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Table 1.

Microbial colonization of untreated ossicles.

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Table 2.

Microbial colonization of ossicle specimens with and without exposure to HHP treatment (350 MPa, 10 minutes).

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Table 3.

Microbial viability of Gram-positive bacteria, Gram-negative bacteria and yeasts after HHP treatment (350 MPa, 10 minutes).

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Table 4.

Microbial colonization of ossicles after HHP treatment (350 MPa, 10 minutes) with and without the addition of antibiotics to the media.

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Figure 1.

In vitro biofilms of clinical isolates before HHP treatment.

Biofilms of S. epidermidis (A) and N. subflava (B) after 4 days of in vitro growth (SEM pictures, magnification ×5000).

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Figure 1 Expand

Figure 2.

Viability of planktonic Staphylococcus epidermidis cells after HHP treatment using increasing levels of pressure.

Pressure levels of 100 MPa, 250 MPa, 400 MPa and 540 MPa were used. For HHP treatment, planktonic S. epidermidis cells were suspended in 0.9% NaCl (circles and small diamonds) or H2O (triangles and large squares). CFU: colony forming units as determined by viability counts. The figure shows the results of two representative and independent assays.

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Figure 2 Expand

Figure 3.

Viability of planktonic Neisseria subflava cells after HHP treatment using increasing levels of pressure.

Pressure levels of 100 MPa, 250 MPa, 400 MPa and 540 MPa were used. For HHP treatment, planktonic N. subflava cells were suspended in 0.9% NaCl (circles and small diamonds) or H2O (triangles and large squares). CFU: colony forming units as determined by viability counts. The figure shows the results of two representative and independent assays.

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Figure 3 Expand

Figure 4.

Effects of high hydrostatic pressure treatment on planktonic bacteria in isotonic medium.

REM [A–D] and TEM [E, F] pictures (magnification ×10 000). S. epidermidis (A, B, E, F) and N. subflava (C, D) cells were exposed to 100 MPa for 10 minutes (A, C, E) and to 540 MPa for 10 minutes (B, D, F). Whereas cell morphology appears to be unaffected after exposure to 100 MPa, both isolates show morphological changes after exposure to 540 MPa.

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Figure 4 Expand

Figure 5.

Viability of Staphylococcus epidermidis cells within biofilms after HHP treatment using increasing levels of pressure.

Pressure levels of 100 MPa, 250 MPa, 400 MPa and 540 MPa were used. S. epidermidis biofilms were grown for 3 days. For HPP treatment, the cells were suspended in 0.9% NaCl (circles and small diamonds) or H2O (triangles and large squares). CFU: colony forming units as determined by viability counts. The figure shows the results of two representative and independent assays.

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Figure 5 Expand

Figure 6.

Viability of Neisseria subflava cells within biofilms after HHP treatment using increasing levels of pressure.

Pressure levels of 100 MPa, 250 MPa, 400 MPa and 540 MPa were used. N. subflava biofilms were grown for 3 days. For HPP treatment, the cells were suspended in 0.9% NaCl (circles and small diamonds) or H2O (triangles and large squares). CFU: colony forming units as determined by viability counts. The figure shows the results of two representative and independent assays.

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Figure 6 Expand

Figure 7.

Effects of high hydrostatic pressure treatment on an in vitro S. epidermidis biofilm after 3 days of growth.

SEM pictures. A, B: intact cell and extracellular matrix morphology before pressure treatment, magnification ×500 (A) and ×5000 (B). C, D: destruction zones with only few structurally intact cells after pressure treatment at 540 MPa for 10 minutes, magnification ×500 (C) and ×5000 (D).

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Figure 7 Expand

Figure 8.

Effects of high hydrostatic pressure treatment on an in vitro N. subflava biofilm after 3 days of growth.

SEM pictures. A, B: intact cell and extracellular matrix morphology before pressure treatment, magnification ×1000 (A) and ×5000 (B). C, D: destruction zones with only few structurally intact cells after pressure treatment at 540 MPa for 10 minutes, magnification ×1000 (C) and ×5000 (D).

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Figure 8 Expand