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At least 19 recordsLinked to original sources
PubMed1992

Specificity of the Pseudomonas aeruginosa PAO1 lipoprotein I gene as a DNA probe and PCR target region within the Pseudomonadaceae.

The lipoprotein I gene (oprI) of Pseudomonas aeruginosa PAO1 was cloned and sequenced. A high degree of homology was found between our cloned PAO1 gene sequence and two published oprI sequences. Specific oligonucleotides were designed to amplify the oprI gene by the polymerase chain reaction (PCR). The potential of either the complete gene sequence or the specific oligonucleotide primers as a tool for rapid strain identification was directly assessed against bacterial colonies by PCR or against purified genomic DNA by Southern blot analysis, using a number of representative strains within the Pseudomonadaceae. The oprI gene was found to be well conserved within RNA group I.

Amino Acid Sequence↗
PubMed1990

Prevalence and antimicrobial susceptibility of Enterobacteriaceae, Pseudomonadaceae and Acinetobacter in human periodontitis.

This study examined the occurrence of non-oral gram-negative facultatively anaerobic rods in advanced adult periodontitis. Speciation and in vitro antimicrobial susceptibility testing was performed using the MicroScan micromethod system. A total of 42 taxa of Enterobacteriaceae, Pseudomonadaceae and Acinetobacter were isolated from 427 of 3,050 (14.0%) patients. In 159 (5.2%) patients, these organisms comprised more than 5% of the cultivable subgingival microflora. Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Klebsiella oxytoca and Enterobacter agglomerans accounted for more than 50% of all strains isolated. Virtually all study strains demonstrated high in vitro susceptibility to ciprofloxacin, but exhibited variable susceptibility patterns to 18 other antimicrobial agents tested. In 3 "refractory" periodontitis patients heavily infected with enteric rods, systemic ciprofloxacin therapy (500 mg BiD for 10 days) led to resolution of the subgingival infections and improved clinical periodontal status. The present findings indicate that 5% of severe periodontitis lesions may harbor high levels of non-oral, gram-negative, facultatively anaerobic rods. Systemic ciprofloxacin appears to be capable of eradicating these potential pathogens from deep periodontal pockets.

Acinetobacter↗
PubMed2006

Identification and specific detection of a novel pseudomonadaceae cluster associated with soils from winter wheat plots of a long-term agricultural field experiment.

The genus Pseudomonas (sensu stricto) represents a group of microorganisms directly involved in functions conferring plant health. We performed a study in the DOK long-term agricultural field experiment on the basis of previously published Pseudomonas-selective PCR primers in order to investigate the community structure of the microbial groups defined by the target range of these primers. Three different agricultural management systems, i.e., conventional, biodynamic, and bio-organic, along with mineral and unfertilized controls were investigated, with each system planted with either winter wheat or a grass-clover ley. Amplified small-subunit rRNA gene fragments were analyzed using the genetic profiling techniques restriction fragment length polymorphism (RFLP) and denaturing gradient gel electrophoresis (DGGE), revealing distinct differences between soils planted with winter wheat and grass clover but only minor differences between the management systems. Phylogenetic analyses of 59 clone sequences retrieved from bio-organic and unfertilized systems identified sequences related to Pseudomonas fluorescens and a novel cluster termed Cellvibrio-related Pseudomonadaceae (CRP). The CRP clones were exclusively isolated from winter wheat soil samples and were responsible for the crop-specific differences observed in RFLP and DGGE profiles. New primers were designed for the amplification of CRP targets directly from soil DNA, yielding strong signals exclusively for winter wheat soils. We concluded that crop-associated CRP exist in agricultural soils and that genetic profiling followed by specific probe design represents a valuable approach for identification as well as sensitive and rapid monitoring of novel microbial groups in the environment.

Agriculture↗
PubMed1991

Conservation of the gene for outer membrane protein OprF in the family Pseudomonadaceae: sequence of the Pseudomonas syringae oprF gene.

The conservation of the oprF gene for the major outer membrane protein OprF was determined by restriction mapping and Southern blot hybridization with the Pseudomonas aeruginosa oprF gene as a probe. The restriction map was highly conserved among 16 of the 17 serotype strains and 42 clinical isolates of P. aeruginosa. Only the serotype 12 isolate and one clinical isolate showed small differences in restriction pattern. Southern probing of PstI chromosomal digests of 14 species from the family Pseudomonadaceae revealed that only the nine members of rRNA homology group I hybridized with the oprF gene. To reveal the actual extent of homology, the oprF gene and its product were characterized in Pseudomonas syringae. Nine strains of P. syringae from seven different pathovars hybridized with the P. aeruginosa gene to produce five different but related restriction maps. All produced an OprF protein in their outer membranes with the same apparent molecular weight as that of P.aeruginosa OprF. In each case the protein reacted with monoclonal antibody MA4-10 and was similarly heat and 2-mercaptoethanol modifiable. The purified OprF protein of the type strain P. syringae pv. syringae ATCC 19310 reconstituted small channels in lipid bilayer membranes. The oprF gene from this latter strain was cloned and sequenced. Despite the low level of DNA hybridization between P. aeruginosa and P. syringae DNA, the OprF gene was highly conserved between the species with 72% DNA sequence identity and 68% amino acid sequence identity overall. The carboxy terminus-encoding region of P. syringae oprF showed 85 and 33% identity, respectively, with the same regions of the P. aeruginosa oprF and Escherichia coli ompA genes.

Amino Acid Sequence↗
PubMed1989

[Study of the inhibition of neutral phosphatase from Pseudomonadaceae by derivatives of its substrates].

The inhibition of neutral phosphatase isolated from the bacteria of the Pseudomonadaceae family by various fragments of the enzyme-hydrolyzed R-O-PO3H2 substrates, inorganic orthophosphate (KH2PO4) and its analogs as well as by adenine, adenosine, alcohols, sugars and amino acids, was studied. It was demonstrated that among other compounds tested only the orthophosphoric acid anions (H2PO4-) exhibit the properties of strong associative inhibitors (K1Vi = 4.35.10(-6)M of the enzyme. The pH dependence of the Michaelis constant [pKm0 = f(pH)] and the inhibition constant for phosphatase by potassium orthophosphate [pK1Vi(KH2PO4) = f(pH)] was studied. The presence in the enzyme active center of a carboxylic (pK = 4.3 +/- 0.1) (presumably, glutamine) and an imidazole (pK = 7.15 +/- 0.1) amino acid residues was postulated. The data obtained were compared to those for neutral, alkaline and acid phosphatases.

Alcohols↗
PubMed1981

[Quantitative and species composition of conditionally-pathogenic microorganisms of the families Enterobacteriaceae and Pseudomonadaceae in healthy subjects and intestinal dysbacteriosis].

The spread and quantitative content of opportunistic microorganisms belonging to the families Enterobacteriaceae and Pseudomonadaceae in clinically healthy persons and in somatic patients with intestinal dysbacteriosis were studied. Opportunistic enterobacteria were shown to be widely spread in healthy persons (K. pneumoniae in 45%, Citrobacter in 44%, E. aerogenes in 17%, E. cloaceae in 11%). In clinically healthy persons most of these cultures (72.3%) were detected in an amount not exceeding 1 X 10(4) organisms per 1 g of feces, while in patients with intestinal dysbacteriosis opportunistic enterobacteria were detected in most cases (70.9%) in an amount of 1 X 10(5)--1 X 10(8) organisms per 1 g of feces. The results of this study suggest that the presence of the opportunistic enterobacteria K. pneumoniae, Citrobacter, E. aerogenes and E. cloaceae in an amount not exceeding 1 X 10(4) organisms per 1 g of feces is normal for healthy persons, while the increase of their presence in the intestine to 1 X 10(6)--1 X 10(8) is indicative of intestinal dysbacteriosis.

Abdomen↗
PubMedDate not supplied

[Features of secretion of bacteriolytic enzymes and polysaccharides in bacteria from the Pseudomonadaceae family].

It is shown that secretion of bacteriolytic complex enzymes in a bacterium of the family Pseudomonadaceae begins in the latent and early logarithmic culture growth phases. A maximum of specific bacteriolytic activity falls in the same time interval with a maximum of specific activity of lytic proteinase L2 and muramidase. The activity of concomitant enzymes--non-specific proteinase and neutral phosphatase--peaks in the middle of the logarithmic growth phase. Secretion of exopolysaccharide into the external medium begins in the second half of the logarithmic growth phase and, thus, is not coordinated with secretion of bacteriolytic enzymes. It is shown that relative content of enzymes and polysaccharides depends upon the composition of growth medium for the producer.

Culture Media↗
PubMed1994

Cloning and nucleotide sequence of the Pseudomonas aeruginosa glucose-selective OprB porin gene and distribution of OprB within the family Pseudomonadaceae.

OprB is a glucose-selective porin known to be produced by Pseudomonas aeruginosa and Pseudomonas putida. We have cloned and sequenced the oprB gene of P. aeruginosa and obtained expression of OprB in Escherichia coli. The mature protein consists of 423 amino acid residues with a deduced molecular mass of 47597 Da. Several clusters of amino acid residues, potentially involved in the structure or function of the protein, were identified. An area of regional homology with E. coli LamB was also identified. Carbohydrate-inducible proteins, potentially homologous to OprB, were identified in several rRNA homology-group-I pseudomonads by sodium dodecyl sulfate/polyacrylamide gel electrophoresis analysis, Western immunoblotting and N-terminal amino acid sequencing. These species also contained DNA that hybridized to a P. aeruginosa oprB gene probe.

Amino Acid Sequence↗
PubMed1980

Variable enzymological patterning in tyrosine biosynthesis as a means of determining natural relatedness among the Pseudomonadaceae.

Enzymes of tyrosine biosynthesis (prephenate dehydrogenase and arogenate dehydrogenase) were characterized in 90 species currently classified within the genera Pseudomonas, Xanthomonas, and Alcaligenes. Variation in cofactor specificity and regulatory properties of the dehydrogenase proteins allowed the separation of five groups. Taxa defined by enzymological patterning corresponded strikingly with the five ribosomal ribonucleic acid (rRNA) homology groups established via rRNA-deoxyribonucleic acid hybridization. rRNA homology groups I, IV, and V all lack activity for arogenate/nicotinamide adenine dinucleotide phosphate (NADP) dehydrogenase and separated on this criterion from groups II and III, which have the activity. Group II species possess arogenate dehydrogenase enzyme (reactive with either NAD or NADP) sensitive to feedback inhibition by tyrosine, thereby separating from group III species whose corresponding enzyme was totally insensitive to feedback inhibition. The presence of prephenate/NADP dehydrogenase in group IV defined its separation from groups I and V, which lack this enzyme activity. Group I species possess an arogenate/NAD dehydrogenase that was highly sensitive to inhibition by tyrosine and a prephenate/NAD dehydrogenase of relative insensitivity to tyrosine inhibition. The opposite pattern of sensitivity/insensitivity was seen in group V species. These dehydrogenase characterizations are highly reliable for the keying of a given species to one of the five rRNA homology groups. If necessary, other confirmatory assays can be included using other aromatic pathway enzymes. These results further document the validity and utility of the approach of comparative enzymology and allostery for classification of microorganisms.

Alcaligenes↗