F. Gosselé

Frateuria, a New Genus for “Acetobacter aurantius”

The properties and taxonomic positions of 11 strains previously identified as members of “Acetobacter aurantius Kondo and Ameyama” (this name is not on the Approved Lists) were reexamined. For each we determined about 100 phenotypic features, the protein gel electropherograms, and the parameters of deoxyribonucleic acid:14C-ribosomal ribonucleic acid (DNA:[14C]rRNA) hybrids. The 11 strains fell into three taxonomically distinct groups. Strain IFO 3248 was the only one which belonged in Acetobacter. Strain IFO 3246 was an atypical Gluconobacter. The remaining nine strains formed a tight cluster, with very similar phenotypic features and protein gel electropherograms. Taxonomically, this cluster is quite removed from Gluconobacter and Acetobacter, and the properties of its DNA:rRNA hybrids suggest that it is closer to Pseudomonas Section I (R. E. Buchanan and N. E. Gibbons [ed.], Bergeys Manual of Determinative Bacteriology, 8th ed.) and Xanthomonas. We propose the name Frateuria gen. nov. for this cluster, with Frateuria aurantia sp. nov. as the type species and IFO 3245 as the type strain. An extensive phenotypic description and minimal standards of the new genus are given, as is the phenotypic differentiation from Gluconobacter and Acetobacter.

Numerical Analysis of Phenotypic Features and Protein Gel Electropherograms of Gluconobacter Asai 1935 emend. mut. char. Asai, Iizuka, and Komagata 1964

Ninety-eight Gluconobacter strains of various origins were examined by numerical analysis of 177 phenotypic features and by gel electrophoresis of soluble proteins. Gluconobacter was phenotypically quite different from Acetobacter and Frateuria. An extensive phenotypic description and a minimal description of the genus Gluconobacter are given. The genus Gluconobacter contained two groups, A and B, by both techniques. Phenons A and B could be differentiated only by the requirement for nicotinic acid and by their electrophoretic patterns. Protein electrophoresis showed clearly that Gluconobacter strains are genetically stable over several decades. The strains of all five subspecies of Gluconobacter oxydans cited in the Approved Lists of Bacterial Names (Skerman et al., Int. J. Syst. Bacteriol. 30:225–420, 1980) were distributed randomly over the phenotypic and electrophoretic clusters and subclusters, and the type strains of the subspecies all fell in phenon B. We conclude that the single species Gluconobacter oxydans should not be further divided into subspecies.

Numerical Analysis of Phenotypic Features and Protein Gel Electrophoregrams of a Wide Variety of Acetobacter strains. Proposal for the Improvement of the Taxonomy of the Genus Acetobacter Beijerinck 1898, 215

Ninety-eight strains, representing all Acetobacter species and subspecies from the Approved Lists of Bacterial Names (Skerman et al., 1980), were examined in a numerical analysis of 177 phenotypic features and compared to ninety-eight Gluconobacter and seven Frateuria strains. Four phenons could be delineated, corresponding to Frateuria (phenon 1), A. aceti subsp. liquefaciens (phenon 2), Gluconobacter (phenon 3) and Acetobacter minus A. aceti subsp. liquefaciens (phenon 4). Acetobacter, Frateuria and Gluconobacter are well- could be distinguished. Comparison of the protein electrophoregrams of Acetobacter strains revealed a fairly high internal homogeneity within phenon 2, subphenons C and D. Strains of the subphenon E gave very divergent protein patterns. The following classificatory changes are proposed within the genus Acetobacter: (1) Acetobacter liquefaciens sp. nov. is proposed for the homogeneous phenon 2, containing all 12 A. aceti subsp. liquefaciens strains (% G + C range of 62.3 to 64.6; IAM 1834 as type strain); (2) for the homogeneous subphenon D containing 8 A. aceti subsp. aceti strains, the name Acetobacter aceti emend, should be retained (% G + C range of 55.9 to 59.5; NCIB 8621 as type strain); (3) for subphenon E, a heterogeneous group, containing a variety of Acetobacter subspecies (all with their type strain) the species name Acetobacter pasteurianus emend, is preserved with LMD 22.1 as type strain; this species has the broad % G + C range of 52.8 to 62.5; (4) for subphenon C, a new species, Acetobacter hansenii sp. nov. is proposed (% G + C range of 58.1 to 62.6, NCIB 8746 as type strain). Minimal descriptions and differentiating keys are provided.

A rapid, simple and simultaneous detection of 2-keto-, 5-keto-and 2,5-diketogluconic acids by thin-layer chromatography in culture media of acetic acid bacteria

Summary A rapid and simple test for the simultaneous detection of 2-keto-, 5-keto- and 2.5-diketo-gluconic acids by thin-layer chromatography and their revelation with o-phenylene-diamine. HCl is described. 32 Acetobacter and 83 Gluconobacter strains were scanned with this method. The formation of keto-gluconic acids appears to be a stable and reliable feature for the classification of acetic accid bacteria.

The bacterial microflora of witloof chicory (Cichorium intybus L. var.foliosum Hegi) leaves.

The bacterial flora on the heads of four different witloof chicory varieties was examined. The 590 isolates were characterized by their SDS-PAGE protein profiles; they revealed 149 different protein fingerprint types. The fluorescentPseudomonas fingerprint type CH001 was abundantly found on all heads examined. Fourteen other fingerprint types occurred in high densities more than twice. Among these, the following were identified: fluorescentPseudomonas, nonfluorescentPseudomonas sp.,Erwinia herbicola, Erwinia sp., andFlavobacterium sp. The majority of the fingerprint types (90%) was found only once. It was also our objective to isolate bacteria applicable in the biological control of chicory phytopathogens. Isolates of all fingerprint types were tested for in vitro antagonistic activity and for possible deleterious effect on plant growth. FluorescentPseudomonas andSerratia liquefaciens isolates were antagonistic against fungi. Among the 161 fluorescentPseudomonas strains, five were able to produce disease symptoms on chicory leaves upon inoculation. Comparison of the results of this study with those obtained in two previous analyses revealed that the leaf microflora showed some similarities with the bacterial flora of chicory roots. The chicory seed microflora differed from that of both leaves and roots.

Gluconobacters from honey bees

Fifty-sixGluconobacter strains and oneAcetobacter strain were isolated from honey bees and their environment in three different regions in Belgium and identified phenotypically.Polyacrylamide gel electrophoresis of the soluble cell proteins showed that two different types exist within theGluconobacter isolates: strains from type A were found in samples of the three regions, whereas strains from type B were only isolated in two of the three regions. Both types could occur in bees from the same region, from several hives of one bee keeper and from one hive. Strains from type A were almost identical with collection strainG. oxydans subsp.suboxydans NCIB 9018, whereas strains from type B consituted a new protein electrophoretic type within the genusGluconobacter.AlthoughGluconobacter is apparently associated with honey bees, it is not known whether it is important or required for the bees or any hive product.

The nitrogen requirements ofGluconobacter, Acetobacter andFrateuria

The nitrogen requirements of 96Gluconobacter, 55Acetobacter and 7Frateuria strains were examined. Only someFrateuria strains were able to grow on 0.5% yeast extract broth or 0.5% peptone broth. In the presence ofd-glucose ord-mannitol as a carbon source, ammonium was used as the sole source of nitrogen by all three genera. With ethanol, only a fewAcetobacter strains grew on ammonium as a sole nitrogen source. Singlel-amino acids cannot serve as a sole source of carbon and nitrogen for growth ofGluconobacter, Acetobacter orFrateuria. The singlel-amino acids which were used by most strains as a sole nitrogen source for growth are: asparagine, aspartic acid, glutamine, glutamic acid, proline and alanine. SomeAcetobacter andGluconobacter strains deaminated alanine, asparagine, glutamic acid, threonine, serine and proline. NoFrateuria strain was able to develop on cysteine, glycine, threonine or tryptophan as a sole source of nitrogen for growth. An inhibitory effect of valine may explain the absence of growth on this amino acid. No amino acid is “essential” forGluconobacter, Acetobacter orFrateuria.

Identification of Acetobacter strains isolated from spoiled lactic acid fermented meat food for pets.

FiveAcetobacter isolates from lactic acid fermented meat food for pets were characterized by 177 morphological, physiological and biochemical traits. Four isolates were identified asA. pasteurianus, one asA. aceti. It is emphasized that access of such bacteria to lactic acid fermented foods should be avoided.

Rediscovery of Bertrand's Sorbose Bacterium (Acetobacter aceti subsp. xylinum): Proposal to Designate NCIB 11664 in Place of NCIB 4112 (ATCC 23767) as the Type Strain of Acetobacter aceti subsp. xylinum: Request for an Opinion

None of three culture collection strains (NCIB 4112, ATCC 23767, and CIP 57.14) presumed to be Bertrands sorbose bacterium (type strain of Acetobacter aceti subsp. xylinum) was found to be a member of the genus Acetobacter. These three strains were identical, displayed considerable phenotypic differences compared with descriptions of the original sorbose bacterium published before 1959, and belonged to Gluconobacter oxydans. However, a 1949 Sordelli-dried culture of strain NCIB 4112 was revived at the National Collection of Industrial Bacteria and was found to belong to the genus Acetobacter. This culture showed similarities to other A. aceti subsp. xylinum strains and therefore is most similar to the culture originally deposited in 1933 by Bertrand in the National Collection of Type Cultures. The new accession number of the revived sorbose bacterium is NCIB 11664, and we request an opinion to designate this strain as the type strain of A. aceti subsp. xylinum in place of strain NCIB 4112.

Growth factor requirements of Gluconobacter

Summary The growth factor requirements of ninety-five Gluconobacter strains are examined: 58% of them require pantothenic acid only, 28% require pantothenic acid and nicotinic acid and 6% require pantothenic acid, nicotinic acid and thiamin. Growth factor requirements are strain-specific and are not correlated with the present subdivisions within the genus.