Tryptic soy agar tsa

All microorganisms used in this study are listed in Table 1. Cell stocks were kept at −80 °C and subcultured once before the experiments. Filamentous fungi strains were maintained in Sabouraud Dextrose Agar (SDA) (Liofilchem s.r.l., Roseto D.A., Italy) or Potato Dextrose Agar (PDA) (Liofilchem s.r.l., Roseto D.A., Italy) for approximately 7 days at room temperature. For each experiment, conidia were harvested by flooding the agar surface with sterilized saline solution containing NaCl 8.00 g·L⁻¹ (Sigma-Aldrich, Sintra, Portugal), KCl 0.2 g·L⁻¹, Na₂HPO₄·2H₂O 1.44 g·L⁻¹, and KH₂PO₄ 0.24g·L⁻¹ (all from José Manuel Vaz Pereira, Benavente, Portugal) (pH 7.4). Biomass was then suspended in the saline solution with a sterile loop and the final solution collected with a pipette tip to a sterile tube. The heavier fragments were allowed to deposit in the bottom for 5–10 min and subsequently the supernatant was transferred to a new sterile tube [26 (link),27 (link)]. Candida strains were maintained on Sabouraud dextrose agar for 48 h at 37 °C [28 (link)]. Bacterial strains were maintained on Tryptic soy agar (TSA) (Liofilchem s.r.l., Roseto D.A., Italy) for 24 h at 37 °C [29 (link)]. Strains were provided by Colección Española de Cultivos Tipo (CECT), Micoteca da Universidade do Minho (MUM), and the American Type Culture Collection (ATCC).

Compounds/extracts were tested against the following bacterial strains: Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213, Staphylococcus epidermidis ATCC 14990, Streptococcus pyogenes ATCC 19615, and Micrococcus luteus ATCC 4698. Prior to each in vitro bioassay, fresh cultures were obtained for each strain using the appropriate medium and incubation conditions as follows. Staphylococcus spp. and P. aeruginosa were grown on Mueller–Hinton (MH) agar (Liofilchem srl, Roseto degli Abruzzi, Italy) for 24 h at 37 °C, while M. luteus was grown in Tryptic Soy agar (TSA, Liofilchem srl, Italy) for 24 h at 30 °C and S. pyogenes on TSA supplemented with 5% defibrinated sheep blood (Thermo Fisher Scientific, Waltham, MA, USA) for 24 h at 37 °C in an atmosphere of 5% CO₂.

Forty-nine Aeromonas strains were used in this study: 46 Georgian strains, majority collected from sick fish and water in 5 fish farms in Central Georgia, including 39 strains of A. hydrophila, 5 strains of A. caviae, and 2 strains A. sobria (collection of the Eliava Institute. 3 Gotua street, 0160 Tbilisi, Georgia); 3 reference strains: A. hydrophila CIP103770, A. salmonicida achromogenes CIP 104001T, and A. salmonicida salmonicida CIP 103209T, obtained from Collection of Institute Pasteur (Collection De L’Institut Pasteur (CIP), 28 rue du Docteur Roux 75724 Paris CEDEX 15). For the detailed list of strains, see Tables S1 and S2.
For propagation of bacterial strains and for testing of phage activity, Tryptic Soy Broth (TSB) and Tryptic Soy Agar (TSA) (Liofilchem, Italy) were used. The characteristic cultural properties of Aeromonas strains were checked on Aeromonas Selective Agar (ASA) (Liofilchem, Italy).