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PubMedDate not supplied

Infectious bursal disease virus and Alcaligenes faecalis infections in turkeys.

To determine if infectious bursal disease virus (IBDV) augments alcaligenes rhinotracheitis (ART), turkey poults were exposed to IBDV, Alcaligenes faecalis, or both IBDV and A. faecalis. In five experiments, poults exposed to IBDV alone exhibited neither signs of disease nor histopathologic lesions. Serum antibodies to IBDV were detected in poults exposed to this virus by inoculation and by direct contact with inoculated birds. Signs of ART were observed 4 to 6 days following exposure to A. faecalis. Clinical signs of ART and histopathologic lesions in the upper respiratory tract of poults exposed to both IBDV and A. faecalis were similar to those observed in poults exposed to A. faecalis alone.

Alcaligenes↗
PubMedDate not supplied

Evidence for adherence-dependent cytotoxicity of Alcaligenes faecalis in turkey tracheal organ cultures.

A virulent isolate of Alcaligenes faecalis was examined in turkey tracheal organ cultures (TTOC) for adherence using immunofluorescent staining and for cytotoxicity using light microscopic observation. Treatment of the bacterial culture with trypsin, antiserum specific for A. faecalis, and N-acetylneuraminic acid inhibited the ability of the organism to adhere to TTOC. Treatment of the bacterium with D-galactose partly decreased adherence of the bacterium. Those treatments that inhibited the adherence of A. faecalis also inhibited the cytolytic activity in TTOC. Treatment of the bacterial culture with D-galactose only partly decreased the cytolytic activity. These data indicate that adherence of the organism to TTOC is necessary for the cytolytic activity characteristic of A. faecalis isolates capable of causing alcaligenes rhinotracheitis in turkeys.

Adhesiveness↗
PubMedDate not supplied

Alcaligenes faecalis rhinotracheitis in Manitoba turkeys.

An outbreak of alcaligenes rhinotracheitis occurred on one premises housing five turkey flocks totaling 25,000 poults. Prominent findings were severe respiratory difficulty resulting from excess mucus in the nasopharynx, lachrimation, and tracheal collapse. Sinus and tracheal cultures consistently yielded Alcaligenes faecalis. An adenovirus was isolated and four flocks became positive for CELO virus by agar-gel-precipitin (AGP) tests. Mortality by flocks ranged from 4% to 48%. Treatment was unsuccessful and appeared to increase the mortality rate. The course of the disease was about 6 weeks, and recovered turkeys were marketed 1 week later than the usual date.

Alcaligenes↗
PubMedDate not supplied

Histamine-sensitizing factor of Alcaligenes faecalis.

Alcaligenes faecalis produced a histamine-sensitizing factor (HSF) in turkey poults and mice, which was detected in poults by an infraorbital sinus test and passive cutaneous anaphylaxis test and in mice by a rectal-temperature differential test. The A. faecalis HSF appeared to be similar to that produced by the genus Bordetella and may be partly responsible for the clinical signs of alcaligenes rhinotracheitis in young poults.

Adjuvants, Immunologic↗
PubMedDate not supplied

Control of turkey Alcaligenes rhinotracheitis in Utah with a live vaccine.

During the 1980 growing season in the Sanpete Valley of Utah, about half of the 250 to 300 flocks of turkeys were vaccinated with an oral vaccine against alcaligenes rhinotracheitis (ART). The vaccine consisted of a temperature-sensitive mutant of Alcaligenes faecalis. Most vaccinated birds developed serum antibodies. No outbreaks of ART occurred in vaccinated flocks, although some outbreaks occurred in unvaccinated flocks. During 1979, when on flocks were vaccinated, over 90% of the flocks experienced outbreaks of ART during late summer.

Alcaligenes↗
PubMedDate not supplied

Effect of humidity on infection of turkeys with Alcaligenes faecalis.

Turkeys maintained at 75% to 80% relative humidity were more adversely affected by Alcaligenes faecalis infection than turkeys maintained at 20 to 35% relative humidity. Alcaligenes faecalis was reisolated earlier and more often from turkeys maintained at the higher humidity. Clinically, the turkeys maintained at high humidity exhibited both sinusitis and conjunctivitis earlier than the turkeys at low humidity. In both groups, antibody titers as determined by a microagglutination test developed by 2 weeks postinoculation and started to decline after the third week, lymphocytosis was demonstrated at 1 week postinoculation, and a lymphopenia developed at 5 weeks postinoculation.

Agglutination Tests↗
PubMed1981

Cytotoxic activity of pathogenic Alcaligenes faecalis in turkey tracheal organ cultures.

Turkey tracheal organ cultures were used to study the virulence of Alcaligenes faecalis isolants that have been shown to be pathogenic for turkey poults. Viable infected and noninfected tracheal rings were examined by phase-contrast microscopy, and fixed stained sections were examined by light microscopy. Alcaligenes faecalis at concentrations of 10(8) and 10(9) colony-forming units/ml caused ciliostasis, hydropic degeneration (characterized by blebbing of the plasma membrane, cellular swelling, and cytoplasmic vacuolation), and eventual sloughing of the ciliated epithelium. Only ciliated epithelial cells appeared affected. For comparison, other bacterial isolants not pathogenic for turkeys were tried in this system. These bacterial isolants included 3 isolants of A faecalis from human beings and isolants of Klebsiella pneumoniae, Escherichia coli, and A faecalis from turkeys. Inoculation of each of these bacterial cultures onto tracheal organ cultures failed to produce the lesions described.

Alcaligenes↗
PubMedDate not supplied

[Attempt to establish genome similarity between Alcaligenes faecalis and several Gram-negative bacteria using the method of molecular hybridization of DNA to DNA].

The technique of DNA-DNA molecular hybridization was used to demonstrate the absence of genome similarity between Alcaligenes faecalis and gram-negative bacteria belonging to the genera Pseudomonas, Enterobacterium, Comamonas producing alkalis, and vibrions. Some similarity in the polynucleotide sequences was found between the DNA of the reper strain of Alcaligenes faecalis 45 (5--10% of homology) and the DNA of the culture belonging to the Achromobacter genus; therefore, the two genera may belong to one and the same family.

Alcaligenes↗
PubMedDate not supplied

FMN cofactor dissociation from the soluble hydrogenase of Alcaligenes eutrophus H16.

The specific activity of purified soluble hydrogenase of Alcaligenes eutrophus H16 was found to vary with enzyme concentration. Specific activity as a function of concentration of purified enzyme could be fit to an equation describing the dissociation of a compound into two components. An association constant, kappa(a), was determined in this way to be 39.4 +/- 8.7 micrograms protein/ml. The concentration of the enzyme affected its kinetic parameters: a tenfold decrease in enzyme concentration caused by a reduction of the V(max) and Kappa(m) (NAD) values to 45% and 58%, respectively, of the values for undiluted (0.64 mg/ml) enzyme. Diaphorase (NAD-dependent reduction of benzyl viologen) specific activity of the hydrogenase was unaffected by dilution. The extent of dilution-induced activity loss was dependent on pH, with greater activity loss observed at higher pH values. The substrate NAD prevented loss of specific activity due to dilution, while the product NADH did not. Specific activity loss due to dilution as reversed with the addition of the cofactor FMN. Dilution of the hydrogenase caused an increase in the enzyme's specific flavin fluorescence. These results suggest that dilution of the soluble hydrogenase of Alcaligenes eutrophus causes dissociation of the cofactor FMN, and this activity loss should be taken into account as an important factor governing hydrogenase activity and kinetic properties.

Alcaligenes↗
PubMed2003

Comparison of aspartate transcarbamoylase regulation in Pseudomonas alcaligenes and Pseudomonas mendocina.

The regulation of aspartate transcarbamoylase activity in cell extracts of Pseudomonas alcaligenes ATCC 14909 and Pseudomonas mendocina ATCC 25411 was compared. Under saturating substrate concentrations, pyrophosphate, CTP, UDP and ADP were highly inhibitory of the P. alcaligenes transcarbamoylase activity while pyrophosphate, UDP, ADP, ATP and GTP were the most effective inhibitors of the P. mendocina transcarbamoylase. By examining transcarbamoylase inhibition by ribonucleotide triphosphates, it was possible to differentiate these species assigned to different DNA homology groups and such an analysis might prove useful in the reclassification of Pseudomonas species.

Aspartate Carbamoyltransferase↗
PubMed1997

Characterization of the cryptic plasmids of the Pseudomonas alcaligenes type strain.

The species type strain of Pseudomonas alcaligenes contains three small cryptic plasmids (designated pECB1, 2, and 3) of 7740, 4480, and 2700 bp, respectively. Partial restriction enzyme maps have been constructed for pECB1 and 2 which on this basis do not appear to be related. pECB3 proved refractile to cutting with commonly used restriction enzymes, though it was completely rendered by those enzymes which recognize 4-bp sequences containing only G + C. This suggested that pECB3 is especially rich in these bases. Hybridization studies using labeled pECB2 as probe revealed homology with pECB3 and with regions of the bacterial chromosome, but not with pECB1. A 1214-bp region of pECB2 showed great sequence similarity to the basic replicon of pPS10, a 10-kb Pseudomonas-specific plasmid isolated from Pseudomonas syringae pv. savastonoi. The putative replicon (including the gene for a replication protein) was subcloned and both DNA strands were sequenced. Introduction of the putative replicon into the Escherichia coli plasmid pUC19 created a recombinant vector able to replicate in both E. coli and Pseudomonas spp. Minicell analyses did not reveal any peptides which could be attributed to the remaining region of pECB2 or to pECB1--a finding supported by sequencing studies. Attempts at plasmid curing were unsuccessful. A phenotypic comparison with a non-plasmid-harboring strain of P. alcaligenes, based on nutritional versatility and antibiotic susceptibility, revealed a single difference of note: the type strain alone was able to utilize benzoate for growth. Transformation of the non-harboring strain with pECB1-3, followed by selection on a minimal medium containing benzoate, gave no colonies. The advantages gained by possession of pECB1-3, if any, are at present unknown.

Amino Acid Sequence↗
PubMed1998

IS1491 from Pseudomonas alcaligenes NCIB 9867: characterization and distribution among Pseudomonas species.

A new insertion sequence, IS1491, has been cloned and sequenced. The 2489-bp IS1491 was isolated from a Pseudomonas alcaligenes NCIB 9867 (strain P25X) 4.8-kb PstI chromosomal fragment. IS1491 is flanked by an imperfect inverted repeat of 23 bp and carries two overlapping open reading frames, ORF1 and ORF2. Both ORF1 and ORF2 displayed homology to the IstA-like and IstB-like transposases encoded by the IS21 family of insertion sequences, which include two IS elements previously isolated from P. alcaligenes P25X, IS1474, and IS1475 (Yeo, C. C., and Poh, C. L. (1997). FEMS Microbiol. Lett. 149, 257-263). Transposition assays showed that IS1491 transposed at a frequency of approximately 1.4 x 10(-6). Transposition of IS1491 into the target pRK415 replicon was observed but when ORF2 was disrupted, a fusion between the donor and target replicons was detected. IS1491-like sequences were detected in total DNA of Pseudomonas putida NCIB 9869 (strain P35X), Pseudomonas aeruginosa, Pseudomonas stutzeri, Pseudomonas syringae, Pseudomonas mendocina, Comomonas acidovorans, and Comomonas testosteroni by hybridization with IS1491 DNA.

Amino Acid Sequence↗
PubMed1996

IS1394 from Pseudomonas alcaligenes N.C.I.B. 9867: identification and characterization of a member of the IS30 family of insertion elements.

A new insertion sequence designated IS1394 was isolated from Pseudomonas alcaligenes NCIB 9867 (P25X) by entrapment in plasmid pUCD800 which carries the Bacillus subtilis sacB and sacR genes. The 1100-bp sequence contains 27-bp inverted repeats with 4 bp mismatch and has one long open reading frame, spanning 92.1% of the entire IS. The deduced 338 amino-acid sequence demonstrated homology (varying from 65% to 78% similarity and 36-67% identity) to transposases encoded by the IS30 family of IS elements. Comparison of four different IS-sacB junction sequences showed that IS1394 generated 3-bp direct repeats of target DNA upon insertion. IS1394 is present in at least 10 copies in the P25X genome but none was detected in its endogenous plasmid pRA2. Hybridization experiments revealed that the distribution of IS1394 is limited to closely related strains, being present in three copies in Pseudomonas putida NCIB 9869 (P35X) and two copies in Pseudomonas alcaligenes ATCC type strain (ATCC 14904).

Base Sequence↗
PubMedDate not supplied

Characterization of an extracellular medium-chain-length poly(3-hydroxyalkanoate) depolymerase from Pseudomonas alcaligenes LB19.

An extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase from an isolate, Pseudomonas alcaligenes LB19, was purified to electrophoretic homogeneity by hydrophobic interaction chromatography using Octyl-Sepharose CL-4B and gel permeation chromatography using Sephadex G-150. The molecular mass of the enzyme, which consisted of a single polypeptide chain, was approximately 27.6 kDa. The pI value of the enzyme was estimated to be 5.7, and its maximum activity was observed at pH 9.0 and 45 degreesC. The enzyme was significantly inactivated by EDTA and phenylmethylsulfonyl fluoride (PMSF) but insensitive to dithiothreitol. It was also markedly inhibited by 0.1% Tween 80 and 0.05% Triton X-100. The purified enzyme could hydrolyze various types of bacterial aliphatic and aromatic MCL-PHAs but not poly(3-hydroxybutyrate), polycaprolactone, and poly(L-lactide). Biodegradation rates of the aromatic MCL-PHAs were significantly lower than those of the aliphatic MCL-PHAs, regardless of the compositions and types of aromatic substituents. It was able to hydrolyze medium-chain-length p-nitrophenylalkanoates more efficiently than the shorter-chain forms. The main hydrolysis products of poly(3-hydroxynonanoate) were identified as monomer units. The results demonstrated in this study suggest that the MCL-PHA depolymerase from P. alcaligenes LB19 is a distinct enzyme, which are different from those of other MCL-PHA degrading bacteria in its quaternary structure, pI value, sensitivity to EDTA and PMSF, and hydrolysis products of MCL-PHA.

Carboxylic Ester Hydrolases↗
PubMed1973

Studies of lipid A fractions from the lipopolysaccharides of Pseudomonas aeruginosa and Pseudomonas alcaligenes.

Lipid A fractions from Pseudomonas aeruginosa and Pseudomonas alcaligenes have similar compositions and structural features. By means of hydrazinolysis of the parent lipopolysaccharides and partial hydrolysis of the deacylation products, it was established that both lipids are derived from the beta-(1-->6)-linked disaccharide of glucosamine. Phosphorylated derivatives of the disaccharide from Ps. aeruginosa were also characterized. The lipids differ mainly in the absence of hexadecanoic acid and 2-hydroxydodecanoic acid from the lipid from Ps. alcaligenes. Evidence that in Ps. aeruginosa these acids are ester-linked to residues of 3-hydroxyalkanoic acids (including 3-hydroxydecanoic acid) was obtained. Heterogeneity of lipid A fractions was indicated by t.l.c., and by gel filtration of de-O-acylation products from mild alkaline methanolysis of the lipids.

Acylation↗
PubMed2003

Effect of carbon source on pyrimidine biosynthesis in Pseudomonas alcaligenes ATCC 14909.

The effect of carbon source on the regulation of the de novo pyrimidine biosynthetic enzymes in Pseudomonas alcaligenes ATCC 14909 was investigated. The de novo pyrimidine biosynthetic enzymes were measured in extracts of P. alcaligenes ATCC 14909 cells and of cells from an auxotroph deficient for orotate phosphoribosyltransferase activity. Pyrimidine biosynthetic enzyme activities in ATCC 14909 were influenced by pyrimidine supplementation to the culture medium but not by the carbon source present. Pyrimidine limitation of the auxotroph elevated the de novo enzyme activities indicating that this pathway may be controlled at the transcriptional level by a pyrimidine-related compound. Its regulation seemed to be subject to less transcriptional control by a pyrimidine-related compound than what was observed in the closely related species Pseudomonas pseudoalcaligenes.

Aspartate Carbamoyltransferase↗
PubMed1983

Pseudomonas alcaligenes endocarditis.

Pseudomonas alcaligenes is a common soil and water inhabitant that has rarely been proven a human pathogen. We describe a fatal case of Pseudomonas alcaligenes endocarditis. The need for accurate identification of unusual organisms isolated in a clinical setting are discussed.

Endocarditis, Bacterial↗
PubMed1995

Structure of a new azurin from the denitrifying bacterium Alcaligenes xylosoxidans at high resolution.

It has been reported previously that Alcaligenes xylosoxidans (NC1MB 11015) grown under denitrifying conditions produces two azurins instead of the single previously identified azurin [Dodd, Hasnain, Hunter, Abraham, Debenham, Kanzler, Eldridge, Eady, Ambler & Smith (1995). Biochemistry. In the press]. The new azurin, called azurin II, has been crystallized as blue elongated rectangular prisms with the tetragonal space group P4(1)22 and unit-cell parameters a = b = 52.65, c = 100.63 A. X-ray crystallographic data extending to 1.9 A resolution were collected by the Weissenberg method using 200 x 400 mm image plates and synchrotron X-rays of wavelength 0.97 A. The three-dimensional structure of azurin II has been solved by the molecular-replacement method using the structure of azurin from Alcaligenes denitrificans NCTC 8582 with which this new azurin shows a close homology. The quality of the initial map was sufficient to predict a number of sequence differences. The model is currently refined to an R-factor of 18.8% with X-ray data between 8.5 and 1.9 A. The final model of 961 protein atoms, one Cu atom and 50 water molecules has r.m.s. deviations from ideality of 0.009 A for bond lengths and 1.7 degrees for bond angles. The overall structure is similar to that of the azurin from A. denitrificans NCTC 8582. It has a beta-barrel structure with the Cu atom located near the top end of the molecule. The Cu atom is coordinated to Ndelta of His46 and His117 at 2.02 A and to Sgamma of Cys112 at 2.12 A, while the carbonyl O atom of Gly45 and Sdelta atom of Met121 provide the additional interactions at 2.75 and 3.26 A, respectively.

Journal Article↗