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Taxonomy of aerobic marine eubacteria.

Two hundred and eighteen strains of nonfermentative marine bacteria were submitted to an extensive morphological, physiological, and nutritional characterization. All the strains were gram-negative, straight or curved rods which were motile by means of polar or peritrichous flagella. A wide variety of organic substrates served as sole sources of carbon and energy. The strains differed extensively in their nutritional versatility, being able to utilize from 11 to 85 carbon compounds. Some strains had an extracellular amylase, gelatinase, lipase, or chitinase and were able to utilize n-hexadecane and to denitrify. None of the strains had a yellow, cell-associated pigment or a constitutive arginine dihydrolase system, nor were they able to hydrolyze cellulose or agar. The results of the physiological and nutritional characterization were submitted to a numerical analysis which clustered the strains into 22 groups on the basis of phenotypic similarities. The majority of these groups were separable by a large number of unrelated phenotypic traits. Analysis of the moles per cent guanine plus cytosine (GC) content in the deoxyribonucleic acid of representative strains indicated that the peritrichously flagellated groups had a GC content of 53.7 to 67.8 moles%; polarly flagellated strains had a GC content of 30.5 to 64.7 moles%. The peritrichously flagellated groups were assigned to the genus Alcaligenes. The polarly flagellated groups, which had a GC content of 43.2 to 48.0 moles%, were placed into a newly created genus, Alteromonas; groups which had a GC content of 57.8 to 64.7 moles% were placed into the genus Pseudomonas; and the remaining groups were left unassigned. Twelve groups were given the following designations: Alteromonas communis, A. vaga, A. macleodii, A. marinopraesens, Pseudomonas doudoroffi, P. marina, P. nautica, Alcaligenes pacificus, A. cupidus, A. venustus, and A. aestus. The problems of assigning species of aerobic marine bacteria to genera are discussed.

Alcaligenes↗

Diverse enzymological patterns of phenylalanine biosynthesis in pseudomonads are conserved in parallel with deoxyribonucleic acid homology groupings.

l-Tyrosine biosynthesis in nature has proven to be an exceedingly diverse gestalt of variable biochemical routing, cofactor specificity of pathway dehydrogenases, and regulation. A detailed analysis of this enzymological patterning of l-tyrosine biosynthesis formed a basis for the clean separation of five taxa among species currently named Pseudomonas, Xanthomonas, or Alcaligenes (Byng et al., J. Bacteriol. 144:247-257, 1980). These groupings paralleled taxa established independently by ribosomal ribonucleic acid/deoxyribonucleic acid (DNA) homology relationships. It was later found that the distinctive allosteric control of 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase in group V, a group dominated by most named species of Xanthomonas (Whitaker et al., J. Bacteriol. 145:752-759, 1981), was the most striking and convenient criterion of group V identity. Diversity in the biochemical routing of l-phenylalanine biosynthesis and regulation was also found, and phenylalanine patterning is in fact the best single enzymatic indicator of group IV (Pseudomonas diminuta and Pseudomonas vesicularis) identity. Enzymological patterning of l-phenylalanine biosynthesis allowed discrimination of still finer groupings consistently paralleling that achieved by the criterion of DNA/DNA hybridization. Accordingly, the five ribosomal ribonucleic acid/DNA homology groups further separate into eight DNA homology subgroups and into nine subgroups based upon phenylalanine pathway enzyme profiling. (Although both fluorescent and nonfluorescent species of group I pseudomonads fall into a common DNA homology group, fluorescent species were distinct from nonfluorescent species in our analysis.) Hence, phenylalanine patterning data provide a relatively fine-tuned probe of hierarchical level. The combined application of these various enzymological characterizations, feasibly carried out in crude extracts, offers a comprehensive and reliable definition of 11 pseudomonad subgroups, 2 of them being represented by species of Alcaligenes.

Alcaligenes↗

Plasmid-mediated mineralization of 4-chlorobiphenyl.

Strains of Alcaligenes and Acinetobacter spp. were isolated from a mixed culture already proven to be proficient at complete mineralization of monohalogenated biphenyls. These strains were shown to harbor a 35 X 10(6)-dalton plasmid mediating a complete pathway for 4-chlorobiphenyl (4CB) oxidation. Subsequent plasmid curing of these bacteria resulted in the abolishment of the 4CB mineralization phenotype and loss of even early 4CB metabolism by Acinetobacter spp. Reestablishment of the Alcaligenes plasmid, denoted pSS50, in the cured Acinetobacter spp. via filter surface mating resulted in the restoration of 4CB mineralization abilities. 4CB mineralization, however, proved to be an unstable characteristic in some subcultured strains. Such loss was not found to coincide with any detectable alteration in plasmid size. Cultures capable of complete mineralization, as well as those limited to partial metabolism of 4CB, produced 4-chlorobenzoate as a metabolite. Demonstration of mineralization of a purified 14C-labeled chlorobenzoate showed it to be a true intermediate in 4CB mineralization. Unlike the mineralization capability, the ability to produce a metabolite has proven to be stable on subculture. These results indicate the occurrence of a novel plasmid, or evolved catabolic plasmid, that mediates the complete mineralization of 4CB.

Acinetobacter↗

Cellular fatty acid composition of group IVe, a nonsaccharolytic organism from clinical sources.

The cellular fatty acid composition of a group of gram-negative nonfermentative organisms designated group IVe was studied by gas-liquid chromatography. Strains of this group are isolated most frequently from urine and most closely resemble the Alcaligenes in conventional biochemical tests. On the basis of cellular fatty acids, however, we found these organisms to be strikingly different from Alcaligenes and other gram-negative species with similar phenotypic characteristics. The gas-liquid chromatography procedure offers an additional diagnostic test for rapid identification of unclassified bacteria like group IVe.

Alcaligenes↗

[Isolation of bacteria which could perform nitrification and denitrification simultaneously by gene probe].

The bacteria which could perform nitrification and denitrification simultaneously from nitrogen containing wastes in Taiwan were isolated by using the probes made from random DNA fragments of Thiosphaera pantotropha. Two isolates were identified and named Alcaligenes faecalis subsp. faecalis strain 1 and strain 2 respectively. The effects on nitrification and denitrification by different medium pH, oxygen content, addition of different electron donors or inhibitors were studied. The isolates not only could perform nitrification, but also denitrification even in the presence of oxygen. Potassium cyanide could inhibit denitrification; hydrazine and hydroxyamine could inhibit nitrification. Alcaligenes faecalis subsp. faecalis strain 2 shows better denitrification.

Alcaligenes↗

Conversion of solvent evaporation residues from the AB- (acetone - butanol) bioprocess into bacterial cells accumulating thermoplastic polyesters.

In a bioconversion study based on utilisation of by-products from the AB- (acetone - butanol) bioprocess a new isolated gram-negative solvent tolerant bacterium was used to convert the AB process residue after removal of the major part of the solvents. The bacterium identified as a representative of the genus Alcaligenes (designated as Alcaligenes sp. G) was capable of growth up to optical densities ranging from 8 to 20 and simultaneously of polyhydroxyalkanoate-(PHA-)accumulation up to 40% per dry weight. A standardised medium based on AB by-products containing 7 g/l of butyrate and 5 g/l of acetate at pH 7.5 was used in our studies for bioconversion into PHAs. Concentrations of 1-butanol, which is known for its membrane damaging properties in microorganisms, were tolerated in the AB by-products medium up to 4 g/l without significant inhibition of cellular growth. No inhibition of growth was observed, when the medium was adjusted to 40 g/l butyrate. Due to the toxicity of the remaining 1-butanol maintenance of sterility is of no high priority during the process. The use of acetate and butyrate from an AB process is expected to provide a higher return-on-investment than the combustion of biogas to help meet energy demands.

Acetone↗

Conversion of solvent evaporation residues from the AB- (acetone-butanol) bioprocess into bacterial cells accumulating thermoplastic polyesters.

In a bioconversion study based on utilisation of by-products from the AB- (acetone-butanol) bioprocess a new isolated gram-negative solvent tolerant bacterium was used to convert the AB process residue after removal of the major part of the solvents. The bacterium identified as a representative of the genus Alcaligenes (designated as Alcaligenes sp. G) was capable of growth up to optical densities ranging from 8 to 20 and simultaneously of polyhydroxyalkanoate- (PHA-)accumulation up to 40% per dry weight. A standardised medium based on AB by-products containing 7 g/l of butyrate and 5 g/l of acetate at pH 7.5 was used in our studies for bioconversion into PHAs. Concentrations of 1-butanol, which is known for its membrane damaging properties in micro-organisms, were tolerated in the AB by-products medium up to 4 g/l without significant inhibition of cellular growth. No inhibition of growth was observed, when the medium was adjusted to 40 g/l butyrate. Due to the toxicity of the remaining 1-butanol maintenance of sterility is of no high priority during the process. The use of acetate and butyrate from an AB process is expected to provide a higher return-on-investment than the combustion of biogas to help meet energy demands.

1-Butanol↗

[On the taxonomy and physiology of bacteria utilizing hydrocarbons in the sea (author's transl)].

Using mineral media with gas oil as sole carbon source, 191 bacterial strains were isolated from the costal area of Marseille. These strains were attributed to Achromobacter, Alcaligenes, Pseudomonas, Acinetobacter and Arthrobacter genera. Amongst isolated strains there was a predominance of the Alcaligenes-Achromobacter group over others genera. Growth and respiratory activity of 5 strains were studied on hexadecane and acetate. Respiratory activity on hexadecane of 32 strains cultured on acetate has been measured. These strains could be placed in two groups. The group 1 shows an immediate respiratory activity which is not abolished by chloramphenicol. The group 2 presents either an immediate or a delayed respiratory activity which is always abolished by chloramphenicol. Maintenance or suppression of respiration by chloramphenicol is a character which is homogenously distributed amongst the species.

Acetates↗

Compositional studies on succinoglycan-like extracellular water-soluble Rhizobium polysaccharides.

This study reports structural information on extracellular, water-soluble polysaccharides from 5 different strains of Rhizobium, viz. R. trifolii J60, R. meliloti J1017, 202, 204 and 207. All the 5 polysaccharides had glucose and galactose in approximate molar ratio of 7:1. Methylation analysis revealed that the polysaccharides contained (1 leads to 3), (1 leads to 6), (1 leads to 4), (1 leads to 4, 1 leads to 6)-linked D-glucose residues, (1 leads to 3)-linked D-galactose and non-reducing terminal D-glucose attached to pyruvate. This structure was found to be exactly the same as that of succinoglycan, a succinic acid containing water-soluble polysaccharide elaborated by Alcaligenes faecalis var. myxogenes 10C3. The similarity of the structure of polysaccharides of two different Rhizobium species and also to the polysaccharide produced by Alcaligenes are discussed in terms of host specificity.

Alcaligenes↗

Differentiation among bacteria isolated from turkeys with coryza (rhinotracheitis).

Gram-negative bacteria isolated from turkeys with coryza in the United States, the Federal Republic of Germany, and the Republic of South Africa were compared with known Alcaligenes species and Bordetella bronchiseptica. The turkey isolates were separated into three distinct groups based on biochemical and physiologic tests. Forty of the 68 isolates studied (group I) were different from Alcaligenes sp. and B. bronchiseptica. Isolates in group I produced a heat-labile hemagglutinin and did not grow on Simmons' citrate agar. Isolates in group II (25 isolates) were similar to A. faecalis and A. odorans, grew on Simmons' citrate agar, and did not produce a hemagglutinin. Isolates in group III were B. bronchiseptica. Isolates from groups I and II caused coryza in poults. Group III isolates were not pathogenic.

Alcaligenes↗

[Non-fermentative gram-negative bacilli: their distribution to clinical materials and antibiotic susceptibility (author's transl)].

A total of 7898 non-fermentative Gram-negative bacilli were isolated from various clinical materials. Pseudomonas (7526) was the most common among them. Alcaligenes faecalis (273), Acinetobacter sp. (93) and Flavobacterium (6) were the other non-fermentative Gram-negative bacilli. Most of them were found in urine and pus cultures, however they were also isolated from sputum, blood, and cerebrospinal fluid. Gentamicin was the most effective antibiotic in vitro. Fifty four per cent of Pseudomonas, 100% of Acinetobacter, and 70% of Alcaligenes faecalis were inhibited by tobramycin.

Acinetobacter↗

[Growth and substrate utilization by bacterial lawn on the agar surface: experiment and one-dimensional distributed model].

Cell mass dynamics of the lawns formed by Pseudomonas fluorescens and Alcaligenes sp. and the distribution profiles of the residual substrate in the agar layer were monitored. After one or two days of culturing, the concentration of pyruvate in the top agar layer adjacent to the lawn dropped below the level of detection, and, from this moment, the substrate was supplied to the lawn by diffusion from underlying agar layers. Diffusion of pyruvate in noninoculated bilayered agar was found to follow Fick's equation with the diffusion coefficient of 0.042 cm2/h. A distributed mathematical model adequately describing the growth of bacterial lawn was developed based on the diffusion equation and the Monod-Herbert kinetic model. Notable distinctions between the two cultures studied were revealed: pseudomonads had higher growth and death rates than Alcaligenes sp. and exhibited a greater affinity for the substrate.

Agar↗

Numerical taxonomy of Pseudomonas based on published records of substrate utilization.

Data published by R. Y. Stanier, N. J. Palleroni, M. Doudoroff and their colleagues on Pseudomonas have been analysed by numerical taxonomy. Records on 401 strains were used, representing 155 characters, mostly utilization of substrates as carbon-energy sources. Twenty-nine phenons were recognized, which included 394 strains: the remaining 7 remained unclustered. The results were in very good accord with the conclusions of these authors. Almost all phenons were well separated with very little overlap. Many of them corresponded to distinct species, and others corresponded to recognized biotypes. Some small groups may represent unnamed new species. Analyses by Gower's Coefficient showed five major groupings: A) the fluorescent pseudomonads; B) biochemically active species (Pseudomonas cepacia, P. pseudomallei and allies); C) moderately active free-living species (P. acidovorans, P. alcaligenes and allies); D) P. solanacearum and allies; and #) P. mallei P. diminuta does not appear to be clearly distinct from P. vesicularis, nor does P alcaligenes appear clearly distinct from P. pseudoalcaligenes. There may, however, be some difference between P. multivorans and P. cepacia. Analyses using the Pattern Coefficient differed mainly in the relationships shown by a few of the metabolically active species. Of the two coefficients, the Pattern Coefficient gave results that were in somewhat better agreement with evidence from nucleic acids, but it showed an unexpectedly close relationship between P. solanacearum and P. cepacia.

Fluorescence↗

Cloning and sequence analysis of the poly (3-hydroxyalkanoic acid)-synthesis genes of Pseudomonas acidophila.

Pseudomonas acidophila can grow with CO2 as a sole carbon source by the possession of a recombinant plasmid that clones genes that confer chemolithoautotrophic growth ability derived from the H2-oxidizing bacterium Alcaligenes hydrogenophilus. H2-oxidizing bacteria produce poly(3-hydroxybutyric acid) (PHB) from CO2, but recombinant P. acidophila can produce the more useful biopolymer poly(3-hydroxyalkanoic acid) (PHA). In this study, the pha genes of P. acidophila were cloned and a sequence analysis was carried out. A gene library was constructed using the cosmid vector pVK102. A recombinant cosmid carrying the pha genes was selected by the complementation of a PHB-negative mutant of Alcaligenes eutrophus H16. The resulting recombinant cosmid pIK7 contained a 14.8-kb DNA insert. Subcloning was done. and the recombinant plasmid pEH74 was selected by hybridization with the A. eutrophus H16 pha genes. Escherichia coli possessing pEH74 produced PHB, indicating that pEH74 contained the pha genes of P. acidophila. The nucleotide sequences of the PHA-synthesis genes phaA (beta-ketothiolase), phaB (acetoacetyl-CoA reductase), and phaC (PHA synthase) in pEH74 were determined. The homologies of phaA, phaB, and phaC between P. acidophila and A. eutrophus H16 were 64.7, 76.1 and 56.6%, respectively.

Acetyl-CoA C-Acyltransferase↗

[Correlation of 55 samples of contact lens fluid and direct swabbings of the eye surface of symptomatic wearers of contact lenses].

BACKGROUND: The microbiological examination of contact lens storage solutions may reveal the etiological organisms responsible for keratitis and additionally harbour environmental contaminants. PATIENTS AND METHODS: From 1995 to 1998, conjunctival and corneal smears and scrapes, as well as the storage solutions of 55 soft contact lens wearers were examined microbiologically in a prospective non-randomised cohort study involving 53x unilateral and 2x bilateral cases of keraitis. RESULTS: In 42 of the in-use contact lens storage solutions, the following organisms were found: 8x Bacillus sp., 7x P. aeruginosa and 8x other pseudomonads, 6x Alcaligenes xylosans, 4x Serratia liquefaciens, 4x Enterobacter sp., 4x coagulase negative staphylococci, 3x Serratia marcescens, 3x S. aureus, 2x Streptococcus faecalis, 1x Klebsiella oxytoca and 5x Acanthameba. A significant correlation of ocular cultures was found for Pseudomonas, Enterobacter, Serratia, S. aureus, S. faecalis and Acanthameba, but not for Bacillus, Alcaligenes, and coagulase negative staphylococci. Fungi were not found. The more pronounced the clinical picture, the more Gram-negative bacteria were isolated. CONCLUSIONS: The microbiological examination of soft contact lens storage solutions of 55 keratitis patients confirmed that the microbes responsible were to be found in the fluid, in addition to environmental contaminants.

Acanthamoeba Keratitis↗

Microbiology of high-sodium-nitrite-wastewater treatment.

A microbiological study conducted as a complement to kinetic studies of biological denitrification as a process for treating high-sodium-nitrite wastewaters generated from ship-boiler-tube cleaning is described. The number, genera, and denitrifying capabilities of the organisms inhabiting anoxic suspended-growth reactors used in the kinetic studies were evaluated for four experimental phases. The results regarding the enumeration of bacteria supported the findings of the kinetic studies as follows: (i) the better nitrite-removal efficiencies observed in the nitrification/denitrification system as compared with direct denitrification were confirmed by the presence of larger populations of organisms capable of completely reducing nitrate or nitrite; (ii) the presence of metals in concentrations associated with boiler-tube wastewater did not affect removal performance in the nitrification/denitrification systems, nor did it affect the density of complete denitrifiers; (iii) increasing sludge ages resulted in increasing nitrite-removal efficiencies as well as populations of complete denitrifiers; and (iv) a decrease in nitrate-removal efficiencies when the actual wastewater was introduced to a system that had been acclimated to the synthetic wastewater coincided with a reduction in the number of complete denitrifiers. Regarding the types of organisms found in this study, denitrifying strains of Alcaligenes and Pseudomonas were always present in the anoxic reactors along with other denitrifying and non-denitrifying bacteria of the same genera, or other genera such as Acinetobacter and Flavobacterium. However, members of the genus Alcaligenes were the only complete denitrifiers found in the anoxic reactors, and hence they are likely to play a key role in the denitrification process.

Journal Article↗

Bacterial communities and enzyme activities of PAHs polluted soils.

Three soils (i.e. a Belgian soil, B-BT, a German soil, G, and an Italian agricultural soil, I-BT) with different properties and hydrocarbon-pollution history with regard to their potential to degrade phenanthrene were investigated. A chemical and microbiological evaluation of soils was done using measurements of routine chemical properties, bacterial counts and several enzyme activities. The three soils showed different levels of polycyclic aromatic hydrocarbons (PAHs), being their contamination strictly associated to their pollution history. High values of enzyme activities and culturable heterotrophic bacteria were detected in the soil with no or negligible presence of organic pollutants. Genetic diversity of soil samples and enrichment cultures was measured as bands on denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA sequences from the soil and enrichment community DNAs. When analysed by Shannon index (H'), the highest genetic biodiversity (H'=2.87) was found in the Belgian soil B-BT with a medium-term exposition to PAHs and the poorest biodiversity (H'=0.85) in the German soil with a long-term exposition to alkanes and PAHs and where absence, or lower levels of enzyme activities were measured. For the Italian agricultural soil I-BT, containing negligible amounts of organic pollutants but the highest Cu content, a Shannon index=2.13 was found. The enrichment of four mixed cultures capable of degrading solid phenanthrene in batch liquid systems was also studied. Phenanthrene degradation rates in batch systems were culture-dependent, and simple (one-slope) and complex (two-slope) kinetic behaviours were observed. The presence of common bands of microbial species in the cultures and in the native soil DNA indicated that those strains could be potential in situ phenanthrene degraders. Consistent with this assumption are the decrease of PAH and phenanthrene contents of Belgian soil B-BT and the isolation of phenanthrene-degrading bacteria. From the fastest phenanthrene-degrading culture C(B-BT), representative strains were identified as Achromobacter xylosoxidans (100%), Methylobacterium sp. (99%), Rhizobium galegae (99%), Rhodococcus aetherovorans (100%), Stenotrophomonas acidaminiphila (100%), Alcaligenes sp. (99%) and Aquamicrobium defluvium (100%). DGGE-profiles of culture C(B-BT) showed bands attributable to Rhodococcus, Achromobacter, Methylobacterium rhizobium, Alcaligenes and Aquamicrobium. The isolation of Rhodococcus aetherovorans and Methylobacterium sp. can be consistent with the hypothesis that different phenanthrene-degrading strategies, cell surface properties, or the presence of xenobiotic-specific membrane carriers could play a role in the uptake/degradation of solid phenanthrene.

Bacteria↗

Evaluation of pyrrolidonyl arylamidase for the identification of nonfermenting Gram-negative rods.

To evaluate the activity of pyrrolidonyl arylamidase (PYR) for the differentiation and identification of nonfermenting gram negative rods (NFGNR), 293 isolates were tested. A 24 h culture of each test organism was prepared. From this a 108-109 cfu/mL suspension was added to 0.25 mL of sterile physiologic solution. A PYR disk was then added and the test was incubated for 30 minutes at 35-37 degrees C, at environmental atmosphere. Reading was done by adding 1 drop of cinnamaldehyde reagent. Strains of Acinetobacter baumannii, Acinetobacter haemolyticus, Alcaligenes faecalis, Bergeyella zoohelcum, Bordetella bronchiseptica, Bordetella hinzii, Brevundimonas diminuta, Brevundimonas vesicularis, Brucella ovis, Brucella spp., Brucella suis, Burkholderia cepacia complex, Moraxella catarrhalis, Moraxella lacunata, Moraxella nonliquefaciens, Moraxella osloensis, Oligella ureolytica, Pseudomonas alcaligenes, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas Vb3, Psychrobacter phenylpyruvicus, and Stenotrophomonas maltophilia were PYR negative. On the other hand Achromobacter piechaudii, Achromobacter denitrificans, Achromobacter xylosoxidans, Burkholderia gladioli, Chryseobacterium gleum-indologenes, Comamonas testosroni, Cupriavidus pauculus, Delftia acidovorans, Elizabethkingia meningoseptica, Myroides spp., Ochrobactrum anthropi, Pseudomonas oryzihabitans, Ralstonia pickettii, Rhizobium radiobacter, Shewanella spp., Sphingobacterium multivorum, Sphingobacterium spiritivorum, and Weeksella virosa were PYR positive. Finally, Acinetobacter lwoffii, Pseudomonas aeruginosa, Pseudomonas fluorescens, Roseomonas spp., and Sphingomonas paucimobilis-parapaucimobilis were PYR variable. PYR testing should be considered as a useful tool to facilitate the identification of NFGNR.

Aminopeptidases↗