Characterization of Wine Lactic Acid Bacteria: Single Broth Culture for Tests of Heterofermentation, Mannitol from Fructose, and Ammonia from Arginine

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Abstract
A single broth culture containing glucose, fructose, and L-arginine was used for testing three biochemical characteristics in strains of wine lactic acid bacteria (LAB). Heterofermentation was detected by observing gas formation from fermentation of the hexoses in inverted Durham tubes in the culture broths or by the dislodging of vaspar plugs over the culture broths, mannitol from fructose by typical rosette crystal formation upon drying of the broths, and ammonia from arginine by color development with Nessler's reagent. Both methods of gas detection resulted in 29 strains of 37 wine LAB tested being heterofermenters. The heterofermenters (24 Leuconostoc oenos and 5 Lactobacillus spp.) also produced mannitol from fructose, but the eight homofermenters (3 Lactobacillus spp and 5 Pediococcus spp) did not. Surprisingly, 75% of the leuconostocs produced ammonia from arginine, a characteristic typically absent from this genus (Bergey's Manual of Systematic Bacteriology, Vol 2, 1986). All of the heterofermentative lactobacilli were ammonia producers, but none of the homofermentative lactobacilli nor the homofermentative pediococci displayed this characteristic. The sensitivity of ammonia detection by Nessler's reagent was low. Use of higher levels (0.6%) of arginine in HFA broth compared with previous studies (0.3%) may be the reason for the detection of several strains of ammonia producing leuconostocs.

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