- Volume 36, Issue 2, 1986

The name Providencia heimbachae sp. nov. is proposed for a group of organisms that were isolated from the feces of penguins in the Federal Republic of Germany and from a cow in the United States. P. heimbachae strains are gram-negative, oxidase-negative, fermentative, rod-shaped organisms that grow on MacConkey agar, as well as other media that are selective for members of the Enterobacteriaceae. The 13 P. heimbachae isolates which we studied gave negative results for the following tests: indole production, Voges-Proskauer, Simmons citrate, H2S production on triple sugar iron and Kligler agars, urease, lysine decarboxylase, arginine dihydrolase, gelatinase, growth in the presence of KCN, malonate, acid production from l-arabinose, cellobiose, dulcitol, erythritol, lactose, melezitose, melibiose, a-methylglucoside, raffinose, salicin, d-sorbitol, l-sorbose, starch, d-tagatose, trehalose, d-turanose, and d-xylose (15% positive after 7 days), esculin hydrolysis, acetate, lipase, deoxyribonuclease, and o-nitrophenyl-β-D-galactopyranoside. The strains were positive for the following tests: catalase, nitrate reductase, methy1 red, phenylalanine deaminase, acid production from d-glucose, adonitol, d-arabitol, d-fructose, d-galactose, d-mannose, and l-rhamnose, and tyrosine clearing. Acid production from glycerol, myo-inositol, maltose, and d-mannitol was delayed; gas was produced from d-glucose, d-galactose, and d-mannose in small amounts or not at all (54% positive after 7 days). Motility at 36°C was variable (46% positive after 2 days; 85% positive after 7 days). Deoxyribonucleic acids from 12 other strains of P. heimbachae were highly related (91 to 100% related in reactions assayed on hydroxyapatite at 60 and 75°C) to 32P-labeled deoxyribonucleic acid from the proposed type strain (strain ATCC 35613). Labeled deoxyribonucleic acid from this type strain was 22 to 45% related at 60°C and 4 to 31% related at 75°C to deoxyribonucleic acids from four other Providencia species. The levels of relatedness of P. heimbachae to Proteus species ranged from 10 to 13% in 60°C reactions. P. heimbachae could be differentiated from the other Providencia species by the following characteristics: negative tests for Simmons citrate, urease, indole production, and acid production from trehalose and positive tests for acid production from adonitol, d-arabitol, d-galactose, and l-rhamnose. All of the P. heimbachae strains which we studied were resistant to tetracycline, and most strains were resistant to cephalothin.

The phenetic taxonomy of 305 strains of Pseudomonas and related organisms was numerically studied by using 215 features, including 156 assimilation tests. A total of 200 field strains were isolated from spoiling meat, and 50 strains were isolated from freshwater or soil. In addition, 55 reference strains (including 23 type strains and 4 clinical strains) were obtained. The strains clustered into 25 clusters at the 75% level when the Jaccard similarity coefficient was used. The 10 clusters that were considered significant were assigned to the Pseudomonas fragi complex (131 strains), Pseudomonas lundensis (40 strains), Pseudomonas fluorescens biovar 1 (27 strains), P. fluorescens biovar 2 (5 strains), P. fluorescens biovar 3 (6 strains), P. fluorescens biovar 4 (16 strains), Pseudomonas aureofaciens-Pseudomonas chlororaphis (3 strains), Pseudomonas aeruginosa (4 strains), Pseudomonas glathei (2 strains), and Pseudomonas mephitica (2 strains). The P. fragi complex was further divided into subclusters; the major subcluster (comprising 93 strains, including the type strain) was regarded as P. fragi sensu stricto. P. fluorescens and allied bacteria closely matched the descriptions given by Stanier et al. (J. Gen. Microbiol. 43:159-271, 1966). The characteristics for the 10 significant clusters are given. Also given are criteria which differentiate the P. fragi subclusters. The phylogenetic relationships among the meat-associated taxa were calculated. P. fluorescens biovars 2 and 3 were clearly separated from the remaining taxa. Biovar 4 is the most conservative, while biovar 3 has evolved at the highest rate.

Since type strain B348 of Eubacterium cellulosolvens (Cillobacterium cellulosolvens Bryant, Small, Bouma, and Robinson) was lost, we propose that van Gylswyk and Hoffman strain 6 (= ATCC 00000) be designated the neotype strain of this species. The characteristics of the neotype strain are compared with the characteristics of strains described elsewhere and with some of the most important characteristics of strains recently isolated by us from sheep rumina.

Vibrio strains isolated from plankton, sediments, and seawater in two coastal areas south of Valencia, Spain, were genotypically and phenotypically different from other Vibrio species. The guanine-plus-cytosyine content of the strains was 43 mol%. Deoxyribonucleic acid-deoxyribonucleic acid hybridization studies revealed that the strains were genetically homogeneous and exhibited very low levels of genetic relatedness to Vibrio parahaemolyticus ATCC 17802T(T = type strain) (0%), Vibrio harveyi NCMB 1280T(11%), Vibrio vulnificus NCMB 2046T(7%), Vibrio nereis NCMB 1897T(0%), Vibrio natriengens NCMB (9%), and Vibrio splendidus NCMB 1T(0%). Strain CECT 621 of the Colección Española de Cultivos Tipo was designated the type strain of the proposed new species Vibrio mediterranei.

We propose a new species, Alcaligenes piechaudii; the type strain is strain CIP 60.75 (= Hugh 366-5 = IAM 12591 = LMG 1873). Each of the seven strains placed in this new species was examined for 171 phenotypic characters, including reactions in 68 enzyme tests (API ZYM system). All of the strains displayed similar protein electrophoretic patterns and yielded low deoxyribonucleic acid relatedness values with the type strains of Alcaligenes faecalis, Alcaligenes denitrificans, and Achromobacter xylosoxidans. The results of deoxyribonucleic acid-ribosomal ribonucleic acid hybridization experiments localized Alcaligenes piechaudii on the Alcaligenes denitrificans ribosomal ribonucleic acid branch in the direct vicinity of Alcaligenes denitrificans and Achromobacter xylosoxidans. Alcaligenes piechaudii strains are rod shaped, aerobic, gram negative, nonpigmented, motile by peritrichous flagella, and nonsaccharolytic and reduce nitrate but not nitrite. The guanine-plus-cytosine contents of the deoxyribonucleic acids of the seven strains ranged from 64 to 65 mol%. The strains of the new species were isolated primarily from human clinical specimens, but some strains were also isolated from the environment. The validated but illegitimate name Alcaligenes denitrificans subsp. xylosoxydans (Yabuuchi and Yano 1981) Kersters and De Ley 1984 is corrected with a formal proposal of the proper combinations Alcaligenes xylosoxidans subsp. xylosoxidans (Yabuuchi and Yano 1981) comb. nov. (type strain, strain ATCC 27061) and Alcaligenes xylosoxidans subsp. denitrificans (Rüger and Tan 1983) comb. nov. (type strain, strain ATCC 15173).

The three type strains and a reference strain of Mobiluncus examined in this study had a gram-positive type of cell walls. All four strains were susceptible to vancomycin and resistant to colistin, and the overall susceptibility patterns with other antimicrobial agents were consistent with those of gram-positive microorganisms. Strains of Mobiluncus mulieris were lysed by 3% potassium hydroxide, and strains of Mobiluncus curtisii were not. However, all of the strains examined had intermediate levels of Limulus amebocyte-lysate reactivity that was not lipopolysaccharide associated. Lipopolysaccharide was not detected when whole-cell lysates were digested with proteinase K, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and silver stained. In addition, 2-keto-3-deoxyoctulosonic acid and heptose were not detected by gas chromatography of O-methyloxime acetate derivatives of whole-cell carbohydrates. Saponification and gas chromatography of whole-cell fatty acids showed that none of the four strains examined had detectable levels of hydroxylated fatty acids. Hydroxylated fatty acids or aldehyde fatty acids were not detected in acid hydrolysates of crude cell membrane preparations.

Methane-producing bacterial strain MS2T(T = type strain) was isolated from a tannery by-products enrichment culture inoculated with sewage sludge. This methanogen was a non-motile, irregular coccoid organism (diameter, 1 to 2 μm) which used H2-CO2 and formate as methanogenic substrates. Acetate was required for growth but did not serve as a methanogenic substrate; yeast extract and Trypticase peptone were highly stimulatory. Growth occurred through the pH range from 5.5 to 8.0, with optimum growth at pH 6.7. The optimum temperature for growth was 37°C. The deoxyribonucleic acid base composition was 59 mol% guanine plus cytosine. The physiological and antigenic properties of this isolate place it in the genus Methanogenium. The organism is named Methanogenium bourgense sp. nov.

A group of Beggiatoa-like organisms which do not deposit sulfur from sulfide but which were isolated from the same locations as sulfide-oxidizing beggiatoas should be considered as a species within the genus Vitreoscilla. These organisms fit the general description of Vitreoscilla beggiatoides as originally described by E. G. Pringsheim, and strain B23SS (= ATCC 43189) is herein designated as the neotype strain. The guanine-plus-cytosine content of the deoxyribonucleic acid of strain B23SS was 42 mol%. A few strains that were formerly considered as Beggiatoa spp. have been shown to lack the ability to accumulate sulfur in inclusions when exposed to hydrogen sulfide. Because rigorous tests were not previously carried out to determine whether these organisms really deposited sulfur, it is probable that they were never capable of doing so. These strains, which form a distinct group of microorganisms, now should be considered to be a Vitreoscilla species. This group of organisms is most characteristic of the species V. filiformis as originally described by E. G. Pringsheim. Because strains had not been characterized since the original description, V. filiformis was not included in the Approved List of Bacterial Names. This name has, therefore, been revived, and strain L1401-2 (= ATCC 43190), a strain originally isolated by E. G. Pringsheim, is designated as the neotype strain. The guanine-plus-cytosine content of the deoxyribonucleic acid of these strains ranged from 59 to 63 mol%. These two groups of organisms are compared to V. stercoraria, the best-characterized Vitreoscilla strain, and to Beggiatoa alba, the best-characterized and type species of the genus Beggiatoa.

Acetivibrio cellulosolvens BASTwas compared with Acetivibrio cellulolyticus CD2Tand was found to be identical in all morphological and biochemical characteristics. The description of the genus Acetivibrio is emended as originally described by I. M. Robinson and A. E. Ritchie (Int. J. Syst. Bacteriol. 31:333-338, 1981) to include anaerobic, gram-negative, straight to slightly curved rods that produce mainly acetic acid, ethanol, H2 and CO2 and that are motile by means of a single flagellum or multiple flagella. It is proposed that A. cellulosolvens BASTbe correctly classified as A. cellulolyticus.

A new species, Acetobacter methanolicus, is described. The strains investigated were isolated from sludge and from a yeast fermentation process in which methanol was the sole source of carbon and energy. A total of about 140 phenotypic features were tested. The strains proved to be acidophilic and facultatively methylotrophic, and they differed from other Acetobacter species by growing well on methanol, glucose, gluconate, 2,3-butanediol, and caproic acid as sole sources of carbon and energy. Ethanol was “overoxidized” only at initial concentrations of <0.5%. Lactate was oxidized very weakly, but it was not utilized as a sole carbon source for growth. Yeast extract or pantothenic acid was essential for growth. The specific epithet of the proposed new species refers to its isolation from media in which methanol was the sole source of carbon. The deoxyribonucleic acid base composition of type strain MB58 (= IMET 10945) is 62.3 mol% guanine plus cytosine.

Filter-fixed deoxyribonucleic acid preparations from organisms of the Neisseriaceae and from two unpigmented, psychrophilic, misnamed “Achromobacter” strains were hybridized with labeled ribosomal ribonucleic acids (rRNAs) from Acinetobacter calcoaceticus ATCC 23055T(T = type strain), Alteromonas macleodii ATCC 27126T, Escherichia coli B, Pasteurella multocida NCTC 10322T, Actinobacillus lignieresii NCTC 4189T, Vibrio parahaemolyticus ATCC 17802T, Pseudomonas fluorescens MMCA 40T, Deleya aquamarina NCMB 557, Marinomonas vaga ATCC 27119T, Xanthomonas campestris NCPPB 528T, Alcaligenes denitrificans ATCC 15173T, Pseudomonas solanacearum NCPPB 325T, Pseudomonas acidovorans ATCC 15668T, Alcaligenes faecalis NCIB 8156T, Janthinobacterium lividum NCTC 9796T, Chromobacterium violaceum NCTC 9757T, Acetobacter aceti NCIB 8621T, Agrobacterium tumefaciens ICPB TTIII, and Rhizobium meliloti NZP 4009. The results revealed that the present members of the Neisseriaceae are genetically very heterogeneous. This family contains at least five unrelated groups. One group is located in rRNA superfamily III (containing the Pseudomonas acidovorans complex, Alcaligenes, Janthinobacterium, etc.) and includes the “true neisseriae,” Kingella kingae, and Kingella denitrificans. The present emended family Neisseriaceae should be limited to these organisms. Its closest genetic relative is Chromobacterium. Another group, formed by Acinetobacter, Moraxella, Branhamella, the “false neisseriae,” and the misnamed achromobacters, should be removed from the Neisseriaceae; it is somewhat related to organisms belonging to rRNA superfamily II (containing the Pseudomonas fluorescens complex, Marinomonas, etc.). Kingella indologenes does not belong to either of these groups and is situated between rRNA superfamily I (containing the Enterobacteriaceae, Pasteurellaceae, etc.) and rRNA superfamily II. Moraxella urethralis and Moraxella anatipestifer are not at all related to other members of the genus Moraxella.

A new species of rapidly growing, nonphotochromogenic mycobacteria, Mycobacterium moriokaense, is described. This taxon commonly occurs in the soil and may have been erroneously identified previously as Mycobacterium fortuitum. M. moriokaense can be differentiated from all previously described species of mycobacteria. The type strain is strain ATCC 43059 (= NCH E11715).

We propose the name Pseudomonas lundensis for a new species of gram-negative, polarly flagellated, chemoorganotrophic, rod-shaped bacteria that were isolated from refrigerated meat. Strains of P. lundensis are capable of respiratory but not fermentative metabolism; they grow at 0°C, produce fluorescent pigments, catalase, and cytochrome c oxidase, and possess an arginine dihydrolase system. The mean guanine-plus-cytosine content of the deoxyribonucleic acids (DNAs) of 12 strains was 59.1 mol% (standard deviation, 0.7 mol%). Numerical phenotypic studies performed with 60 strains and DNA-DNA hybridization experiments with 12 strains (data separately published) revealed a tight phenotypic and genotypic cluster. P. lundensis is related to Pseudomonas fragi and the Pseudomonas fluorescens DNA-DNA homology group, and characteristics which differentiate these organisms and Pseudomonas aeruginosa are given. The type strain is strain 138 of Molin and Ternström (= 573 = CCM 3503); it was isolated from beef and has a guanine-plus-cytosine content of 60 mol %.

Because deoxyribonucleic acid-deoxyribonucleic acid hybridization has shown that the species Pasteurella ureae belongs to the genus Actinobacillus Brumpt 1910, we propose the transfer of P. ureae to the genus Actinobacillus under the new combination Actinobacillus ureae. The type strain of A. ureae is strain S. D. Henriksen 3520/59 (= ATCC 25976 = NCTC 10219).

The taxonomic relationships among Streptococcus anginosus type strain NCTC 10713, beta-hemolytic streptococci belonging to serological groups A, C, G, and F, and related organisms were determined by using deoxyribonucleic acid-deoxyribonucleic acid hybridization (S1 nuclease method), deoxyribonucleic acid base composition, and conventional biochemical tests. Minute-colony-forming beta-hemolytic streptococci which carry various Lancefield grouping antigens (antigens A, C, F, and G) or none of the known group antigens were closely related genetically and biochemically to the type strain of the S. anginosus, which carries group G antigen.

Campylobacter sputorum biovar sputorum ATCC 35980 is designated the neotype strain of the species.

Bacteroides termitidis (Sebald) differs so much from the type species of the genus Bacteroides, Bacteroides fragilis (Castellani and Chalmers), that it should not be retained within this genus. On the basis of biochemical, chemical, and genetic criteria, we propose that Bacteroides termitidis be reclassified in a new genus, Sebaldella, as Sebaldella termitidis comb. nov., the type species of the genus. The type strain of S. termitidis is strain NCTC 11300 (= ATCC 33386).

We propose that the photosynthetic procaryotes containing chlorophylls a and b in the species Prochloron didemni sp. nov., genus Prochloron gen. nov., be placed under the International Code of Nomenclature of Bacteria by including the genus Prochloron in the family Prochloraceae fam. nov., order Prochlorales ord. nov. in the class Photobacteria Gibbons and Murray 1978, listed on the approved lists of bacterial names.