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Orthopaedic Proceedings
Vol. 101-B, Issue SUPP_14 | Pages 88 - 88
1 Dec 2019
EFFICACY OF SB-1 BACTERIOPHAGE IN TREATING AND PREVENTING METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS IN A GALLERIA MELLONELLA MODEL OF IMPLANT-ASSOCIATED INFECTION
Luca MD Materazzi A Klatt A Bottai D Tavanti A Trampuz A
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Aim

To investigate the ability of the bacteriophage Sb-1 to treat and prevent implant-associated infections due to methicillin-resistant Staphylococcus aureus (MRSA) in Galleria mellonella larvae implanted with a K-wire.

Method

The stability of Sb-1 in G. mellonella larvae was investigated by injecting a phage titer of 108 PFU and evaluating the presence of Sb-1 in hemolymph at different time points. For infection experiments, sterile stainless-steel K-wires (4 mm, 0.6 mm Ø) were implanted into larvae. Two days after implant, larvae were infected with MRSA ATCC 43300 (1×105 CFU) and incubated at 37°C for further 2 days. Implanted-infected larvae were thus treated for 2 days (3×/day) with 10µL of: i) PBS; ii) Sb-1 (107 PFU); iii) Daptomycin (4mg/kg), iv) PBS (24h)/Daptomycin(24h); v) Sb-1(24h)/Daptomycin(24h). To evaluate the prophylactic efficacy of Sb-1, an experiment based on phages or vancomycin (10mg/kg) administration, followed by MRSA infection of implanted larvae was performed. Both two days post-infection and post-treatment, K-wires were explanted, and the material was sonicated and plated for MRSA colony counting.

Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_17 | Pages 9 - 9
1 Dec 2018
AN ANTISTAPHYLOCOCCAL BACTERIOPHAGE LYSATE TO TREAT DRUG-RESISTANT STAPHYLOCOCCUS AUREUS INVOLVED IN PROSTHETIC JOINT INFECTIONS: AN ALTERNATIVE STRATEGY
Di Luca M Klatt A Trampuz A
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Aim

Virulent bacteriophages are known to be an effective therapy against various human bacterial infections. The aims of the study are to evaluate i) the killing activity of an antistaphylococcal phage lysate (ASPL), available in the Czech Republic for topical application, against Staphylococcal aureus (Sa) strains isolated in orthopedic infections; ii) the antimicrobial activity of ASPL against biofilm-embedded cells of a methicillin-resistant Sa (MRSA) standard strain.

Method

The susceptibility of 25 MRSA and 18 methicillin-sensitive Sa (MSSA) strains to the ASPL was evaluated by spot assay. In addition, susceptibility of four laboratory MRSA strains, including ATCC 43300, ATCC 33591, Mu3 (MRSA/hetero vancomycin intermediate resistant Sa) and Mu50 (MRSA/vancomycin-resistant Sa) was also tested. The activity of ASPL against planktonic and biofilm-embedded MRSA ATCC 43300 was evaluated in real-time by isothermal microcalorimetry. The minimum heat inhibitory concentrations (MHIC) was defined as the lowest antimicrobial concentration leading to the lack of heat flow production after 24h for both planktonic and biofilm-embedded cells. The viability of bacterial cells was assessed by plating and colony counting. The minimum bactericidal concentration (MBC) was defined as the lowest antimicrobial concentration leading the reduction of 3 log CFU compared to the untreated control.

Orthopaedic Proceedings
Vol. 99-B, Issue SUPP_22 | Pages 36 - 36
1 Dec 2017
ISOLATION OF NEW LYTIC BACTERIOPHAGES FOR TREATMENT OF PROSTHETIC JOINT INFECTION
Trampuz A Klatt A Luca MD
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Aim

Phage therapy has attracted attention as a promising alternative treatment option for biofilm infections.

To establish a successful phage therapy, a comprehensive stock of different phages covering a broad bacterial spectrum is crucial. We screened human and environmental sources for presence of lytic phages against selected bacteria.

Methods

Saliva collected from 10 volunteers and 500 ml of sewage water were screened for the presence of lytic phages active against 20 clinical strains of Staphylococcus aureus and 10 of Escherichia coli, both isolated from patients with prosthetic joint infection. Laboratory strains of methicillin-resistant S. aureus (MRSA)*1 and E. coli*2 were also tested. Screening was performed plaque-assay to detect phages for different strains. Isolated plaques were collected and phages were enriched to determine their activity against their bacterial host strains. The activity of bacteriophages against adherent E. coli and MRSA was evaluated by crystal violet, staining bacterial biofilms grown on glass beads.