Prevalence of a cefazolin inoculum effect associated with blaZ gene types, and clinical outcomes among methicillin-susceptible Staphylococcus aureus blood isolates of patients with infective endocarditis.

OBJECTIVES: A proportion of blaZ gene-positive methicillin-susceptible Staphylococcus aureus (MSSA) strains exhibits the cefazolin inoculum effect (CInE). Its clinical impact remains uncertain but could compromise the use of cefazolin in high-burden infections. To date, no study has been conducted in France or in Europe. We aimed to assess the prevalence of CInE and its association with blaZ beta-lactamase and S. aureus protein A (spa) types, and to assess the clinical outcomes in cefazolin-treated patients for infective endocarditis whose strain exhibited a CInE. METHODS: This was a French single-center retrospective study of 51 MSSA strains from patients of the Nantes endocarditis prospective cohort, conducted between 2013 and 2018. RESULTS: Cefazolin MIC50 at high inoculum was 2 mg/L (IQR 1-2). CInE was found in 17.6 % of tested strains. Among blaZ-positive strains (n = 44), type A beta-lactamase was predominant (n = 25, 57 %). Thirty-seven S. aureus protein A (spa) types were found. No statistical association was shown between blaZ or spa types and CInE. CInE was neither associated with a higher rate of persistent bacteremia (25 % vs 56.3 %, p = 0.58) nor with clinical failure in patients treated with cefazolin, in comparison to patients with no CInE strain (25 % vs 56.3 %, p = 0.58). CONCLUSION: The cefazolin inoculum effect was found in a substantial number of Staphylococcus aureus strains; however, minimum inhibitory concentrations remained globally low. CInE was not associated with a higher proportion of clinical failure on treatment.

New in vitro and in vivo models to evaluate antibiotic efficacy in Staphylococcus aureus prosthetic vascular graft infection.

OBJECTIVE: Prosthetic vascular graft infection (PVGI) is an emerging disease, mostly caused by staphylococci, with limited data regarding efficacy of current antistaphylococcal agents. We aimed to assess the efficacy of different antibiotic regimens. METHODS: Six different strains of MSSA and MRSA were used. We compared results of minimal biofilm inhibitory and eradicating concentrations (MBICs and MBECs) obtained with a Calgary Biofilm Pin Lid Device (CBPD) with those yielded by an original Dacron(®)-related minimal inhibitory and eradicating concentration measure model. We then used a murine model of Staphylococcus aureus vascular prosthetic material infection to evaluate efficacy of different antibiotic regimens: vancomycin and daptomycin combined or not with rifampicin for MRSA and the same groups with cloxacillin and cloxacillin combined with rifampicin for MSSA. RESULTS: We demonstrated that classical measures of MBICs and MBECs obtained with a CPBD could overestimate the decrease in antibiotic susceptibility in material-related infections and that the nature of the support used might influence the measure of biofilm susceptibility, since results yielded by our Dacron(®)-related minimal eradicating assay were lower than those found with a plastic device. In our in vivo model, we showed that daptomycin was significantly more bactericidal than comparators for some strains of MRSA or MSSA but not for all. For the majority of strains, it was as efficient as comparators. The addition of rifampicin to daptomycin did not enhance daptomycin efficacy. CONCLUSIONS: Despite the heterogeneity of results according to bacterial strains, these innovative models represent an option to better evaluate the in vitro efficacy of antibiotics on Dacron(®)-related biofilm S. aureus infections, and to screen different antibiotic regimens in a mouse model of PVGIs.

A delivery system of linezolid to enhance the MRSA osteomyelitis prognosis: in vivo experimental assessment.

Staphylococcus aureus, a major responsible microorganism of osteomyelitis, represents a challenge to treat because of the poor penetration of antibiotics in bone and increasing minimum inhibitory concentrations (MICs) to glycopeptides. The calcium-deficient apatites (CDA), closer to the biological components found in bone and other calcified tissues, have osteoconductive properties. So, to process severe osseous infections, CDA can be used to deliver in the infectious site antibiotics like linezolid. The acute experimental osteomyelitis due to methicillin-resistant Staphylococcus aureus (MRSA) was induced in rabbit's femurs and surgery mimicking human procedures was performed at day three after inoculation. Animals were randomly assigned to treatment groups: L((IV)) [4-day linezolid IV infusion, human-equivalent dose of 10 mg/kg/12 h], L((CDA50%)) (100 mg CDA with linezolid 500 µg/mg) and L((CDA50%)) + L((IV)). Surviving bacteria were counted in bone marrow (BM) and bone (Bo) at day 3 (before treatment), day 7 (4-day treatment) or day 17 (14-day treatment). L(iv) was effective after a 4-day treatment with a log(10)CFU/g decrease of -2.63 ± 1.92 and -2.17 ± 1.58 in bone marrow and bone, respectively. CDA loaded with linezolid enhance the efficacy of the IV linezolid regimen by more than one log(10)CFU/g.

A new experimental model of acute osteomyelitis due to methicillin-resistant Staphylococcus aureus in rabbit.

AIMS: To assess the impact of antibiotic therapy on severe osseous infections, animal models of chronic bacterial infections have been developed; however, these models suffer from many experimental limitations. The aim of this work was to develop a new model system in which high levels of bacteria are obtained within femoral bone marrow and bone tissue, and such infections are maintained for at least 14 days. METHODS AND RESULTS: Experimental osteomyelitis was induced in 25 New Zealand white rabbits. A 10(9) CFU ml(-1) suspension of methicillin-resistant Staphylococcus aureus was injected into the knee after bone trepanation. On day 3, surgical debridement was performed to mimic a surgical procedure. Animals were euthanized 1, 2, 3, 9 and 14 days post-inoculation to determine the bacterial counts in marrow and bone, and to evaluate the stability of the infection. Inoculated lesions also were assessed for changes in histological parameters on days 3 and 7 post-inoculation. At days 1, 2, 3, 9 and 14 post-inoculation, we observed 6·50 ± 0·64, 7·30 ± 0·49, 7·82 ± 0·19, 8·00 ± 1·48 and 8·99 ± 0·20 log10 CFU g(-1) in bone marrow and 8·40 ± 0·68, 7·65 ± 0·27, 7·58 ± 0·30, 8·88 ± 0·52 and 8·28 ± 0·39 log10 CFU g(-1) in bone tissue, respectively. No statistical differences in bacterial count were found between bone marrow and bone tissue at any time point. CONCLUSION: This new model of acute osteomyelitis was validated by histological and microbiological changes in the absence of sclerosing agents, and these changes remained stable for 14 days. SIGNIFICANCE AND IMPACT OF THE STUDY: These results describe a new experimental model of acute osteomyelitis and demonstrate its usefulness in assessing the activity of antibacterial agents in vivo soon after bone infection.

Efficacy of daptomycin combined with rifampicin for the treatment of experimental meticillin-resistant Staphylococcus aureus (MRSA) acute osteomyelitis.

Daptomycin exhibits rapid bactericidal activity against Gram-positive organisms, including meticillin-resistant Staphylococcus aureus (MRSA). Daptomycin in combination with rifampicin needs to be assessed in bone infection. An MRSA acute osteomyelitis model was used. Daptomycin and vancomycin were compared, alone or in combination with rifampicin, over 4 days. Surviving bacteria were counted in bone, bone marrow and joint fluid. Vancomycin and daptomycin as single therapies were ineffective, but both combinations were significantly more effective than the corresponding monotherapy. Combination of daptomycin and rifampicin could prevent S. aureus from developing resistance. This combination could be a useful alternative to treat MRSA osteomyelitis at an early stage.

In vivo assessment of the antimicrobial activity of a calcium-deficient apatite vancomycin drug delivery system in a methicillin-resistant Staphylococcus aureus rabbit osteomyelitis experimental model.

The antimicrobial activities of calcium-deficient apatite loaded with different concentrations (25, 100, and 500 microg/mg) of vancomycin as a filling biomaterial were evaluated in a methicillin-resistant Staphylococcus aureus (MRSA) rabbit acute osteomyelitis model. Bacterial counts in bone, bone marrow, and joint fluid samples treated with forms of the apatite were compared to those in tissue samples receiving a constant intravenous vancomycin infusion after 4 days. This study demonstrates that using a calcium-deficient apatite loaded with vancomycin dramatically decreases the bacterial counts in bone and marrow.

[In vivo activity of amoxicillin in a non-toxigenic Corynebacterium diphtheriae rabbit endocarditis experimental model]. / Activité de l'amoxicilline in vivo dans l'endocardite expérimentale du lapin à Corynebacterium diphtheriae non toxinogène.

The mortality of non-toxigenic Corynebacterium diphtheriae endocarditis is high (27%). One explanation could be tolerance to amoxicillin. The aim of this work was to evaluate in vivo the tolerance phenomenon, in a rabbit endocarditis experimental model. Two strains of non-toxigenic Corynebacterium diphtheriae were compared: a tolerant one and a non-tolerant one. EACH ANIMAL WAS RANDOMLY ASSIGNED TO ONE OF THE FOLLOWING GROUPS: no treatment, continuous infusion of amoxicillin simulating 200 mg/kg/24 h in humans, or fractionated infusion of amoxicillin simulating 66 mg/kg/8 h in humans. Surviving bacteria were counted in vegetations after one or three days of treatment. The 24 h fractionated amoxicillin infusion was more efficacious on the non-tolerant strain than on the tolerant strain. On the tolerant strain, 24 h amoxicillin was more efficacious as a continuous infusion than as a fractionated one.