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PubMed2005

Biodegradation of fenoxaprop-p-ethyl by bacteria isolated from sludge.

A mixed bacterial population was isolated using enrichment in a basal medium containing increasing amounts of fenoxaprop-p-ethyl as a sole carbon source from sludge that had been exposed to fenoxaprop-p-ethyl production wastewater for about 2 years. Eight kinds of isolates could utilize fenoxaprop-p-ethyl, but only one was identified belonging to genus Alcaligenes, named Alcaligenes sp. H. In pure culture, there was 45.8, 66.0 and 69.5% loss of fenoxaprop-p-ethyl (initial concentration: 100, 50, 25 ppm, respectively) as the sole carbon source with biodegradation by Alcaligenes sp. H and fenoxaprop-p-ethyl degradation kinetics obeyed the first-order kinetics, the same as the fenoxaprop-p-ethyl biodegradation kinetics in soil. At least five degradation products of fenoxaprop-p-ethyl biodegradation by Alcaligenes sp. H and two degradation products of fenoxaprop-p-ethyl biodegradation by Huv separated by HPTLC. It is possible that the fenoxaprop-p-ethyl biodegradation by Alcaligenes sp. H includes the same pathway as that by Huv comparing with the Rf.

Bacteria↗
PubMedDate not supplied

[Preparation of bacteria--degraders of alkylnaphthalenesulfonate by a molecular breeding method].

Transconjugant strain of bacteria degraders of alkylnaphthalenesulphonate(ANS)--P Pseudomonas alcaligenes TR (NPL-41) was obtained under conditions of ANS selective pressure and flow cultivation of bacteria destructors of alkylbenzene sulphonate (ABS) P. alcaligenes TR (pABS) and naphthalene P. putida BS438 (NPL-41) and their immobilization on an inert carrier. Transconjugant P. alcaligenes TR(NPL-41) was prepared by means of conjugative transfer of plasmid NPL-41 from P. putida BS438 to P. alcaligenes TR. The check out of transconjugant under conditions of flow cultivation showed their ability to degradation of ANS. Investigation of the stability of plasmid NPL-41 in the transconjugant P. alcaligenes TR(NPL-41), obtained by molecular breeding under conditions of flow cultivation and by conjugative transfer, showed the transconjugant prepared by the former method to be more stable.

Base Sequence↗
PubMed1977

Micromorphology of Gram-negative hydrogen bacteria. I. Cell morphology and flagellation.

The cell morphology, the arrangement and fine structure of flagella and the piliation of the following Gram-negative aerobic hydrogen bacteria have been studied: Alcaligenes eutrophus, Alcaligenes paradoxus, Alcaligenes ruhlandii, Pseudomonas flava, Pseudomonas pseudoflava, Pseudomonas palleronii, Pseudomonas facilis, Aquaspirillum autotrophicum, Paracoccus denitrificans, Corynebacterium autotrophicum, and strains MA 2 and SA 35. The identity of the bacteria was examined by their substrate spectra and type of flagellation. Three types of flagellar fine structure were differentiated. The presence of pili was noted in strains of Alcaligenes paradoxus, Pseudomonas flava, P.pseudoflava, P.palleronii, and P.facilis.

Alcaligenes↗
PubMed1987

Identification of dimethyl disulfide-forming bacteria isolated from activated sludge.

Twenty-four strains with high dimethyl disulfide (DMDS)-forming ability were isolated from activated sludge and identified to the genus level. These bacteria were classified into four groups (A, B, C, and D) by the API ZYM System (API System S.A., Montalieu, France). Group A (three strains) was identified as genus Lactobacillus by the API 20B System, by the method of Cowan and Steel, and by production of lactic acid as confirmed by gas-liquid chromatography. Group B (eight strains) was identified as genus Corynebacterium by API 20B and the Cowan and Steel method. Group C (one strain) was suggested to belong to genus Corynebacterium by the API 20B System. Group D (12 strains) was identified as genus Pseudomonas or Alcaligenes by the API 20B System, as genus Alcaligenes by the Cowan and Steel method, and as Achromobacter group Vd by the API 20NE System. However, on the basis of guanine-plus-cytosine contents in DNA and form of flagella, these strains were identified as genus Pseudomonas. Formation of DMDS from DL-methionine and S-methyl-L-cysteine was tested. DMDS-forming bacteria isolated from activated sludge formed DMDS from both precursors. In genus Pseudomonas, P. aeruginosa could not form DMDS from either precursor, but P. acidovorans, P. alcaligenes, P. pseudoalcaligenes, and P. testosteroni formed DMDS. In genus Alcaligenes, A. denitrificans subsp. xylosoxydans, A. denitrificans subsp. denitrificans, A. faecalis, and A. odorans formed DMDS from both precursors. Achromobacter group Vd formed DMDS from S-methyl-L-cysteine, but could not from DL-methionine.

Alcaligenes↗
PubMed2005

Surface active properties of bacterial strains isolated from petroleum hydrocarbon-bioremediated soil.

Two bacterial strains identified as Ralstonia picketti (BP 20) and Alcaligenes piechaudii (CZOR L-1B) were isolated from petroleum hydrocarbon-contaminated soil following bioremediation treatment. The surface active properties, e.g. surface tension, emulsification and foamability of their culture filtrates were evaluated. Bacterial cell-surface hydrophobicity (BAH) as measured by analyzing cell affinity towards aliphatic and aromatic compounds was also determinated. The bacteria grew in liquid cultures containing 1% (v/v) of crude oil as carbon and energy source at 30 degrees C under aerobic conditions. The surface tensions were reduced to 61 mN/m and 55 mN/m by Ralstonia picketti and Alcaligenes piechaudii, respectively. The emulsification index (EI24) was almost 100% for all tested compounds except diesel oil. The stability of the emulsions was deteminated at 4 degrees C, 45 degrees C and 65 degrees C. The emulsions were stable at 4 degrees C. Ralstonia picketti was better foam inducer (FV = 50 ml) compared to Alcaligenes piechaudii (FV = 10 ml). The BAH measurements revealed higher adhesion of Alcaligenes piechaudii cells towards different hydrocarbons compared to Ralstonia picketti cells. The strains were found to have a surface hydrophobicity in the following order: aliphatic hydrocarbons, BTEX, and PAHs. The ability to adhere to bulk hydrocarbon is mostly a characteristic of hydrocarbon-degrading bacteria. The strains were found to be better emulsifiers than surface tension reducers. They produce water-soluble extracellular bioemulsifiers. Both bacterial isolates have good properties to use them, mainly in the petroleum industry, e.g. in enhanced oil recovery and in bioremediation processes-primarily due to their emulsification property, i.e. emulsion forming and stabilizing capacity.

Alcaligenes↗
PubMed2003

Influence of growth phase on bacterial cell electrokinetic characteristics examined by soft particle electrophoresis theory.

The influence of incubation time on the electrokinetic properties of Escherichia coli, Pseudomonas putida, Alcaligenes faecalis, and Alcaligenes sp., was examined by electrophoretic mobility measurements and the results were discussed based on Ohshima's soft particle theory. The electrophoretic mobility of E. coli plotted against incubation time revealed that the mobility gradually increased from the outset of incubation to 7 h, which corresponded to the exponential growth and early stationary phase, and then decreased. For all strains, electrophoretic mobility leveled off to nonzero values in accordance with the increase in ionic concentration, which was a characteristic feature of soft particles. Soft particle analysis was carried out at the outset of incubation and the time when mobility reached a maximum in order to obtain the spatial charge density (ZN) and cell surface softness (1/lambda). ZN at the maximum electrophoretic mobility was more negative than that at the outset of incubation for E. coli, P. putida, and A. faecalis, while the ZN of Alcaligenes sp. almost remained unchanged. 1/lambda decreased for E. coli, but increased for Alcaligenes sp. and A. faecalis. These findings indicated that the cell growth phase affects both ZN and 1/lambda, although the dependencies of these parameters are unique to each bacterial strain.

Journal Article↗
PubMed1990

The aerobic psychotrophic populations on meat and meat contact surfaces in a meat production system and on meat stored at chill temperatures.

At a city abattoir, a wholesaler and 10 different supermarkets, surface microbiological samples were taken of carcasses, hands and apron fronts of members of staff and equipment (mincers and saws). In addition, minced meat, packaged and displayed in chilled cabinets, was also sampled. Carcasses, personnel surfaces and equipment were monitored by a modified agar sausage technique. From each of the highest dilution psychotrophic plate counts, five colonies were selected randomly, isolated and identified (1265 in total). Microbes developing on chilled meat were also isolated from other surfaces in the production chain. On chilled meat (51%) and at the abattoir (36%) pseudomonads were the predominant organisms followed by the Gram-positive cocci on chilled meat and by Acinetobacter, Moraxella and Alcaligenes spp. at the abattoir. At the wholesaler Gram-positive cocci (32%) predominated, followed by Alcaligenes, Moraxella and Alcaligenes spp. Pseudomonas, Alcaligenes, Neisseriaceae and related genera, Gram-positive cocci, species from the coryneform groups of bacteria and yeasts were identified from all the surfaces monitored. Identification with the API NE20 was unsatisfactory. Enterbacteriaceae, lactobacilli and endospore-forming bacteria were identified occasionally, but their significance as contaminating organisms seems low. No Salmonella spp. were identified.

Abattoirs↗
PubMed1992

Evaluation of the 4-hour RapID NF Plus method for identification of 345 gram-negative nonfermentative rods.

The ability of the RapID NF Plus system (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) to identify 345 nonfermentative gram-negative rods was evaluated. Kits were inoculated with no. 1 McFarland suspensions, and reactions were interpreted after a 4-h incubation at 35 degrees C. Overall, the method correctly identified 311 strains (90.1%) without additional tests and 21 strains (6.1%) with additional tests, and 13 strains (3.8%) were misidentified. Five of 13 misidentified strains were Alcaligenes faecalis-Alcaligenes odorans misidentified as Alcaligenes xylosoxidans; however, all strains were xylose negative but nitrate positive and could have been A. faecalis group I-Alcaligenes piechaudii. The system does not differentiate between Pseudomonas fluorescens and Pseudomonas putida, and all Acinetobacter species are identified as Acetinobacter calcoaceticus. Additionally, no subspecies differentiation is made between A. xylosoxidans subsp. xylosoxidans and A. xylosoxidans subsp. denitrificans. All strains of the former Flavobacterium group IIb are identified as Flavobacterium indologenes-Flavobacterium gleum, and no species identification of the genus Methylobacterium is attempted. The system is easy to set up and interpret and provides an accurate commercial nonautomated method for same-day identification of gram-negative nonfermenters.

Bacteriological Techniques↗
PubMed1999

Capillary electrophoresis-single-strand conformation polymorphism analysis for rapid identification of Pseudomonas aeruginosa and other gram-negative nonfermenting bacilli recovered from patients with cystic fibrosis.

We used capillary electrophoresis-single-strand conformation polymorphism (CE-SSCP) analysis of PCR-amplified 16S rRNA gene fragments for rapid identification of Pseudomonas aeruginosa and other gram-negative nonfermenting bacilli isolated from patients with cystic fibrosis (CF). Target sequences were amplified by using forward and reverse primers labeled with various fluorescent dyes. The labeled PCR products were denatured by heating and separated by capillary gel electrophoresis with an automated DNA sequencer. Data were analyzed with GeneScan 672 software. This program made it possible to control lane-to-lane variability by standardizing the peak positions relative to internal DNA size markers. Thirty-four reference strains belonging to the genera Pseudomonas, Brevundimonas, Burkholderia, Comamonas, Ralstonia, Stenotrophomonas, and Alcaligenes were tested with primer sets spanning 16S rRNA gene regions with various degrees of polymorphism. The best results were obtained with the primer set P11P-P13P, which spans a moderately polymorphic region (Escherichia coli 16S rRNA positions 1173 to 1389 [M. N. Widjojoatmodjo, A. C. Fluit, and J. Verhoef, J. Clin. Microbiol. 32:3002-3007, 1994]). This primer set differentiated the main CF pathogens from closely related species but did not distinguish P. aeruginosa from Pseudomonas alcaligenes-Pseudomonas pseudoalcaligenes and Alcaligenes xylosoxidans from Alcaligenes denitrificans. Two hundred seven CF clinical isolates (153 of P. aeruginosa, 26 of Stenotrophomonas maltophilia, 15 of Burkholderia spp., and 13 of A. xylosoxidans) were tested with P11P-P13P. The CE-SSCP patterns obtained were identical to those for the corresponding reference strains. Fluorescence-based CE-SSCP analysis is simple to use, gives highly reproducible results, and makes it possible to analyze a large number of strains. This approach is suited for the rapid identification of the main gram-negative nonfermenting bacilli encountered in CF.

Cystic Fibrosis↗
PubMed1985

A comparative study of the resonance Raman spectra of bacterial cytochromes.

Resonance Raman spectra were obtained for two newly isolated bacterial cytochromes, Alcaligenes faecalis (ATCC 8750) c554 and Alcaligenes faecalis c556. Their spectra were compared with those of mammalian cytochrome c and two other bacterial cytochromes, Paracoccus denitrificans c550 and Pseudomonas aeruginosa c551. The positions of the Raman bands indicated that, while Al. c554 and Al. c556 were c-type cytochromes with two thioether linkages, several common features found in their Raman spectra were anomalous. These features suggest that the electronic charge density of the porphyrin macrocycle of Al. c554 and Al. c556 is more asymmetric than that of other bacterial and mammalian c-type cytochromes. The Raman evidence indicates that the electronic properties of the heme are controlled by the protein in these two Alcaligenes cytochromes.

Alcaligenes↗
PubMed1993

Microbial synthesis of 3-deoxy-D-erythro-hex-2-ulosonic acid 6-phosphate.

A microbial route was explored for the synthesis of 3-deoxy-D-erythro-hex-2-ulosonic acid 6-phosphate (2-keto-3-deoxy-6-phosphogluconate, KDPG). Two strains of bacteria, Alcaligenes eutrophus H16 F34 (DSM 529) and Escherichia coli DF 71 (CGSC 4880), lacking in KDPG-aldolase activity were tested for excretion of KDPG. Using pyruvate and gluconate as carbon sources, Alcaligenes eutrophus H16 F34 accumulated and excreted 3-deoxy-D-erythro-hexulosonic acid 6-phosphate into the culture broth, while the E. coli strain, using pyruvate and glucuronate, failed. KDPG was isolated from the culture supernatant of Alcaligenes eutrophus H16 F34 in 78% yield and 5 g scale with respect to the consumed gluconate.

Alcaligenes↗
PubMed1984

Isolation and characterization of megaplasmid DNA from lithoautotrophic bacteria.

A method is described for the preparative isolation of megaplasmids ranging in size from 340 to 700 kb. These plasmids were isolated from chemolithoautotrophic bacteria including the species Alcaligenes, Pseudomonas, and Paracoccus. The procedure was based on alkaline sodium dodecyl sulfate lysis of the cells, followed by heat treatment, salt precipitation, several phenol extractions, dialysis steps, and proteinase and RNase treatment. The various parameters were evaluated and controlled. Hydrogen-oxidizing-ability (Hox) encoding plasmids were compared by EcoRI restriction enzyme analysis. pHG plasmids from Alcaligenes eutrophus wild-type strains appeared to be closely related; plasmids derived from the type strain TF93 and from A. hydrogenophilus exhibited major differences in restriction sites. Two cryptic plasmids harbored by Pseudomonas facilis and Paracoccus denitrificans showed scarcely detectable similarity to the plasmid species of Alcaligenes.

Alcaligenes↗
PubMed2005

The use of ozone in the remediation of polycyclic aromatic hydrocarbon contaminated soil.

The potential of using ozone for the removal of phenanthrene from several different soils, both alone and in combination with biodegradation using a microbial inoculant (Pseudomonas alcaligenes PA-10), was examined. The greater the water content of the soil the less effective the ozone treatment, with air-dried soils showing the greatest removal of phenanthrene; while soils with higher levels of clay also reduced the effectiveness of the ozone treatments. However, at least a 50% reduction in phenanthrene levels was achieved in air-dried soil after an ozone treatment of 6 h at 20 ppm, with up to 85% removal of phenanthrene achieved in sandy soils. The biodegradation results indicate that P. alcaligenes PA-10 may be useful as an inoculant for the removal of PAHs from contaminated soils. Under the conditions used in our experiments, however, pre-ozonation did not enhance subsequent biodegradation of phenanthrene in the soils. Similar levels of phenanthrene removal occurred in both non-ozonated and ozonated Cruden Bay soil inoculated with P. alcaligenes PA-10. However, the biodegradation of phenanthrene in ozonated Boyndie soil was much slower. This may be due to the release of toxic products in this soil during ozonation.

Biodegradation, Environmental↗
PubMed1982

The bacterial biogenesis of isobutyraldoxime O-methyl ether, a novel volatile secondary metabolite.

Production of the volatile metabolite, isobutyraldoxime O-methyl ether (IBME) by a Moraxella-like organism NCIB 11650 was investigated under a variety of environmental conditions using gas chromatography. Under aerobic conditions up to 10 micrograms IBME ml-1 was produced on mineral salts media containing 0.5% (w/v) glucose or succinate as sole C source with 0.1% (w/v) NH4Cl as sole N source. Exogenous L-valine further stimulated IBME formation up to 25 micrograms ml-1 but supplementation of the medium with D-isomer or other amino acids had little effect on IBME production and did not lead to the appearance of analogues of IBME. Trapping experiments using [14C]valine confirmed that IBME was derived from this amino acid. Several other bacterial species examined, e.g. Alcaligenes sp. NCIB 11652, another Moraxella-like organism NCIB 11651 and Pseudomonas sp. NCIB 11653 also produced IBME under similar conditions. The Alcaligenes strain synthesized up to 20 micrograms ml-1 in the absence of valine and up to 90 micrograms ml-1 in its presence. The product of IBME exhibited many features characteristic of the formation of a secondary metabolite. Thus biosynthesis was confined to a narrower range of temperature than cell division, was almost completely suppressed by 300 mM-phosphate and was inhibited by high concentrations of readily utilizable C sources. Although IBME synthesis in the Moraxella-like organism NCIB 11650 appeared to be growth-related, its formation by both the Alcaligenes sp. and the Moraxella-like organism NCIB 11651 was delayed until the late-exponential and early-stationary phases of growth. The biological significance of this novel class of secondary metabolite is discussed and a possible biosynthetic route proposed.

Alcaligenes↗
PubMed1989

Evidence for a new pathway in the bacterial degradation of 4-fluorobenzoate.

Six bacterial strains able to use 4-fluorobenzoic acid as their sole source of carbon and energy were isolated by selective enrichment from various water and soil samples from the Stuttgart area. According to their responses in biochemical and morphological tests, the organisms were assigned to the genera Alcaligenes, Pseudomonas, and Aureobacterium. To elucidate the degradation pathway of 4-fluorobenzoate, metabolic intermediates were identified. Five gram-negative isolates degraded this substrate via 4-fluorocatechol, as described in previous studies. In growth experiments, these strains excreted 50 to 90% of the fluoride from fluorobenzoate. Alcaligenes sp. strains RHO21 and RHO22 used all three isomers of monofluorobenzoate. Alcaligenes sp. strain RHO22 also grew on 4-chlorobenzoate. Aureobacterium sp. strain RHO25 transiently excreted 4-hydroxybenzoate into the culture medium during growth on 4-fluorobenzoate, and stoichiometric amounts of fluoride were released. In cell extracts from this strain, the enzymes for the conversion of 4-fluorobenzoate, 4-hydroxybenzoate, and 3,4-dihydroxybenzoate could be detected. All these enzymes were inducible by 4-fluorobenzoate. These data suggest a new pathway for the degradation of 4-fluorobenzoate by Aureobacterium sp. strain RHO25 via 4-hydroxybenzoate and 3,4-dihydroxybenzoate.

Alcaligenes↗
PubMed1998

Development of PCR primer systems for amplification of nitrite reductase genes (nirK and nirS) to detect denitrifying bacteria in environmental samples.

A system was developed for the detection of denitrifying bacteria by the amplification of specific nitrite reductase gene fragments with PCR. Primer sequences were found for the amplification of fragments from both nitrite reductase genes (nirK and nirS) after comparative sequence analysis. Whenever amplification was tried with these primers, the known nir type of denitrifying laboratory cultures could be confirmed. Likewise, the method allowed a determination of the nir type of five laboratory strains. The nirK gene could be amplified from Blastobacter denitrificans, Alcaligenes xylosoxidans, and Alcaligenes sp. (DSM 30128); the nirS gene was amplified from Alcaligenes eutrophus DSM 530 and from the denitrifying isolate IFAM 3698. For each of the two genes, at least one primer combination amplified successfully for all of the test strains. Specific amplification products were not obtained with nondenitrifying bacteria or with strains of the other nir type. The specificity of the amplified products was confirmed by subsequent sequencing. These results suggest the suitability of the method for the qualitative detection of denitrifying bacteria in environmental samples. This was shown by applying one generally amplifying primer combination for each nir gene developed in this study to total DNA preparations from aquatic habitats.

Alcaligenes↗
PubMed1966

Factors influencing the occurrence of high numbers of iodine-resistant bacteria in iodinated swimming pools.

It has been shown that, although iodinated swimming-pool waters are usually free from coliform bacteria and enterococci, the total counts frequently become relatively high. Pseudomonas alcaligenes and Alcaligenes faecalis have been shown to account for most of these high counts. It was of interest, therefore, to compare the microbial flora of four alternately chlorinated and iodinated swimming pools. By means of the membrane filter method and suitable selective media, examinations were made for total viable counts, coliform bacteria, enterococci, staphylococci, Streptococcus salivarius, and P. aeruginosa. Colonies also were picked from membrane filters incubated on standard plate count agar and identified. The results showed that, although viable counts were significantly higher during the iodinated periods, the specific types of bacteria determined were either fewer than or the same as in chlorinated periods. During chlorination, the predominant microbial flora consisted of staphylococci and members of the genus Bacillus. During iodination, however, the P. alcaligenes-A. faecalis group accounted for 92 to 99% of the microbial flora. The accumulation of high numbers of these bacteria was shown to be due to their iodine resistance and their ability to grow rapidly in pool water in the absence of free iodine.

Alcaligenes↗
PubMed1990

Different types of dienelactone hydrolase in 4-fluorobenzoate-utilizing bacteria.

Of various benzoate-utilizing bacteria tested, Alcaligenes eutrophus 335, A. eutrophus H16, A. eutrophus JMP222, A. eutrophus JMP134, Alcaligenes strain A7, and Pseudomonas cepacia were able to grow with 4-fluorobenzoate as the sole source of carbon and energy. P. cepacia also utilizes 3-fluorobenzoate. Except for A. eutrophus JMP134, which is known to grow with 2,4-dichlorophenoxyacetate and 3-chlorobenzoate (R. H. Don and J. M. Pemberton, J. Bacteriol. 145:681-686, 1981), the strains were unable to grow at the expense of these compounds or 4-chlorobenzoate. Assays of cell extracts revealed that all strains express dienelactone hydrolase and maleylacetate reductase activities in addition to enzymes of the catechol branch of the 3-oxoadipate pathway when growing with 4-fluorobenzoate. Induction of dienelactone hydrolase and maleylacetate reductase apparently is not necessarily connected to synthesis of catechol 1,2-dioxygenase type II and chloromuconate cycloisomerase activities, which are indispensable for the degradation of chlorocatechols. Substrate specificities of the dienelactone hydrolases provisionally differentiate among three types of this activity. (i) Extracts of A. eutrophus 335, A. eutrophus H16, A. eutrophus JMP222, and Alcaligenes strain A7 convert trans-4-carboxymethylenebut-2-en-4-olide (trans-dienelactone) much faster than the cis-isomer (type I). (ii) The enzyme present in P. cepacia shows the opposite preference for the isomeric substrates (type II). (iii) Cell extracts of A. eutrophus JMP134, as well as purified dienelactone hydrolase from Pseudomonas strain B13 (E. Schmidt and H.-J. Knackmuss, Biochem. J. 192:339-347, 1980), hydrolyze both dienelactones at rates that are of the same order of magnitude (type III). This classification implies that A. eutrophus JMP134 possesses at least two different dienelactone hydrolases, one of type III encoded by the plasmid pJP4 and one of type I, which is also present in the cured strain JMP222.

Alcaligenes↗