title: Ultrastructural effects and antibiofilm activity of LFchimera against Burkholderia pseudomallei.
abstract: Lactoferrin chimera (LFchimera), a hybrid peptide containing the two antimicrobial stretches of the innate immunity factor bovine lactoferrin, viz. LFampin265-284 and LFcin17-30, has strikingly high antimicrobial activity against the category B pathogen Burkholderia pseudomallei. The action mechanisms of LFchimera against B. pseudomallei is not fully understood. The aim of this study was to further investigate the effect of treated B. pseudomallei with LFchimera using (immune) electron microscopy. The effects of LFchimera on biofilm formation and against preformed biofilm of B. pseudomallei were also determined. After exposure to LFchimera, transmission electron microscopy revealed swelling of the periplasmic space of B. pseudomallei and a highly inhomogeneous electron density in the intracellular DNA region. Localization of LFchimera in B. pseudomallei using immunoelectron microscopy showed gold particles in intracellular structures without accumulation on the membranes. LFchimera also possessed stronger bactericidal activity than ceftazidime against B. pseudomallei grown in biofilm. Moreover, limited exposure of B. pseudomallei to LFchimera at subcidal concentration could reduce biofilm formation. Altogether, the results indicate that LFchimera possesses antibacterial and antibiofilm activities and can modulate B. pseudomallei colonization. Therefore, the efficacy of LFchimera merits further development of this agent for the therapy of melioidosis.
authors.affiliation: Biofilm Research Group, Faculty of Dentistry, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands.|School of Medical Science, University of Phayao, Mueang Phayao, Phayao, 56000, Thailand.
authors.forename: Aekkalak
authors.initials: A
authors.lastname: Puknun
authors.affiliation: Biofilm Research Group, Faculty of Dentistry, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.|Melioidosis Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Sakawrat
authors.initials: S
authors.lastname: Kanthawong
authors.affiliation: Biofilm Research Group, Faculty of Dentistry, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands.|Melioidosis Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.|Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Chitchanok
authors.initials: C
authors.lastname: Anutrakunchai
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands.
authors.forename: Kamran
authors.initials: K
authors.lastname: Nazmi
authors.affiliation: Core Facility Cellular Imaging/LCAM-AMC, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Wikky
authors.initials: W
authors.lastname: Tigchelaar
authors.affiliation: Core Facility Cellular Imaging/LCAM-AMC, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Kees A
authors.initials: KA
authors.lastname: Hoeben
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands.
authors.forename: Enno C I
authors.initials: EC
authors.lastname: Veerman
authors.affiliation: Department of Oral Biochemistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands.
authors.forename: Jan G M
authors.initials: JG
authors.lastname: Bolscher
authors.affiliation: Biofilm Research Group, Faculty of Dentistry, Khon Kaen University, Khon Kaen, 40002, Thailand. suvi_taw@kku.ac.th.|Melioidosis Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand. suvi_taw@kku.ac.th.|Department of Oral Diagnosis, Faculty of Dentistry, Khon Kaen University, Khon Kaen, 40002, Thailand. suvi_taw@kku.ac.th.
authors.forename: Suwimol
authors.initials: S
authors.lastname: Taweechaisupapong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D016957:Burkholderia pseudomallei / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure*; D002417:Cattle; D002442:Ceftazidime / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects; D008554:Melioidosis / Q000628:therapy; D008826:Microbial Sensitivity Tests; D008854:Microscopy, Electron
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; C557489:LFchimera peptide; D002442:Ceftazidime
keywords: (Immune) electron microscopy; Antimicrobial peptide; Biofilm; Burkholderia pseudomallei; LFchimera; Melioidosis