Maintenance of strains of Campylobacter jejuni in laboratories after use of cryoprotectors and pre-treatment of stress

Roberta Torres de Melo, Mariela Moura Carreon, Guilherme Paz Monteiro, Eliane Pereira Mendonça, Phelipe Augusto Borba Martins Peres, Raquelline Figueiredo Braz, Fernanda Aparecida Longato dos Santos, Daise Aparecida Rossi

Abstract


This study evaluated the feasibility and the production of transcripts of sodB, p19, ciaB and dnaJ genes in strains of Campylobacter jejuni ATCC 33291, NCTC 11351, and 2383 IAL stored in whole UHT milk and or neopepton + 12% glycerol, submitted or not to pre-treatments at 4°C or 10°C for 30 minutes. The analyzes were performed immediately after freezing in liquid nitrogen (day 0) and after maintenance for 30, 60, and 90 days at -20ºC. The viability was evaluated by the traditional culture method and the production of transcripts by the RT-PCR technique. The quantification was only possible on the first day of analysis (day 0) and presented a mean of 3.0 x 107 CFU, and in the other periods of storage the strains presented confluent growth, not allowing their enumeration. The results indicated that whole UHT milk was more adequate for cryopreservation than the use of neopepton + 12% glycerol. The use of pre-treatments combined with the use of UHT milk as a cryoprotective medium stabilized the cells in order to transcribe the ciaB, dnaJ, sodBand p19 genes in the strains maintained under -20 ° for 30 to 60 days, indicating that they are more suitable methods for the maintenance of strains in the laboratory.

Keywords


Campylobacteriosis; Cryopreservation; RT-PCR; Virulence.

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References


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DOI: http://dx.doi.org/10.5433/1679-0359.2019v40n6Supl2p3305

Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433/1679-0359
E-mail: semina.agrarias@uel.br
Este obra está licenciado com uma Licença Creative Commons Atribuição-NãoComercial 4.0 Internacional