Influência de crioprotetores e pré-adaptação na viabilidade e produção de transcritos por cepas de Campylobacter jejuni mantidas a -20°c

Abstract

Campylobacter is considered a fragile microorganism ans sensitive to environmental conditions, but demostrate strategies to survive in unfavorable environmental conditions. This study evaluated the viability and production of transcipts of the genes sodB, p19 ciaB and dnaJ in strains ATCC 33291, NCTC 11351 and IAL 2383 stored in UHT milk and neopeptona + 12% glycerol,whether or not subject to the pre-treatment temperature of 4°C or 10°C for 30 minutes.Analyses were performed immediately after freezing in liquid nitrogen (day 0) and after maintenance for 30, 60 and 90 days at -20°C.The viability was evaluated by traditional method of cultivation and production of transcripts by RT-PCR technique. The quantification was only possible on the first day of analysis (day 0) and had a mean of 3.0x107UFC and in the remaining periods of storage strains showed confluent growth not allowing their enumeration. The set of results has shown that the UHT milk was the most appropriate for cryopreservation than the use of neopeptona +12% glycerol. The pretreatment at 4°C for 30 minutes favored the production of transcripts for ciaB and dnaJ genes. For the strains ATCC 33291 and NCTC 11351 was verified a possible interconnection of sodB genes and p19, however, this link was not observed for the strain IAL 2383, which also showed different behavior from other strains for viability in both cryoprotectants and production of transcripts. The results of this study show that when the maintenance of viability of the strains is essential, it is necessary to use different combinations of cryoprotectants / treatments to increase the chances of recovery and, when the primary purpose is the production of transcripts, the option to maintain the reliability of the results is the immediate extraction of DNA of isolated strains.