.Izano1,VandanaSampathkumar1, ThomasJ.Inzana2,JeffreyB.Kaplan3* 1DepartmentofOralBiology,NewJerseyDentalSchool,Newark,NewJersey,UnitedStatesofAmerica,2DepartmentofBiomedicalSciencesandPathobiology,Virginia- Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia, United States of America, 3Department of Biology, American University, Washington,DistrictofColumbia,UnitedStatesofAmerica Abstract Cell-free extracts isolated from colony biofilms of Actinobacillus pleuropneumoniae serotype 5 were found to inhibit biofilm formation by Staphylococcus aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans, but not by A. pleuropneumoniae serotype 5 itself,in a96-well microtiter plateassay. Physical andchemical analysesindicated thatthe antibiofilm activity in the extract was due to high-molecular-weight polysaccharide. Extracts isolated from a mutant strain deficient in the production of serotype 5 capsular polysaccharide did not exhibit antibiofilm activity. A plasmid harboring the serotype 5 capsule genes restored the antibiofilm activity in the mutant extract. Purified serotype 5 capsular polysaccharide also exhibited antibiofilm activity against S. aureus. A. pleuropneumoniae wild-type extracts did not inhibit S. aureus growth, but did inhibit S. aureus intercellular adhesion and binding of S. aureus cells to stainless steel surfaces. Furthermore, polystyrene surfaces coated with A. pleuropneumoniae wild-type extracts, but not with capsule-mutant extracts, resisted S. aureus biofilm formation. Our findings suggest that the A. pleuropneumoniae serotype 5 capsule inhibits cell-to-cell and cell-to-surface interactions of other bacteria. A. pleuropneumoniae serotype 5 capsular polysaccharide is one of a growing number of bacterial polysaccharides that exhibit broad-spectrum, nonbiocidal antibio