Journal Article

Survival of Rickettsia conorii in artificially contaminated whole and leukoreduced canine blood units during the storage period

Wurzburger, Wendy, Toniolo, Federica, Mion, Monica, Lucchese, Laura, Da Rold, Graziana, Vascellari, Marta, Ravagnan, Silvia, Carminato, Antonio, Capelli, Gioia, Natale, Alda, Fournier, Pierre-Edouard

2020

Overview

Background The ability of tick-borne agents to survive in stored blood bags is a key factor for their transmissibility by blood transfusion. The aim of this study was to evaluate the survival and potential infectivity of Rickettsia conorii (RC) in artificially contaminated canine whole blood (WB) and in leukoreduced whole blood (LR-WB) during the storage period. Methods RC was cultured on L929 cells. We used a one-week 25-cm 2 flask with 70–80% of L929 infected cells to prepare the bacterial inoculum by pelleting cells and suspending the pellet in the donors’ serum. We infected five 100 ml WB units with RC within 2 h from the collection and maintained it at room temperature for 4 h prior to refrigeration. We filtered 50 ml of each WB bag to obtain leukoreduced WB (LR-WB) at day 1 post-infection (dpi). We checked WB and LR-WB bags at 1, 4, 7, 14, 21, 28, 35 dpi for RC presence and viability through real-time PCR (rPCR) for DNA and mRNA, respectively, and by isolation. Identification of isolates was confirmed by indirect immunofluorescence and rPCRs. Results RC survived for the entire storage period in both whole and leukoreduced blood. All bags contained viable bacteria until 7 dpi; RC viability generally decreased over time, particularly in LR-WB bags where the isolation time was longer than in WB. Viable bacteria were still isolated at 35 dpi in 3 WB and 3 LR-WB. Conclusions Leukoreduction reduced but did not eliminate RC in infected units. The survival and infectivity of RC in canine blood during the storage period may represent a threat for recipients.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: 15th Symposium on Canine Vector-Borne Diseases (CVBD) / ambient temperature / Animals / Arachnids / Bacteria / Bacteriology / bags / Biomedical and Life Sciences / Biomedicine / Blood / Blood & organ donations / Blood - microbiology / Blood Culture - veterinary / Blood Preservation - veterinary / blood serum / Blood Specimen Collection - veterinary / Blood transfusion / Blood Transfusion - veterinary / Boutonneuse Fever - microbiology / Boutonneuse Fever - prevention & control / Boutonneuse Fever - transmission / Cardiology and cardiovascular system / Cells / Deoxyribonucleic acid / DNA / DNA, Bacterial - genetics / Dog / Dogs / Emerging diseases / Entomology / Erythrocytes - microbiology / fluorescent antibody technique / Human health and pathology / Immunofluorescence / Infections / Infectious Diseases / Infectivity / Inoculum / Laboratories / Leukoreduction / Life Sciences / Medicine / messenger RNA / Microbiology and Parasitology / mRNA / Nucleotide sequence / Parasitology / pathogenicity / Pathogens / PCR / Pelleting / quantitative polymerase chain reaction / Refrigeration / Rickettsia conorii / Rickettsia conorii - genetics / Rickettsia conorii - physiology / RNA / Room temperature / Serum / Storage / storage time / Survival / Tick-borne diseases / Transfusion / Tropical Medicine / Veterinary Medicine/Veterinary Science / viability / Virology