One loopful of each nodule suspension was aseptically streaked onto YEMA (10 g mannitol, 0.2 g MgS[O.sub.4]-7[H.sub.2]O, 0.2 g NaCl, 0.5 g [K.sub.2]HP[O.sub.4], 1 g yeast extract, and 15 g agar, pH 6.8 [+ or -] 0.2) medium containing bromothymol blue. The plates were wrapped with aluminium foil (for darkness), incubated at 28[degrees]C, and observed daily for the period necessary to characterize colony growth .
The formation of colonies on YEMA plates was monitored daily for 10 days, and the pH change of the growth medium was scored on YEMA plates containing 0.25 mg/l bromothymol blue (BTB).
Preparation of Bromothymol Blue
and [H.sub.2][O.sub.2] Solutions.
Rahman, "Optimized and validated spectrophotometric methods for the determination of amiodarone hydrochloride in commercial dosage forms using N-bromosuccinimide and bromothymol blue
," Journal of Saudi Chemical Society, vol.
Malic acid -- 5 g -- KOH -- 4 g -- [K.sub.2]HP[O.sub.4] 0.8 g -- 0.2 g K[H.sub.2]P[O.sub.4] 0.2 g 0.5 g 0.6 g MgS[O.sub.4].7[H.sub.2]O 0.2 g 0.01 g 0.2 g NaCl 0.2 g 0.02 g -- CaS[O.sub.4] 0.1 g -- -- Ca[Cl.sub.3] -- -- 0.02 g [H.sub.3]B[O.sub.3] 100 mg -- -- Fe[Cl.sub.3] -- -- 0.01 g ZnS[O.sub.4].7[H.sub.2]O 100 mg -- -- FeS[O.sub.4].7[H.sub.2]O 0.05 g -- MnS[O.sub.4].4[H.sub.2]O 10 mg 0.01 g -- CuS[O.sub.4].5[H.sub.2]O 3 mg -- -- [Na.sub.2]Mo[O.sub.4] -- 0.002 g 0.002 g Cane sugar -- -- 100 g KI 1 mg -- -- FeMo mixture 1 ml -- -- Bromothymol blue
(0.5% alc.soln) -- 2 ml 5 ml Distilled water 1000 ml 1000 ml 1000 ml Agar 20 g 1.75 g 20 g pH 7.3 6.6-7.0 6.0 FeMo mixture: Fe[Cl.sub.3].6[H.sub.2]O-14.5 g [Na.sub.2]Mo[O.sub.4].
Biochemical tests like congo red, bromothymol blue
(organic acid production test) and Gram reaction of isolates were carried out.
EXPERIMENT 2: TESTING FOR [CO.sub.2] With Ca[(OH).sub.2] or BROMOTHYMOL BLUE
Six native diazotrophic bacterial isolates isolates (ABA-1, ABA-10, ABA-14, ABA-2010, ACG-2 and ASA-1) and four reference strains (MTCC-446 and MTCC-124 (Azotobacter chroococcum), MTCC-1226 (Acetobacter diazotrophicus) and MTCC-2306(Azospirillum lipofarum) grown in respective nitrogen free media viz, Burk's medium for Azotobacter, diluted cane juice semi solid medium for Acetobacter and NFB (Nitrogen Free Bromothymol blue
medium) for Azospirillum (Cavalcant and Dobereiner, 1988) (Table-1).
and bromocresol purple as indicators of pH changes in chromatophores of Rhodospirillum rubrum.
were purified by dilution plating and identified by their morphologic and biochemical profiles, including development of blue color on L-canavanine-glycine bromothymol blue
Larvae were split opened with sterilized needles and scissors and a drop of the exuding haemolymph was streaked onto NBTA plates (nutrient agar (BDH) =37 g, Bromothymol blue
(Raymond) = 25 mg, 2, 3, 5- triphenyl-tetrazolium Chloride (1 %) (BDH) = 4 ml and distilled water = 1000 ml) with a sterilized inoculating loop.
These nematodes were surface sterilized, crushed individually in 100 [micro]L Luria broth with a motorized mortar and pestle, and plated on Luria broth and NBTA agar (nutrient agar supplemented with bromothymol blue
and triphenyltetrazolium chloride).