Phage Milagro: a platform for engineering a broad host range virulent phage for Burkholderia.

IMPORTANCE: Burkholderia infections are a significant concern in people with CF and other immunocompromising disorders, and are difficult to treat with conventional antibiotics due to their inherent drug resistance. Bacteriophages, or bacterial viruses, are now seen as a potential alternative therapy for these infections, but most of the naturally occurring phages are temperate and have narrow host ranges, which limit their utility as therapeutics. Here we describe the temperate Burkholderia phage Milagro and our efforts to engineer this phage into a potential therapeutic by expanding the phage host range and selecting for phage mutants that are strictly virulent. This approach may be used to generate new therapeutic agents for treating intractable infections in CF patients.

Complete Genome Sequence of Burkholderia cenocepacia Phage Paku.

Burkholderia cenocepacia is able to cause infections in cystic fibrosis patients. B. cenocepacia phage Paku has a 42,727-bp genome sharing a phiKMV-like genome arrangement. T7-like tail components were identified in parallel with a tyrosine integrase, suggesting that Paku might exhibit a temperate lifestyle, an atypical feature for an Autographiviridae phage.

Complete Genome Sequence of Burkholderia gladioli Myophage Mana.

Burkholderia gladioli is known to cause respiratory tract infections in cystic fibrosis patients. Here, we describe the annotation of the 38,038-bp genome sequence of Mana, a P2-like phage of B. gladioli Understanding the genomic characteristics of phages infecting pathogens like B. gladioli can lead to advancements in phage therapy.

Complete Genome Sequence of Burkholderia cenocepacia Phage Mica.

Burkholderia cenocepacia is a multidrug-resistant Gram-negative pathogen known to colonize patients with chronic granulomatous disease and cystic fibrosis. Here, we describe Burkholderia phage Mica, which is predicted to be a lysogenic myophage based on the similarity of its structural proteins to Enterobacteria phage P2 and Burkholderia phage KL3.

Complete Genome Sequence of Burkholderia gladioli Phage Maja.

Burkholderia gladioli is a Gram-negative bacterium associated with cystic fibrosis infections. Here, we describe the genome sequence of B. gladioli phage Maja. Maja is most related to another Burkholderia phage, BcepF1, and may be a temperate phage, despite the absence of repressor or integrase homologs in its genome sequence.

Complete Genome Sequence of Achromobacter xylosoxidans Myophage Mano.

Achromobacter spp. are ubiquitous Gram-negative bacteria, some of which can cause respiratory tract infections in patients with autoimmune disorders and cystic fibrosis. Bacteriophages have therapeutic and biotechnological potential to combat Achromobacter sp. infections. This announcement details the 42.5-kb genome sequence of the temperate Achromobacter xylosoxidans myophage Mano.

A Broad-Host-Range Tailocin from Burkholderia cenocepacia.

The Burkholderia cepacia complex (Bcc) consists of 20 closely related Gram-negative bacterial species that are significant pathogens for persons with cystic fibrosis (CF). Some Bcc strains are highly transmissible and resistant to multiple antibiotics, making infection difficult to treat. A tailocin (phage tail-like bacteriocin), designated BceTMilo, with a broad host range against members of the Bcc, was identified in B. cenocepacia strain BC0425. Sixty-eight percent of Bcc representing 10 species and 90% of non-Bcc Burkholderia strains tested were sensitive to BceTMilo. BceTMilo also showed killing activity against Pseudomonas aeruginosa PAO1 and derivatives. Liquid chromatography-mass spectrometry analysis of the major BceTMilo proteins was used to identify a 23-kb tailocin locus in a draft BC0425 genome. The BceTMilo locus was syntenic and highly similar to a 24.6-kb region on chromosome 1 of B. cenocepacia J2315 (BCAL0081 to BCAL0107). A close relationship and synteny were observed between BceTMilo and Burkholderia phage KL3 and, by extension, with paradigm temperate myophage P2. Deletion mutants in the gene cluster encoding enzymes for biosynthesis of lipopolysaccharide (LPS) in the indicator strain B. cenocepacia K56-2 conferred resistance to BceTMilo. Analysis of the defined mutants in LPS biosynthetic genes indicated that an α-d-glucose residue in the core oligosaccharide is the receptor for BceTMilo.IMPORTANCE BceTMilo, presented in this study, is a broad-host-range tailocin active against Burkholderia spp. As such, BceTMilo and related or modified tailocins have potential as bactericidal therapeutic agents against plant- and human-pathogenic Burkholderia.