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Residual antibiotics in decontaminated human cardiovascular tissues intended for transplantation and risk of falsely negative microbiological analyses.

Buzzi M, Guarino A, Gatto C, Manara S, Dainese L, Polvani G, Tóthová JD - PLoS ONE (2014)

Bottom Line: The RESEP tube treatment markedly reduced or totally eliminated the antimicrobial activity of tested tissues and media.After thawing, none of the 16 cryopreserved samples resulted positive with direct inoculum method.When the same samples were tested after removal of antibiotic residues, 8 out of 16 (50%) were contaminated.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Tissue Bank of Emilia-Romagna, Azienda Ospedaliero-Universitaria Policlinico S. Orsola-Malpighi, Bologna, Italy.

ABSTRACT
We investigated the presence of antibiotics in cryopreserved cardiovascular tissues and cryopreservation media, after tissue decontamination with antibiotic cocktails, and the impact of antibiotic residues on standard tissue bank microbiological analyses. Sixteen cardiovascular tissues were decontaminated with bank-prepared cocktails and cryopreserved by two different tissue banks according to their standard operating procedures. Before and after decontamination, samples underwent microbiological analysis by standard tissue bank methods. Cryopreserved samples were tested again with and without the removal of antibiotic residues using a RESEP tube, after thawing. Presence of antibiotics in tissue homogenates and processing liquids was determined by a modified agar diffusion test. All cryopreserved tissue homogenates and cryopreservation media induced important inhibition zones on both Staphylococcus aureus- and Pseudomonas aeruginosa-seeded plates, immediately after thawing and at the end of the sterility test. The RESEP tube treatment markedly reduced or totally eliminated the antimicrobial activity of tested tissues and media. Based on standard tissue bank analysis, 50% of tissues were found positive for bacteria and/or fungi, before decontamination and 2 out of 16 tested samples (13%) still contained microorganisms after decontamination. After thawing, none of the 16 cryopreserved samples resulted positive with direct inoculum method. When the same samples were tested after removal of antibiotic residues, 8 out of 16 (50%) were contaminated. Antibiotic residues present in tissue allografts and processing liquids after decontamination may mask microbial contamination during microbiological analysis performed with standard tissue bank methods, thus resulting in false negatives.

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Related in: MedlinePlus

Presence of antibiotic residues in decontaminated and cryopreserved tissue homogenates and cryopreservation media.Agar diffusion assays performed using Staphylococcus aureus-seeded plates (A) and Pseudomonas aeruginosa-seeded plates (B), immediately after thawing (initial) and after 14 days of sterility testing (final) on tissues and media undergoing decontamination procedures with Cocktail 1 and Cocktail 2, prepared by tissue banks, and BASE.128 as a control. The plot legend on Figure 1A also applies to Figure 1B.
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pone-0112679-g001: Presence of antibiotic residues in decontaminated and cryopreserved tissue homogenates and cryopreservation media.Agar diffusion assays performed using Staphylococcus aureus-seeded plates (A) and Pseudomonas aeruginosa-seeded plates (B), immediately after thawing (initial) and after 14 days of sterility testing (final) on tissues and media undergoing decontamination procedures with Cocktail 1 and Cocktail 2, prepared by tissue banks, and BASE.128 as a control. The plot legend on Figure 1A also applies to Figure 1B.

Mentions: Antibiotic elimination from tissue samples with the RESEP tube reduced inhibition zones by 81%–98% and 94%–100% on initial and final day of the 14-day sterility testing period, respectively (Figure 1A). Similarly, all cryopreservation media inhibited the growth of SA on agar plates. Cocktail 1, cocktail 2, and BASE.128 residues induced 11.2-, 6.9-, and 7.1-mm inhibition zones on SA plates, respectively. All cryopreservation media inoculated in growth broths induced detectable SA inhibition activity that was totally abolished by the RESEP tube treatment (Figure 1A).


Residual antibiotics in decontaminated human cardiovascular tissues intended for transplantation and risk of falsely negative microbiological analyses.

Buzzi M, Guarino A, Gatto C, Manara S, Dainese L, Polvani G, Tóthová JD - PLoS ONE (2014)

Presence of antibiotic residues in decontaminated and cryopreserved tissue homogenates and cryopreservation media.Agar diffusion assays performed using Staphylococcus aureus-seeded plates (A) and Pseudomonas aeruginosa-seeded plates (B), immediately after thawing (initial) and after 14 days of sterility testing (final) on tissues and media undergoing decontamination procedures with Cocktail 1 and Cocktail 2, prepared by tissue banks, and BASE.128 as a control. The plot legend on Figure 1A also applies to Figure 1B.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232473&req=5

pone-0112679-g001: Presence of antibiotic residues in decontaminated and cryopreserved tissue homogenates and cryopreservation media.Agar diffusion assays performed using Staphylococcus aureus-seeded plates (A) and Pseudomonas aeruginosa-seeded plates (B), immediately after thawing (initial) and after 14 days of sterility testing (final) on tissues and media undergoing decontamination procedures with Cocktail 1 and Cocktail 2, prepared by tissue banks, and BASE.128 as a control. The plot legend on Figure 1A also applies to Figure 1B.
Mentions: Antibiotic elimination from tissue samples with the RESEP tube reduced inhibition zones by 81%–98% and 94%–100% on initial and final day of the 14-day sterility testing period, respectively (Figure 1A). Similarly, all cryopreservation media inhibited the growth of SA on agar plates. Cocktail 1, cocktail 2, and BASE.128 residues induced 11.2-, 6.9-, and 7.1-mm inhibition zones on SA plates, respectively. All cryopreservation media inoculated in growth broths induced detectable SA inhibition activity that was totally abolished by the RESEP tube treatment (Figure 1A).

Bottom Line: The RESEP tube treatment markedly reduced or totally eliminated the antimicrobial activity of tested tissues and media.After thawing, none of the 16 cryopreserved samples resulted positive with direct inoculum method.When the same samples were tested after removal of antibiotic residues, 8 out of 16 (50%) were contaminated.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Tissue Bank of Emilia-Romagna, Azienda Ospedaliero-Universitaria Policlinico S. Orsola-Malpighi, Bologna, Italy.

ABSTRACT
We investigated the presence of antibiotics in cryopreserved cardiovascular tissues and cryopreservation media, after tissue decontamination with antibiotic cocktails, and the impact of antibiotic residues on standard tissue bank microbiological analyses. Sixteen cardiovascular tissues were decontaminated with bank-prepared cocktails and cryopreserved by two different tissue banks according to their standard operating procedures. Before and after decontamination, samples underwent microbiological analysis by standard tissue bank methods. Cryopreserved samples were tested again with and without the removal of antibiotic residues using a RESEP tube, after thawing. Presence of antibiotics in tissue homogenates and processing liquids was determined by a modified agar diffusion test. All cryopreserved tissue homogenates and cryopreservation media induced important inhibition zones on both Staphylococcus aureus- and Pseudomonas aeruginosa-seeded plates, immediately after thawing and at the end of the sterility test. The RESEP tube treatment markedly reduced or totally eliminated the antimicrobial activity of tested tissues and media. Based on standard tissue bank analysis, 50% of tissues were found positive for bacteria and/or fungi, before decontamination and 2 out of 16 tested samples (13%) still contained microorganisms after decontamination. After thawing, none of the 16 cryopreserved samples resulted positive with direct inoculum method. When the same samples were tested after removal of antibiotic residues, 8 out of 16 (50%) were contaminated. Antibiotic residues present in tissue allografts and processing liquids after decontamination may mask microbial contamination during microbiological analysis performed with standard tissue bank methods, thus resulting in false negatives.

Show MeSH
Related in: MedlinePlus