- Volume 33, Issue 3, 1983

Strain CIP 82.01 of Vibrio alginolyticus and strain I.029 (previously designated “Achromobacter iophagus”) were compared. Strain I.029 produces a collagenase of high specific activity (Achromobacter collagenase; EC 3.4.24.8). Collagenase production is induced in strain CIP 82.01 by collagen or macromolecular fragments of collagen in a manner similar to collagenase induction in strain I.029; caseinolytic proteinase is constitutive. In this study we demonstrate that both strains also produce a constitutive extracellular endonuclease. Collagenases from both strains cleave either native collagen in its helical region or a similar synthetic peptide; both enzymes are inhibited by ethylenediamine tetraacetate, but not by diisopropyl fluorophosphate. The collagenase subunit (molecular weight, 35,000) of strain CIP 82.01 is similar in amino acid composition to the subunit of the strain I.029 enzyme, although some of the aspartic and threonine residues in strain CIP 82.01 are replaced by glutamic and serine residues in strain I.029. Surface radioiodination followed by two-dimensional electrophoresis showed that there are quantitative differences in the major outer membrane proteins of the two strains. Strains CIP 82.01 and I.029 differ qualitatively in resistance to ampicillin and carbenicillin, in cellobiose fermentation, in ornithine decarboxylase activity, and in halophilism. We propose that strain I.029, which was originally designated A. iophagus, be included within the species V. alginolyticus, but that this organism be distinguished from other strains of this species by the designation Vibrio alginolyticus chemovar iophagus, with the corresponding collagenase designated “iophagus collagenase” (EC 3.4.24.8).

A numerical analysis of phenetic data collected from rapidly growing, scotoch-romogenic mycobacterial strains isolated from water in South Africa and from strains of the taxa Mycobacterium parafortuitum, Mycobacterium aurum, Mycobacterium neoaurum, and “Mycobacterium diernhoferi” indicated that all of these organisms belong to the Mycobacterium parafortuitum complex. Our results also indicated that each of the taxa mentioned above is worthy of species status within the complex. The name Mycobacterium austroafricanum sp. nov. is proposed for the South African isolates, and the characteristics of these isolates are described; the type strain is E9789-SA12441 (= ATCC 33464). The name Mycobacterium diernhoferi is revived for the organism described originally by Bönicke and Juhasz in 1965; the type strain of this species is strain 41001 (= ATCC 19340).

We examined 47 Hafnia alvei strains by performing a numerical analysis of 101 features. The similarity among all of the strains was very high, and these strains formed a single phenon with a simple matching coefficient of more than 0.875 when average-linkage cluster analysis was used. The H. alvei strains were distinct from several reference enteric bacteria, including representative Enterobacter strains. The guanosine-plus-cytosine contents of the deoxyribonucleic acids from nine strains were 43.6 to 47.9 mol% (mean ± standard deviation, 45.3 ± 1.3 mol%).

Five rapidly growing, glucose-fermenting Mycoplasma strains were isolated from the lungs of mink kits. Biochemical and serological studies revealed that these isolates were different from all of the currently accepted Mycoplasma species and serogroups. These strains appear to constitute a new species, for which we propose the name Mycoplasma mustelae. The type strain of M. mustelae is strain MX9, and this strain has been deposited in the Food and Agriculture Organization/World Health Organization Collaborating Centre for Animal Mycoplasmas, Institute of Medical Microbiology, University of Aarhus, Aarhus, Denmark, as strain AMRC-C 1486, in the American Type Culture Collection, Rockville, Md., as strain ATCC 35214, and in the National Collection of Type Cultures, London, England, as strain NCTC 10193.

We describe a new species, Staphylococcus gallinarum, based on a study of characteristics of strains isolated from chickens and a pheasant. Strain CCM 3572 (= VIII1) is the type strain of this species. The chemical composition of the cell walls of the strains of this new species is similar to that of Staphylococcus epidermidis, but these organisms are resistant to novobiocin and produce acid from a wide range of carbohydrates. Staphylococcal strains isolated from goat milk belong to another new species, Staphylococcus caprae. The type strain is CCM 3573 (= 143.22). These strains are more closely related to S. epidermidis and Staphylococcus capitis than to other staphylococci.

We hybridized 23S 2-14C-labeled ribosomal ribonucleic acids (rRNAs) from type strains Pseudomonas fluorescens ATCC 13525, Pseudomonas acidovorans ATCC 15668, Pseudomonas solanacearum NCPPB 325, and Xanthomonas campestris NCPPB 528 with deoxyribonucleic acids (DNAs) from 65 Pseudomonas strains, 23 Xanthomonas strains, and 148 mostly gram-negative strains belonging to 43 genera and 93 species and subspecies including more than 60 type strains. Our findings confirm and extend the findings derived from ribonucleic acid hybridizations by the Berkeley group, but differed in some respects from the groupings of Pseudomonas in Bergey’s Manual of Determinative Bacteriology, 8th ed. The genus Pseudomonas Migula 1894, 237 was divided into three large, distinct groups. The Pseudomonas fluorescens rRNA branch contains Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas chlororaphis, Pseudomonas aureofaciens, Pseudomonas syringae, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas mendocina, Pseudomonas cichorii, Pseudomonas alcaligenes, and Pseudomonas pseudoalcaligenes. The Pseudomonas acidovorans rRNA branch contains Pseudomonas acidovorans, Pseudomonas testosteroni, Pseudomonas delafieldii, Pseudomonas facilis, Pseudomonas palleronii, Pseudomonas saccharophila, and Pseudomonas flava. The third rRNA branch contains Pseudomonas solanacearum, Pseudomonas cepacia, Pseudomonas marginata, Pseudomonas caryophylli, and Pseudomonas lemoignei. Each of these rRNA branches is as heterogeneous as a genus. The Pseudomonas solanacearum and Pseudomonas acidovorans rRNA branches are about as far removed from each other as they are from the genera Janthinobacterium and Derxia and the authentic genus Alcaligenes. These branches are members of the third rRNA superfamily. The Pseudomonas fluorescens rRNA branch is quite different, as it is a member of the second rRNA superfamily, which also contains Azotobacter, Azomonas, Xanthomonas, and some other genera. Along with data from rRNA hybridizations involving many different gram-negative taxa, these results show clearly that the three Pseudomonas rRNA branches differ at least at the genus level. The genus Xanthomonas is separate in its own right. It constitutes a very tight cluster consisting of Xanthomonas campestris, Xanthomonas fragariae, Xanthomonas axonopodis, and Xanthomonas albilineans (Xanthomonas campestris covers older species names no longer in use). Xanthomonas (Aplanobacter) populi has rRNA cistrons that are indistinguishable from the rRNA cistrons of the xanthomonads mentioned above. There are a number of misnamed taxa. Pseudomonas maltophilia is a somewhat unusual member of Xanthomonas; likewise, Pseudomonas diminuta and Pseudomonas vesicularis are not members of the genus Pseudomonas, and Xanthomonas ampelina is definitely not a member of the genus Xanthomonas. The exact taxonomic positions of the latter three species are unknown. A quantitative comparison showed that fine differentiation of strains by means of DNA-DNA hybridization under stringent conditions at Tor (temperature of optimal renaturation) was meaningful only in the top 7 to 8°C T m(e) (thermal elution temperature range, 73 to 81°C) of our DNA-rRNA similarity maps and dendrograms (a difference of 1°C in thermal elution temperature T m(e) from ribosomal DNA similarity corresponded to roughly 14% DNA homology).

The Pasteurella-like organism of porcine necrotizing pleuropneumonia described by Bertschinger and Seifert in 1978 was shown to be phenotypically similar to the V factor-requiring strains hitherto classified as Haemophilus pleuropneumoniae (Matthews and Pattison 1961) Shope 1964. Deoxyribonucleic acid-deoxyribonucleic acid hybridization experiments in which the optical method was used yielded 91% binding of the genomes of the type and reference strains of these two taxa, thus indicating that they belong to the same species. The levels of deoxyribonucleic acid binding between these strains and the type strain of Haemophilus influenzae indicated no measurable relatedness. Therefore, these organisms do not belong to the genus Haemophilus sensu stricto. On the other hand, they are closely related to Actinobacillus lignieresii, as determined by both phenotypic characteristics and deoxyribonucleic acid base sequence relatedness (72 to 75% binding). To take into account the different host ranges of these organisms, we propose to classify them as a species distinct from, but closely related to, A. lignieresii, under the new combination Actinobacillus pleuropneumoniae. The emended species A. pleuropneumoniae is composed of V factor-requiring and V factor-independent biovars. The type strain (strain Shope 4047 [= CCM 5869 = ATCC 27088]) requires V factor; the V factor-independent biovar is represented by strain Bertschinger 2008/76 (= Frederiksen P 597 = HIM 677-3/4).

Three gram-negative, oxidase-positive diplococcal strains were isolated from the oropharynges of healthy monkeys. These three strains closely resembled Neisseria perflava in their physiological and biochemical characteristics, were more similar to Neisseria canis in their cellular fatty acid profiles, and were moderately related to Neisseria mucosa (51.9%) as determined by deoxyribonucleic acid-deoxyribonucleic acid hybridization. An analysis of 11 enzymes indicated clustering closest to Neisseria sicca, followed by N. mucosa. We propose the name Neisseria macacae for this new species, and the type strain of this species is strain M-740 (= ATCC 33926).

We assessed polynucleotide sequence homologies among representative strains from four genera of ammonia-oxidizing bacteria, including three morphological types of the genus Nitrosomonas, by deoxyribonucleic acid-deoxyribonucleic acid hybridization. Our results indicate that there is little homology among the four genera which we examined. Furthermore, the low degree of homology among the morphological types of Nitrosomonas suggests that each type should be considered a separate species.

Promicromonospora enterophila sp. nov., which was isolated from millipede feces, is the first species of the genus Promicromonospora for which the natural habitat is known. A comparison of this species with Promicromonospora citrea is presented. The type strain of P. enterophila is HMGB B1078 (= strain DFA-19).

Thiobacillus perometabolis THI 024 (= ATCC 27793) is closely related to Thiobacillus sp. strain A2T (type strain) (= THI 041T = ATCC 25364T), and these strains clearly differ from the known Thiobacillus species in their morphological and physiological characteristics, ubiquinone systems, deoxyribonucleic acid guanine-plus-cytosine contents, and cellular fatty acid compositions. We propose the name Thiobacillus rapidicrescens sp. nov. for these organisms. Strain THI 041 is the type strain of this species.

A bacterial disease characterized by the development of rectangular lesions on the leaves of konjac (Amorphophalus konjac Koch.) is described. The size of the lesions increased rapidly under humid conditions, resulting in leaf blight. Leaves with severe blight drooped and finally decayed. The disease was identified from the symptoms as bacterial leaf blight on the basis of the descriptions made by Uyeda in 1910. The causal agent was, however, different from Xanthomonas conjaci (Uyeda) Burkholder in many respects and was found to be a new subspecies of Pseudomonas pseudoalcaligenes based on the results of a direct comparison of pathological and biochemical properties. The name Pseudomonas pseudoalcaligenes subsp. konjaci subsp. nov. is proposed for the new subspecies, and strain K2 (= ATCC 33996 = PDDCC 7733) is designated the type strain.

Pseudomonas ficuserectae, a new nonfluorescent, phytopathogenic pseudomonad species, is described. This bacterium produces dark brown, water-soaked spots on the leaves and stems of Ficus erecta Thunb., resulting either in defoliation or shoot blight on severely infected plants. The colonies on nutrient agar plates are white, circular, and 3 to 4 mm in diameter after 6 days. P. ficuserectae is similar to Pseudomonas amygdali in many properties. The differences between these two species include size of colonies on agar plates, hydrolysis of Tween 80, production of H2S, utilization of ribose, raffinose, mannitol, sorbitol, and malonate, and pathogenicity. The deoxyribonucleic acid base composition of the type strain of P. ficuserectae, strain L7, is 59 mol% guanine plus cytosine. The specific epithet of this new species reflects the pathogenicity of the bacterium on F. erecta Thunb.

Strains R1T (type strain) and R2, two new isolates of nonsulfur purple photosynthetic bacteria, were studied. The cells of these organisms were gram-negative nonsporeforming rods that were motile by means of a single flagellum, and they multiplied by budding. The photosynthetic membrane systems of these strains were of the lamellar type parallel to the cytoplasmic membrane. Cells which formed under anaerobic conditions in the light were red. The cells of these strains contained bacteriochlorophyll a, ubiquinone 10, and carotenoids. No vitamins were required as growth factors, and reduced sulfur compounds could not be utilized by strains R1T and R2. These two strains produced molecular hydrogen anaerobically in the light. On the basis of this study, we propose a new species, Rhodopseudomonas rutila, for strains R1T and R2. The guanine-plus-cytosine contents of the deoxyribonucleic acids of strains R1T and R2 are 67.6 and 69.4 mol%, respectively. The type strain of this new species is strain R1 (= ATCC 33872).

A new species of Actinosporangium is described, for which we propose the name Actinosporangium vitaminophilum. This organism produces new antibiotics called pyrrolomycins and is characterized by pseudosporangia lacking sporangial walls, nonmotile spores, and a type I cell wall. It differs significantly from previously described species of the genus Actinosporangium in the cultural conditions necessary for pseudosporangium formation. The type strain of A. vitaminophilum is SF-2080 (= ATCC 31673).

We tested the possible pathogenicity of 27 bacteria that are budding or prosthecate or both in mice and guinea pigs. The strains tested belong to the following taxa: Hyphomicrobium (12 strains), “Hyphomonas” (4 strains), Pedomicrobium (2 strains), Rhodomicrobium (2 strains), Prosthecomicrobium (1 strain), “Stella” (1 strain), Blastobacter (2 strains), Planctomyces (1 strain), and Pasteuria (1 strain); 1 strain was unnamed. Mice injected intraveneously with a 0.5-ml suspension containing up to 109 colony-forming units per ml survived for 7 days without macroscopic changes, abscesses, or lesions. Four Hyphomicrobium strains, one “Hyphomonas” strain, and the unnamed strain caused slight spleen enlargement and in some cases a mottled kidney appearance; however, the organs were histologically normal, and the changes observed were considered to be signs of host reaction rather than pathological changes. Subcutaneous injections also did not result in disease. Two Rhodomicrobium strains could be reisolated from mice after 21 days; none of the other strains could be recovered after 7 days. Lethal endotoxic activity was not detected with any of the strains when cell sonic extracts were injected intraveneously. All strains caused antibody formation, and agglutination dilution titers ranged from 1 :2 to 1 :64. Serological cross-reactions with six strains indicated the presence of distinct taxonomic groups. Hyphomicrobia from soil cross-reacted with each other but not with hyphomicrobia from aquatic sources. Immunosuppressive pretreatment with Natulan did not render the mice more susceptible. Other possible indicators of pathogenicity (i.e., mannitol utilization and β-hemolysis) were not observed in the 27 strains. We concluded that the 27 bacteria tested are nonpathogenic for mice or guinea pigs under the conditions used.

Four strains of obligately anaerobic spirochetes were isolated from marine intertidal mud. These strains grew to yields of 2 × 108 to 6 × 108 cells per ml in chemically defined media containing glucose, amino acids or NH4Cl, sulfide, vitamins, and inorganic salts. These spirochetes required relatively high levels of Na+ for growth and utilized carbohydrates but not amino acids as growth substrates. Glucose was fermented to CO2, H2, ethanol, and acetate. These four strains also formed the branched-chain fatty acids isovalerate , 2-methylbutyrate, and isobutyrate as fermentation products of l-leucine, l-isoleucine, and l-valine, respectively. The adenosine 5'-triphosphate generated via fermentation of these amino acids was expended by the spirochetes to prolong survival during periods of growth substrate starvation. The deoxyribonucleic acids of the four strains contained 63.6 to 65.6 mol% guanine plus cytosine. We concluded that these four strains are representatives of a new species, for which we propose the name Spirochaeta isovalerica; the type strain of this species is strain MA-2 (= DSM 2461).

A new genus, Sphingobacterium, two new combinations, Sphingobacterium spiritivorum (Holmes, Owen, and Hollis) and Sphingobacterium multivorum (Holmes, Owen, and Weaver), and two new species, Sphingobacterium mizutae and Flavobacterium indologenes, are described. The genus Sphingobacterium is characterized and distinguished from the genus Flavobacterium by the presence in its strains of high concentrations of sphingophospholipids as cellular lipid components. S. spiritivorum is designated the type species of the genus Sphingobacterium. S. mizutae ATCC 33299 (= GIFU 1203) and F. indologenes ATCC 29893 (= CDC 3716 = GIFU 1347 = NCTC 10796 = RH 542) are designated the type strains of the two new species. Emended descriptions of the type strains of S. spiritivorum and S. multivorum are presented.

During a survey for the presence of Aeromonas salmonicida, 45 Aeromonas isolates which could not be classified in the existing named species of the genus were recovered from water obtained from the River Avon in Hampshire, England. On the basis of a numerical taxonomy study, these isolates were clustered in three homogeneous phena. Phenon 1 was identified as a new species of Aeromonas, for which we propose the name Aeromonas media; strain RM (= ATCC 33907) is the type strain of this species.