P. salmonis LF-89T (ATCC VR-1361) type strain and the Chilean isolates LF-89-like 1, EM-90-like 1, and EM-90-like 2 were routinely grown as a bacterial lawn on BBL™ Trypticase Soy Agar (Becton, Dickinson and Company, Sparks, MD, USA) plates supplemented with NaCl 15 g/L, fetal bovine serum (FBS) 5%, l- cysteine 0.1%, d- glucose 0.5%, and FeCl3 0.120 mM at 18 °C for five days. After that, P. salmonis were carefully scraped from bacterial lawn and suspended onto 5 mL of Tryptic Soy Broth (TSB) (Merck, Darmstadt, Germany) supplemented with NaCl 3 g/L, FBS 2.5%, l -cysteine 0.05%, and FeCl3 0.01 g/L and grown at 18 °C and 100 rpm for two days [31 (link)] prior to the biofilm formation assays. The identified strains were confirmed by PCR assays [30 (link)] and 16S rRNA sequencing [32 (link)].
Bbl trypticase soy agar
Manufactured by BD
Sourced in United States
BBL™ Trypticase Soy Agar is a general-purpose microbiological growth medium. It is designed to support the growth of a wide range of bacterial species.
Automatically generated - may contain errors
Lab products found in correlation
Tryptic soy broth (tsb), by Merck Group (1 mentions)
Xanthine, by Merck Group (1 mentions)
Rra pt va, by Promega (1 mentions)
Phoenix m50, by BD (1 mentions)
Maldi tof ms, by Bruker (1 mentions)
Yeast extract, by Thermo Fisher Scientific (1 mentions)
Du650 spectrophotometer, by Beckman Coulter (1 mentions)
4 protocols using bbl trypticase soy agar
1
Cultivation and Identification of Piscirickettsia salmonis
2
Metal-Dependent Enzyme Assay Protocols
3
Identification and Antibiotic Susceptibility Testing
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Positive samples were subcultured on the BAP/EMB bi-plate (BBL™Trypticase™ Soy Agar with 5% Sheep Blood/Levine EMB Agar, BD) and further incubated overnight at 35 °C with CO2. The bacterial colonies were subsequently picked for ID by MALDI-TOF MS (Bruker Daltonik, Bremem, Germany) and followed by AST with a BD Phoenix™ M50. When comparing the peptide mass fingerprint of unknown samples to the Bruker Biotyper reference database (Bruker, Billerica, MA, USA), a similarity score > 1.7 [percentage of reliable ID (%)] indicates a match to the optimal genus-level, thereby rendering practical indication for clinical treatment.
4
Cultivation of A. actinomycetemcomitans Strains
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A.actinomycetemcomitans strains, D7S-SA (wild type Aa) and D7S-SA CHE001 (Cdt deficient Aa mutant), were obtained as described by Nalbant, A., et al (Nalbant et al., 2003 (link)). Each strain was plated on AAGM agar which consisted of 20g of BBL trypticase soy agar (BD; Sparks, MD); 3g of yeast extract (ThermoFisher) supplemented with 0.4% sodium bicarbonate and 0.8% dextrose (Sreenivasan et al., 1993 (link)). After bacteria were grown on plates for 24 or 48 hours in the incubator with 10% CO2 at 37°C, they were inoculated in 10ml of AAGM broth until OD600 close to 0.2. Bacteria were plated from different dilutions at various time points on AAGM agar plates and incubated for 24 hours in the incubator with 10% CO2 at 37°C (Nalbant et al., 2003 (link)). OD600 was measured using DU 650 Spectrophotometer (Beckman Coulter, Indianapolis IN) for each time point.
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A.actinomycetemcomitans strains, D7S-SA (wild type Aa) and D7S-SA CHE001 (Cdt deficient Aa mutant), were obtained as described by Nalbant, A., et al (Nalbant et al., 2003 (link)). Each strain was plated on AAGM agar which consisted of 20g of BBL trypticase soy agar (BD; Sparks, MD); 3g of yeast extract (ThermoFisher) supplemented with 0.4% sodium bicarbonate and 0.8% dextrose (Sreenivasan et al., 1993 (link)). After bacteria were grown on plates for 24 or 48 hours in the incubator with 10% CO2 at 37°C, they were inoculated in 10ml of AAGM broth until OD600 close to 0.2. Bacteria were plated from different dilutions at various time points on AAGM agar plates and incubated for 24 hours in the incubator with 10% CO2 at 37°C (Nalbant et al., 2003 (link)). OD600 was measured using DU 650 Spectrophotometer (Beckman Coulter, Indianapolis IN) for each time point.
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