No abstract

One hundred and seventy-eight lactobacilli isolated from wine were characterized by a polyphasic approach. Strains were phenotypically identified at genus and species level by classical tests including the analysis of cell morphology, homo/heterofermentative character, sugar fermentation patterns, growth at different temperatures and the optical nature of the isomer of lactic acid produced from glucose. Molecular techniques such as random amplification of polymorphic DNA (RAPD), amplified 16S rDNA restriction analysis (16S-ARDRA), PFGE-RFLP and ribotyping were used to characterize strains, and their potential for identification and/or typing was evaluated. The information obtained with these techniques was processed with the BioNumerics software in order to analyse relationships existing between isolated strains and various reference species of the genus. Then, taxonomic dendrograms were obtained, and this information allowed the proposal of molecular procedures suitable for the identification and typing of these wine micro-organisms. The techniques useful for both identification and typing were RAPD and ribotyping, while 16S-ARDRA was only useful for identification and PFGE-RFLP only for typing purposes. The wine strains were identified as Lactobacillus brevis (19 strains), Lactobacillus collinoides (2 strains), Lactobacillus hilgardii (71 strains), Lactobacillus paracasei (13 strains), Lactobacillus pentosus (2 strains), Lactobacillus plantarum (34 strains) and Lactobacillus mali (10 strains).

Six strains with more than 99?5 % 16S rRNA gene sequence similarity, identical internal spacer region profiles and restriction analysis of the amplified 16S rRNA gene patterns were isolated from fermenting grape musts during independent studies carried out in France and Spain many years apart. Strains are Gram-positive, motile, facultatively anaerobic rods that do not exhibit catalase activity and have the ability to utilize pentose sugars (ribose and/or L-arabinose), although they are homofermentative bacteria. Strains ferment pentoses exclusively yielding lactic acid as the end product. A broad set of molecular techniques has been applied to characterize these strains and the results show a high degree of genotypical congruence, sharing identical profiles with 16S rRNA-based techniques. Phylogenetic analysis based on 16S rRNA gene sequences placed these strains within the genus Lactobacillus, closely related to Lactobacillus mali, Lactobacillus nagelii and Lactobacillus satsumensis (with approximately 95 % sequence similarity). DNA-DNA hybridization experiments confirmed the independent status at the species level of these fermenting grape-musts strains. Phenotypically they can be distinguished from the closest relatives by several traits such as growth temperatures and fermentation of carbohydrates. The name Lactobacillus vini sp. nov. is proposed, with strain Mont 4 T (=DSM 20605 T =CECT 5924 T ) as the type strain.Several species of lactic acid bacteria have been described by different authors as being associated with the winemaking process Wibowo et al., 1985;Fleet, 1993;Rodas et al., 2005). Some of them are beneficial for wine quality, whereas others have been described as detrimental (Sponholz, 1993;Edwards et al., 1998).In 1978, P. Barre isolated one wine lactobacillus from hightemperature fermenting grape must (Barre, 1978), named as Mont 4, that was deposited at the Deustche Sammlung von Micro-organismem und Zellkulturen GmbH (DSMZ) as Lactobacillus sp. DSM 20605. Afterwards, this bacterium was incorporated in the Spanish Type Culture Collection (CECT) from DSMZ as Lactobacillus sp. CECT 5924. Chenoll et al. (2003) included it in a molecular taxonomic study in order to establish an rRNA-based identification scheme for Carnobacterium, Lactobacillus, Leuconostoc and Pediococcus species. In this study, the authors reported that strain CECT 5924 (=Mont 4=DSM 20605) had singular rRNA-RFLP, internal spacer region (ISR)-PCR and ISR-RFLP profiles that allowed its differentiation from the rest of the analysed members of the genus Lactobacillus. In addition, and based on 16S rRNA gene sequence analysis, it was suggested that this strain might represent a novel species. Later on, Rodas et al. (2005), following an extensive polyphasic study of wine lactobacillus strains, found a cluster of wine lactobacilli that showed 100 % 16S rRNA gene sequence similarity value with strain Mont 4 (=CECT 5924). The closest phylogenetic neighbours of strain CECT 5924 are Lactobacillus mali, Lactobacillus nagelii and the recen...

A Lactobacillus strain, designated 203 T , previously isolated from Bobal grape must was characterized phylogenetically, genotypically and phenotypically in order to establish whether it represents a novel species. On the basis of the 16S rRNA gene sequence, strain 203 T was shown to belong to the genus Lactobacillus, falling within the Lactobacillus alimentariusLactobacillus farciminis group and being closely related to the type strains of L. alimentarius, Lactobacillus kimchii and Lactobacillus paralimentarius. DNA-DNA hybridization results confirmed the separate status of strain 203 T at the species level. To establish the similarities and differences between 203 T and the three aforementioned closest species, the following methods were used: amplified rDNA restriction analysis, analysis of the 16S-23S rDNA intergenic spacer region, random amplification of polymorphic DNA (RAPD) profiling, ribotyping, carbohydrate fermentation and physiological tests. Strain 203 T could be differentiated genetically using RAPD analysis and ribotyping. Phenotypically, it can be distinguished from its closest relatives by its ability to grow at pH 3.3, by gas production from gluconate and by certain carbohydrate fermentations. On the basis of these data, strain 203 T represents a novel species of the genus Lactobacillus, for which the name Lactobacillus bobalius sp. nov. is proposed. The type strain is 203 T (5CECT 7310 T 5DSM 19674 T ).Eighteen different species of the genus Lactobacillus have been reported to be involved in the winemaking process (Chalfan et al., 1977;Costello et al., 1983;Davis et al., 1986;Lafon-Lafourcade, 1983;Lafon-Lafourcade et al., 1983;Maret & Sozzi, 1977, 1979 Ribéreau-Gayon et al., 2000b;Rodas et al., 2005;Wibowo et al., 1985). In recent years, three of them have been described as novel species, namely Lactobacillus kunkeei (Edwards et al., 1998), Lactobacillus nagelii (Edwards et al., 2000) and Lactobacillus vini (Rodas et al., 2006). Wine lactobacilli are sometimes considered as spoilage organisms since they can produce acetic acid and/ or off-flavours Wibowo et al., 1985;Fleet, 1993;Sponholz, 1993) and biogenic amines (Lonvaud Funel, 2001; Ribéreau-Gayon et al., 2000a;Landete et al., 2005).In an earlier study, Rodas et al. (2005) carried out an extensive polyphasic analysis using genotypic and phenotypic methods to characterize 170 lactobacilli isolated from musts and wines. The dendrogram built by comparing the results obtained from the different techniques showed that strain 203 T did not cluster with any of the reference species of the genus Lactobacillus. An analysis of 16S rRNA gene sequence similarity showed that strain 203 T was most closely related to the type strains of Lactobacillus kimchii (99.3 %), Lactobacillus paralimentarius (97.8 %) and Lactobacillus alimentarius (98.2 %). However, the study results did not confirm whether strain 203 T represented a novel species or not.The phylogenetic, genotypic and phenotypic characterization of isolate 203 T and its closest phylogenetic relati...

The study is the first report on the range of supply reduction measures in Australian prisons and, possibly in the world. Both measures were employed extensively across the country and finds of drugs and drug use were relatively low. Two possible conclusions can be drawn; that either drug use was very low in prison or that it was well concealed from the authorities. A comparison of random testing with targeted testing of inmates, where the former yields fewer positive results shows drug use was likely to be low rather than well concealed.