Effect of platelet additive solution on bacterial dynamics and their influence on platelet quality in stored platelet concentrates.

BACKGROUND: Platelet additive solutions (PASs) are an alternative to plasma for the storage of platelet concentrates (PCs). However, little is known about the effect of PAS on the growth dynamics of contaminant bacteria. Conversely, there have been no studies on the influence of bacteria on platelet (PLT) quality indicators when suspended in PAS. STUDY DESIGN AND METHODS: Eight buffy coats were pooled, split, and processed into PCs suspended in either plasma or PAS (SSP+, MacoPharma). PCs were inoculated with 10 and 100 colony-forming units (CFUs)/bag of either Serratia liquefaciens or Staphylococcus epidermidis. Bacterial growth was measured over 5 days by colony counts and bacterial biofilm formation was assayed by scanning electron microscopy and crystal violet staining. Concurrently, PLT markers were measured by an assay panel and flow cytometry. RESULTS: S. liquefaciens exhibited an apparent slower doubling time in plasma-suspended PCs (plasma-PCs). Biofilm formation by S. liquefaciens and S. epidermidis was significantly greater in PCs stored in plasma than in PAS. Although S. liquefaciens altered several PLT quality markers by Days 3 to 4 postinoculation in both PAS- and plasma-PCs, S. epidermidis contamination did not produce measurable PLT changes. CONCLUSIONS: S. liquefaciens can be detected more quickly in PAS-suspended PCs (PAS-PCs) than in plasma-PCs by colony counting. Furthermore, reduced biofilm formation by S. liquefaciens and S. epidermidis during storage in PAS-PCs increases bacteria availability for sampling detection. Culture-based detection remains the earliest indicator of bacterial presence in PAS-PCs, while changes of PLT quality can herald S. liquefaciens contamination when in excess of 10(8) CFUs/mL.

Assessment of biofilm-forming ability of coagulase-negative staphylococci isolated from contaminated platelet preparations in Canada.

BACKGROUND: Coagulase-negative staphylococci (CoNS) are the most prevalent bacterial contaminants of platelet (PLT) preparations and have been implicated in adverse transfusion reactions worldwide. The most frequently identified contaminant is Staphylococcus epidermidis, which is noted for its ability to maintain chronic hospital-acquired infections by forming biofilms as a chief virulence mechanism. STUDY DESIGN AND METHODS: Strains of S. epidermidis isolated from contaminated PLT preparations in Canada were distinguished via gene-specific polymerase chain reaction (PCR) with divIVA as a marker. Biofilm-forming ability was assessed by the presence of the gene icaD, slime production on Congo red agar, and biofilm formation on polystyrene surfaces. Production of polysaccharide intercellular adhesin (PIA) was resolved by immunofluorescence. RESULTS: Eight of the 13 (62%) CoNS isolates under study were identified as S. epidermidis. Of these, four strains (50%) were classified as strong biofilm producers. Three of the four biofilm-positive strains (75%) produced slime, harbored the icaD gene, and had positive expression of PIA. CONCLUSIONS: Despite the presumable commensal origin of the CoNS isolates, a large proportion of S. epidermidis strains demonstrated a potential for enhanced virulence. Identification of contaminant staphylococci as biofilm producers is thus relevant and informative with regard to treatment approach in the circumstance of inadvertent infection of a PLT recipient.

Staphylococcus epidermidis forms biofilms under simulated platelet storage conditions.

BACKGROUND: Staphylococcus epidermidis grows slowly in platelet (PLT) preparations compared to other bacteria, presenting the possibility of missed detection by routine screening. S. epidermidis is a leading cause of nosocomial sepsis, with virulence residing in its ability to establish chronic infections through production of slime layers, or biofilms, on biomedical devices. This study aims to establish biofilm formation (BF) as a mode of growth by S. epidermidis in PLT preparations. STUDY DESIGN AND METHODS: Biofilm-positive (BFpos) and -negative (BFneg) S. epidermidis strains were grown in whole blood-derived PLTs (WBDPs) and in glucose-rich medium (TSBg). An assay for BF was adapted for cultures grown in WBDPs or filtered WBDPs in polystyrene culture plates. Bacterial attachment to polyvinylchloride PLT bags and PLTs was examined by scanning electron microscopy. RESULTS: Both strains display similar growth profiles in WBDPs and TSBg. Unexpectedly, evidence of BF was observed on PLT bags and on PLTs directly, not only by the BFpos strain but also by the BFneg strain. The BFpos strain displayed greater plastic adherence than the BFneg strain in WBDPs (p < 0.05). BF by the BFneg strain was approximately 10-fold greater in WBDPs compared to TSBg (p < 0.05), likely by use of PLTs as a scaffold. Furthermore, BF by S. epidermidis was significantly decreased when PLT concentration was reduced 1000-fold. CONCLUSIONS: S. epidermidis forms biofilms on PLT aggregates and on PLT bags under PLT storage conditions. Our results demonstrate that the PLT storage environment can promote a BF growth mechanism for contaminant bacteria.