Growth and Characterization of G. anatis Strains

The G. anatis strains used in this work were 12656-12 (β-hemolytic biovar, isolated from chicken liver by Bojesen et al., 2003 (link)) and ESV200 (β-hemolytic, isolated in Tepatitlan, Jalisco, Mexico (geographical coordinates: 20° 49′ 0″ north, 102° 44′ 0″ west), in 2009 from hens with respiratory disease by Dr. Edgardo Soriano). Strain purity was checked by streaking on blood agar medium with incubation at 37°C for 24 h. For growth in liquid medium, strains were grown at 37°C for 24 h in brain-heart infusion (BHI) medium (Merck®) with aeration, unless specified otherwise. Precultures were incubated until the optical density at 595 nm (OD₅₉₅) was 1.0 before inoculation in BHI broth in either flasks or microplates (Chantes-Guerra et al., 2022 (link)).