title: Recovery of Leptospires in Short- and Medium-Term Cryopreservation Using Different Glycerol and Dimethyl Sulfoxide Concentrations.
abstract: Cryopreservation is a recognized method for the maintenance of Leptospira collections. Although cryoprotectants are commonly used in order to prevent or reduce the adverse effects of freezing, there is no consensus regarding the protocols of cryopreservation. This study aimed to compare cryopreservation protocols for Leptospira using different glycerol and dimethyl sulfoxide (DMSO) concentrations. Leptospira interrogans serovar Icterohaemorrhagiae, L. interrogans serovar Bratislava, and L. borgpetersenii serovar Hardjo were used as the experimental strains. For each strain, three protocols were tested using 5% and 10% glycerol and 2.5% DMSO. For each protocol, 12 tubes containing 1.5 mL of serovar were frozen at -70°C on the same day. An aliquot of each serovar/protocol was thawed once a month throughout 1 year. The viability of leptospires was evaluated by the recovery of those at days 7, 14, and 21 after thawing. Although no significant difference was found among the leptospiral recovery rates for the 9 serovar/protocols tested, DMSO (2.5%) was shown to be slightly better than glycerol, and its use should be encouraged as a cryoprotectant for leptospires.
authors.affiliation: 1 Laboratory of Veterinary Bacteriology, Department of Microbiology and Parasitology, Universidade Federal Fluminense , Rio de Janeiro, Brazil .
authors.forename: Lorena
authors.initials: L
authors.lastname: Narduche
authors.affiliation: 1 Laboratory of Veterinary Bacteriology, Department of Microbiology and Parasitology, Universidade Federal Fluminense , Rio de Janeiro, Brazil .
authors.forename: Camila
authors.initials: C
authors.lastname: Hamond
authors.affiliation: 1 Laboratory of Veterinary Bacteriology, Department of Microbiology and Parasitology, Universidade Federal Fluminense , Rio de Janeiro, Brazil .
authors.forename: Gabriel M S
authors.initials: GM
authors.lastname: Martins
authors.affiliation: 2 Bio-Manguinhos, Oswaldo Cruz Foundation, Brazilian Ministry of Health , Rio de Janeiro, Brazil .
authors.forename: Marco A
authors.initials: MA
authors.lastname: Medeiros
authors.affiliation: 1 Laboratory of Veterinary Bacteriology, Department of Microbiology and Parasitology, Universidade Federal Fluminense , Rio de Janeiro, Brazil .
authors.forename: Walter
authors.initials: W
authors.lastname: Lilenbaum
mesh_terms: D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D007919:Leptospira / Q000187:drug effects; D007921:Leptospira interrogans / Q000187:drug effects
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide