Newly Cultured Bacteria with Broad Diversity Isolated from Eight-Week Continuous Culture Enrichments of Cow Feces on Complex Polysaccharides

↵a All chemicals were purchased from Sigma.

↵b Mineral 1 is K₂HPO₄, 6 g liter⁻¹.

↵c Mineral 2 is KH₂PO₄, 6 g liter⁻¹; (NH₄)₂SO₄, 6 g liter⁻¹; NaCl, 12 g liter⁻¹; MgSO₄, 2.45 g liter⁻¹; and CaCl₂ · 2H₂O, 1.69 g liter⁻¹.

↵d Added as a solution of 0.5 g hemin in 25 ml 1 M NaOH.

↵e Short chain fatty acid solution contained the following acids, in ml/liter: acetic, 6.85; propionic, 3.00; butyric, 1.85; isobutyric, 0.50; 2-methyl butyric, 0.55; N-valeric, 0.55; and isovaleric, 0.55.

↵f Two hundred milliliters anaerobic solution at pH 10 with 2.5 g l-cysteine HCl · H₂O and 2.5 g Na₂S · 9H₂O.

↵g Incubated clarified rumen fluid modified from the work of Allison et al. (50) as follows: 400 ml rumen fluid with addition of 300 ml mineral 1, 300 ml mineral 2, and 4 g Na₂CO₃; addition of a second centrifugation after overnight storage at 4°C; and adjustment to pH 7.0 prior to autoclaving. The fluid was stored frozen at −20°C until it was used.