Keywords : Lactobacillus nagelii, wine, spoilage
INTRODUCTIONSluggish or stuck alcoholic fermentations are problems sometimes encountered by wine makers. These problem fermentations can be due to improper fermentation conditions or to insufficient nutrients being present in the grape must to support adequate yeast growth (Ough, 1966 ; Houtman et al., 1980a, b ;Ingledew & Kunkee, 1985 ;Kunkee, 1991). Recently, Huang et al. (1996) (Douglas & Cruess, 1936 ;Vaughn, 1955 ;Fornachon, 1957 ;Du Plessis & Van Zyl, 1963 ;Pilone et al., 1966 ; Chalfan et al., 1977 ;Maret & Sozzi, 1977, 1979Costello et al., 1983 Wibowo et al., 1985 ; Davis et al., 1986a, b ;Dicks & Van Vuuren, 1988 ;Sieiro et al., 1990). At the present time, L. kunkeei is the only species of Lactobacillus known that has been demonstrated to slow alcoholic fermentation of grape musts (Huang et al., 1996). However, most lactobacilli found in wines are considered to be spoilage organisms, due to production of acetic acid and\or other off-flavours (Davis et al., 1985). This study presents the biochemical characteristics and the results of phylogenetic analysis of a strain of Lactobacillus isolated from a partially fermented wine. On the basis of phenotypic and genotypic analysis of this micro-organism, a new species of Lactobacillus, Lactobacillus nagelii sp. nov., is proposed.
METHODSBacterial strains and cultivation. Strain LuE "! T was isolated from a commercial red wine obtained from L. Van Der Water (The Wine Lab, Napa, CA, USA). Control bacteria used during the biochemical characterization of LuE "! T were L. kunkeei ATCC 700308 and L. plantarum WS-16 (Edwards et al., 1993, 1998. All organisms were grown using modified Rogosa (MR) agar or broth supplemented with apple juice and adjusted to pH 4n5 (Beelman, 1982). Cultures were maintained on MR agar and in lyophilized form. C. G. Edwards and others washed twice in 5 ml phosphate buffer (pH 7, 0n023 M NaH # PO % \0n030 M Na # HPO % ), resuspended in 0n3 ml sterile phosphate buffer and inoculated into heterofermentationarginine broth described by Pilone et al. (1991). Tubes were overlaid with molten, sterilized vaspar (one part petroleum jelly and six parts paraffin) prior to incubation for 21 d at 25 mC. Production of ammonia from arginine, determination of optical isomers of lactic acid formed from glucose, dextran from sucrose and utilization of citric and malic acids were performed using the methods of Edwards et al. (1991). Nitrate reduction was tested as described by Carr (1970). Mannitol formation from fructose was demonstrated using the method of Pilone et al. (1991). Catalase was detected by placing drops of 3 % (w\v) H # O # on cultures growing on MR agar, altered by the addition of 0n5% (w\v) glucose, 20 % (v\v) apple juice and 0n0005 % (w\v) haematin and raising the pH from 4n5 to 5n5. Carbohydrate utilization was determined using the API Rapid CH system (bioMe! rieux) using the recommended CHL medium. API galleries were incubated for up to 21 d at 24-25 mC.For pH and temperature characteriz...