F. Ruiz-Berraquero

Numerical taxonomy of moderately halophilic Gram-negative rods

Summary: A study was made of 516 randomly selected isolates of moderately halophilic bacteria from solar salterns showing salinities between 8.8 and 40.0% (w/v) total salts, located in S.E. Spain. After purification, many cytological, physiological, biochemical, nutritional and antibiotic sensitivity characters were determined for 106 selected saltern isolates and two reference strains. Data were coded and analysed by numerical techniques using the Jaccard coefficient (S j), and clusters of strains were obtained by average linkage (UPGMA) analysis. Nine major phenons were found at the 72.5% similarity level. The properties of each phenon are given, their taxonomic affinities are discussed, and typical reference strains are suggested. Almost all the strains were related to genera known to contain marine species. A large number of the strains could be tentatively assigned to the genus Vibrio, suggesting that this may be an abundant taxon of moderately halophilic Gram-negative rods in solar salterns.

Deleya halophila, a New Species of Moderately Halophilic Bacteria

Deleya halophila sp. nov., which was isolated from hypersaline soils, is proposed. Each of 38 isolates, together with 8 reference strains, was examined in 97 phenotypic tests, and the data were analyzed by numerical taxonomy procedures. The 38 isolates formed a distinct group at a 76% similarity value, as determined by an analysis performed with the simple matching coefficient and unweighted average linkage clustering. Characteristically, the isolates were aerobic, gram-negative rods that were motile by one to eight peritrichous flagella. They grew optimally in the presence of 7.5% (wt/vol) marine salts. The distinguishing features of the new species are its salt requirement, biochemical features, and nutritional versatility. The guanine-plus-cytosine content of the deoxyribonucleic acid is 66.7 mol%. The type strain of this species is strain F5–7 (= CCM 3662).

Volcaniella eurihalina gen. nov., sp. nov., a moderately halophilic nonmotile gram-negative rod

A comparison of 16 gram-negative moderately halophilic aerobic rod-shaped bacteria with other halophilic and nonhalophilic gram-negative bacteria supported the establishment of Volcaniella eurihalina gen. nov., sp. nov. This comparison included phenotypic properties, salt requirements, and guanine-plus-cytosine contents of the DNAs, as well as DNA-DNA homology studies. The distinguishing features of this new bacterial genus are as follows: The organisms are nonmotile short rods that are oxidase negative; they are aerobic with a strictly respiratory type of metabolism; they are moderate halophiles, optimal growth occurs at a total salt concentration of 7.5% (wt/vol), and they exhibit a strongly euryhaline character; and they have a specific requirement for Na+ ions (sodium can be supplied as NaCl, Na2SO4, or NaBr). The minimum NaCl concentration required is 1.5% (wt/vol). The guanine-plus-cytosine content of the DNA is 59.1 to 65.7 mol%. This organism was isolated from hypersaline habitats, including saline soils and salt ponds, and from seawater. The type strain is strain F9-6 (= ATCC 49336).

Classification of “Chromobacterium marismortui” in a New Genus, Chromohalobacter gen. nov., as Chromohalobacter marismortui comb. nov., nom. rev.

Phenotypic and chemotaxonomic characteristics of seven moderately halophilic isolates from a mediterranean saltern showed that they were very closely related to “Chromobacterium marismortui.” This species, which was originally described on the basis of strains isolated from the Dead Sea, was not included on the Approved Lists of Bacterial Names and is not accepted as a member of the genus Chromobacterium since it produces a pigment that is not violacein, does not possess the characteristic flagellar arrangement, and is not genotypically related to this genus as determined by deoxyribonucleic acid-ribosomal ribonucleic acid hybridization experiments. On the basis of the special features of these strains, we propose that they should be placed in a new genus, Chromohalobacter, which includes a single species, Chromohalobacter marismortui comb. nov., nom. rev. The deoxyribonucleic acid base composition is 62.1 to 64.9 mol% guanine plus cytosine. The type strain is strain ATCC 17056 (= CCM 3518).

SHORT COMMUNICATION Isolation of Extremely Halophilic Bacteria Able to Grow in Defined Inorganic Media with Single Carbon Sources

Summary: Pigmented pleomorphic rods morphologically similar to Halobacterium volcanii were isolated from a salt pond near Alicante, Spain. The salt concentration range for growth (10 to 35 %, w/v, total salts) was more moderate than that typical of Halobacterium. Twenty-two strains were characterized as members of the genus Halobacterium, but were distinguished from currently recognized species by the absence of complex nutrient requirements, a rapid growth rate, nutritional versatility and salt-dependent pigment production. A typical strain (R-4) has been deposited in the Czechoslovak Collection of Microorganisms, no. CCM 3361.

Taxonomic Study and Amended Description of Vibrio costicola

A total of 54 moderately halophilic vibrios, which were isolated from several salterns located in different areas of Spain, were examined by using a wide range of morphological, physiological, biochemical, and nutritional tests. The resulting data, together with data for four reference Vibrio costicola strains including the type strain V. costicola NCMB 701, and other marine species that were similarly examined, were compared by using several numerical taxonomic methods. There was a strong similarity between the 54 isolates and four reference strains of V. costicola that were isolated from cured meats. On the basis of these and other molecular data, including guanine-plus-cytosine content of the deoxyribonucleic acid and the plasmid content, we propose an amended description of this species.

Salinicoccus roseus gen. nov., sp. nov., a New Moderately Halophilic Gram-Positive Coccus

Summary A new moderately halophilic Gram-positive coccus isolated from a solar saltern located in Alicante (Spain) is described. The organism required media with NaCl concentrations between 0.9 and 25% (w/v), with the optimum at 10% (w/v) NaCl. It was non-motile, non-sporing, oxidase and catalase positive. The GC content of its DNA was 51.2 mol%. The murein was of the L-Lys-Gly 5 type. The organism is different from previously described cocci. The name Salinicoccus roseus gen. nov., sp. nov. is proposed. The type strain is strain no. 9 which has been deposited as ATCC 49258, DSM 5351, and CCM 3516.

Bacillus halophilus sp. nov., a Moderately Halophilic Bacillus Species

Summary Bacillus halophilus, a new moderately halophilic (optimum growth at 15% salt) species, is described. This species was isolated from rotting wood from the Pacific Ocean. The G+C mol % (Tm) of its deoxyribonucleic acid is 51.5. The type strain is N23-2 (= ATCC 49085 = DSM 4771 = CCM 4074).

Deleya salina sp. nov., a Moderately Halophilic Gram-Negative Bacterium

Deleya salina sp. nov., which was isolated from hypersaline habitats, is described. Our description is based on 26 strains that exhibit high levels of phenotypic similarity (simple matching coefficient, >74%) and DNA-DNA homology (<71%). The organisms are moderately halophilic gram-negative rods; growth occurs at salt concentrations between 2.5 and 20% (wt/vol) (optimal growth occurs at a salt concentration of 5% [wt/vol]). D. salina is aerobic, nonmotile, and catalase and oxidase positive. The cells accumulate poly-β-hydroxybutyrate, and the G+C contents of the DNAs range from 60.4 to 64.2 mol%. The type strain is strain F8-11 (= ATCC 49509).

Marinococcus hispanicus, a New Species of Moderately Halophilic Gram-Positive Cocci

Five moderately halophilic gram-positive cocci were isolated from soils and ponds of a solar saltern located near Alicante, Spain. These organisms all were nonmotile, nonsporing, reddish orange pigmented, strictly aerobic, and catalase and oxidase positive, had meso-diaminopimelic acid in their cell walls, and had DNA base compositions ranging from 45.7 to 49.3 mol%; they constitute a homology group with levels of DNA-DNA homology ranging from 70 to 100%. On the basis of their features, we regard these strains as belonging to a new species of the genus Marinococcus, for which we propose the name Marinococcus hispanicus. The type strain is strain J-82 (= ATCC 49259 = DSM 5352 = CCM 4148).