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Applied and Environmental Microbiology Dec 1999The hemolytic activity and siderophore production of several strains of motile aeromonads were determined. The hemolytic activity of Aeromonas caviae and Aeromonas...
The hemolytic activity and siderophore production of several strains of motile aeromonads were determined. The hemolytic activity of Aeromonas caviae and Aeromonas eucrenophila was enhanced after trypsinization of the samples. The enhancement of hemolysis was observed in strains that carried an aerolysin-like gene, detected by a PCR procedure. Siderophore production was demonstrated in all but one strain of Aeromonas jandaei. No apparent relationship was observed between the presence of plasmid DNA and hemolysis or siderophore production.
Topics: Aeromonas; Animals; Bacterial Toxins; Esocidae; Hemolysin Proteins; Hemolysis; Oncorhynchus mykiss; Plasmids; Polymerase Chain Reaction; Pore Forming Cytotoxic Proteins; Rabbits; Sheep; Siderophores; Trout
PubMed: 10584028
DOI: 10.1128/AEM.65.12.5612-5614.1999 -
Journal of Clinical Microbiology Mar 1999Fifty-six isolates of four Aeromonas species, which have been documented as causative agents of human infections or isolated from human clinical specimens, were...
Fifty-six isolates of four Aeromonas species, which have been documented as causative agents of human infections or isolated from human clinical specimens, were subjected to antimicrobial susceptibility testing using a MicroScan WalkAway conventional (overnight incubation) gram-negative panel. The four species tested and the number of isolates of each were as follows: Aeromonas jandaei, 17; A. schubertii, 12; A. trota, 15; and A. veronii biotype veronii, 12. All isolates of A. trota were susceptible to all antimicrobial agents tested, except cefazolin (20% of isolates were resistant) and cefoxitin (13% of isolates were resistant). All isolates of A. schubertii and A. veronii biotype veronii, as well as 88% of A. jandaei isolates, were resistant to ampicillin. Resistance to ampicillin-sulbactam ranged from 25% of A. schubertii strains to 100% of A. veronii biotype veronii strains. Cefazolin resistance ranged from 17% of A. veronii biotype veronii isolates to 59% of A. jandaei isolates. Imipenem resistance was detected in 65% of A. jandaei strains and 67% of A. veronii biotype veronii strains. A. jandaei displayed resistance to piperacillin and ticarcillin in 53 and 71% of the isolates, respectively. A. veronii biotype veronii strains were 100% susceptible to piperacillin and 100% resistant to ticarcillin. These antibiogram data may be useful in establishing the identification of these four species when members of the genus Aeromonas are isolated from human clinical sources.
Topics: Aeromonas; Ampicillin Resistance; Anti-Bacterial Agents; Cefazolin; Cephalosporin Resistance; Gram-Negative Bacterial Infections; Humans; Microbial Sensitivity Tests; Serotyping
PubMed: 9986836
DOI: 10.1128/JCM.37.3.706-708.1999 -
Applied and Environmental Microbiology Feb 1998A low-specificity L-threonine aldolase (L-TA) gene from Pseudomonas sp. strain NCIMB 10558 was cloned and sequenced. The gene contains an open reading frame consisting...
A low-specificity L-threonine aldolase (L-TA) gene from Pseudomonas sp. strain NCIMB 10558 was cloned and sequenced. The gene contains an open reading frame consisting of 1,041 nucleotides corresponding to 346 amino acid residues. The gene was overexpressed in Escherichia coli cells, and the recombinant enzyme was purified and characterized. The enzyme, requiring pyridoxal 5'-phosphate as a coenzyme, is strictly L specific at the alpha position, whereas it cannot distinguish between threo and erythro forms at the beta position. In addition to threonine, the enzyme also acts on various other L-beta-hydroxy-alpha-amino acids, including L-beta-3,4-dihydroxyphenylserine, L-beta-3,4-methylenedioxyphenylserine, and L-beta-phenylserine. The predicted amino acid sequence displayed less than 20% identity with those of low-specificity L-TA from Saccharomyces cerevisiae, L-allo-threonine aldolase from Aeromonas jandaei, and four relevant hypothetical proteins from other microorganisms. However, lysine 207 of low-specificity L-TA from Pseudomonas sp. strain NCIMB 10558 was found to be completely conserved in these proteins. Site-directed mutagenesis experiments showed that substitution of Lys207 with Ala or Arg resulted in a significant loss of enzyme activity, with the corresponding disappearance of the absorption maximum at 420 nm. Thus, Lys207 of the L-TA probably functions as an essential catalytic residue, forming an internal Schiff base with the pyridoxal 5'-phosphate of the enzyme to catalyze the reversible aldol reaction.
Topics: Amino Acid Sequence; Binding Sites; Cloning, Molecular; Escherichia coli; Glycine Hydroxymethyltransferase; Hydrogen-Ion Concentration; Molecular Sequence Data; Molecular Weight; Pseudomonas; Substrate Specificity; Temperature
PubMed: 9464392
DOI: 10.1128/AEM.64.2.549-554.1998 -
Journal of Bacteriology Jun 1997We have isolated the gene encoding L-allo-threonine aldolase (L-allo-TA) from Aeromonas jandaei DK-39, a pyridoxal 5'-phosphate (PLP)-dependent enzyme that...
L-allo-threonine aldolase from Aeromonas jandaei DK-39: gene cloning, nucleotide sequencing, and identification of the pyridoxal 5'-phosphate-binding lysine residue by site-directed mutagenesis.
We have isolated the gene encoding L-allo-threonine aldolase (L-allo-TA) from Aeromonas jandaei DK-39, a pyridoxal 5'-phosphate (PLP)-dependent enzyme that stereospecifically catalyzes the interconversion of L-allo-threonine and glycine. The gene contains an open reading frame consisting of 1,014 nucleotides corresponding to 338 amino acid residues. The protein molecular weight was estimated to be 36,294, which is in good agreement with the subunit molecular weight of the enzyme determined by polyacrylamide gel electrophoresis. The enzyme was overexpressed in recombinant Escherichia coli cells and purified to homogeneity by one hydrophobic column chromatography step. The predicted amino acid sequence showed no significant similarity to those of the currently known PLP-dependent enzymes but displayed 40 and 41% identity with those of the hypothetical GLY1 protein of Saccharomyces cerevisiae and the GLY1-like protein of Caenorhabditis elegans, respectively. Accordingly, L-allo-TA might represent a new type of PLP-dependent enzyme. To determine the PLP-binding site of the enzyme, all of the three conserved lysine residues of L-allo-TA were replaced by alanine by site-directed mutagenesis. The purified mutant enzymes, K51A and K224A, showed properties similar to those of the wild type, while the mutant enzyme K199A was catalytically inactive, with corresponding disappearance of the absorption maximum at 420 nm. Thus, Lys199 of L-allo-TA probably functions as an essential catalytic residue forming an internal Schiff base with PLP of the enzyme to catalyze the reversible aldol reaction.
Topics: Aeromonas; Amino Acid Sequence; Base Sequence; Binding Sites; Cloning, Molecular; Genes, Bacterial; Glycine Hydroxymethyltransferase; Lysine; Molecular Sequence Data; Mutagenesis, Site-Directed; Pyridoxal Phosphate; Sequence Alignment
PubMed: 9171400
DOI: 10.1128/jb.179.11.3555-3560.1997 -
FEMS Immunology and Medical Microbiology Apr 1997An experimental study of five isolates of Aeromonas jandaei and 12 of A. trota was carried out to examine if they produced an enterotoxic substance, and if so, to...
An experimental study of five isolates of Aeromonas jandaei and 12 of A. trota was carried out to examine if they produced an enterotoxic substance, and if so, to characterise that factor and to see if it caused any mucosal damage. Only two of the A. trota strains caused fluid accumulation in the initial rabbit ileal loop (RIL) tests. The remaining strains did so only after one to five sequential passages through RILs and once they caused a secretory response they showed a gradual enhancement of fluid outpouring after each subsequent passage. Inocula of approximately 1 x 10(5) viable cells and 0.25 ml of culture filtrate caused fluid accumulations comparable to those of toxigenic V. cholerae 569B. The enterotoxic factors of both organisms were inactivated when held at 56 degrees C for 20 min or 65 degrees C for 10 min and showed biological activity over a wide range of pH. The only histopathological change observed in the ileal loop was depletion of mucus from the goblet cells. These data thus indicate that strains of A. jandaei and A. trota may produce a heat-labile and pH-stable diarrhoeagenic substance that causes little or no damage to the intestinal mucosa, like that of other known heat-labile enterotoxins.
Topics: Aeromonas; Animals; Enterotoxins; Hydrogen-Ion Concentration; Ileum; Rabbits
PubMed: 9143882
DOI: 10.1111/j.1574-695X.1997.tb01018.x -
Journal of Bacteriology Mar 1997Aeromonas jandaei AER 14 (formerly Aeromonas sobria AER 14) expresses three inducible beta-lactamases, AsbA1, OXA-12 (AsbB1), and AsbM1. Mutant strains that...
Aeromonas jandaei AER 14 (formerly Aeromonas sobria AER 14) expresses three inducible beta-lactamases, AsbA1, OXA-12 (AsbB1), and AsbM1. Mutant strains that constitutively overexpress all three enzyme simultaneously, suggesting that they share a common regulatory pathway, have been isolated. Detectable expression of the cloned genes of AsbA1 and OXA-12 in some Escherichia coli K-12 laboratory strains is achieved only in the presence of a blp mutation. These mutations map to the cre operon at 0 min, which encodes a classical two-component regulatory system of unknown function. Two regulatory elements from A. jandaei which permit high-level constitutive expression of OXA-12 in E. coli were cloned. Both loci encode proteins with characteristics of response regulator proteins of two-component regulatory systems. One of these loci, designated blrA, bestowed constitutive expression of all three beta-lactamases in A. jandaei AER 14 when present on a multicopy plasmid, confirming its role in the regulatory pathway of beta-lactamase production in this organism.
Topics: Aeromonas; Amino Acid Sequence; Bacterial Proteins; Base Sequence; Cloning, Molecular; Gene Expression Regulation, Bacterial; Molecular Sequence Data; Regulon; beta-Lactamases
PubMed: 9068648
DOI: 10.1128/jb.179.6.2006-2013.1997 -
Applied and Environmental Microbiology Nov 1995Aeromonas isolates were obtained from fish intestines, water, and sediments from an urban river and identified by the DNA-DNA microplate hybridization method. The...
Aeromonas isolates were obtained from fish intestines, water, and sediments from an urban river and identified by the DNA-DNA microplate hybridization method. The isolates were Aeromonas veronii (22%), Aeromonas caviae (18%), Aeromonas hydrophila (13%), Aeromonas sobria (8%), Aeromonas jandaei (7%), and other Aeromonas spp. (33%). Aeromonas species occurred at high densities with high incidences, regardless of season. The results strongly suggest that aeromonads are indigenous in fish intestines, water, and sediments of rivers and have the potential to be predominant in aquatic environments.
PubMed: 16535171
DOI: 10.1128/aem.61.11.4128-4130.1995 -
Applied and Environmental Microbiology Aug 1994Aeromonas isolates were obtained from the intestinal tracts of six species of cultured freshwater fish and identified on the basis of their genotypic and phenotypic...
Aeromonas isolates were obtained from the intestinal tracts of six species of cultured freshwater fish and identified on the basis of their genotypic and phenotypic characters. The microplate hybridization method could differentiate type strains of Aeromonas species and related bacteria. DNA-DNA hybridization analysis showed that 65 aeromonad isolates were 72 to 100% related with either Aeromonas caviae, Aeromonas hydrophila, Aeromonas jandaei, Aeromonas sobria, or Aeromonas veronii. As many as 48% of the genotypically identified A. caviae, A. hydrophila, and A. sobria isolates differed from the type strains of corresponding species in one to three phenotypic characters. These results strongly suggest that not all aeromonad isolates from freshwater fish could be identified correctly on the basis of only the phenotypic characters, indicating the usefulness of the microplate hybridization method for the identification of aeromonads.
PubMed: 16349363
DOI: 10.1128/aem.60.8.3036-3038.1994 -
Journal of Clinical Microbiology Mar 1991Exudate removed from an infection that developed below the left eye of a 10-year-old male following a previously inflicted wound after aquatic exposure was cultured and...
Exudate removed from an infection that developed below the left eye of a 10-year-old male following a previously inflicted wound after aquatic exposure was cultured and revealed two different Aeromonas spp. Further characterization showed that one strain was phenotypically identical to Aeromonas veronii, while the other strain was confirmed by DNA hybridization analysis to be Aeromonas jandaei sp. nov. This is the first report of these more recently described aeromonads, thus far rarely reported from clinical disease, occurring simultaneously in a human infection.
Topics: Aeromonas; Bacterial Infections; Child; DNA, Bacterial; Drug Resistance, Microbial; Fresh Water; Humans; Male; Nucleic Acid Hybridization; Species Specificity; Water Microbiology; Wound Infection
PubMed: 2037674
DOI: 10.1128/jcm.29.3.565-569.1991 -
Journal of Clinical Microbiology Mar 1991A large numerical taxonomy study conducted in 1988 of 165 mostly clinical Aeromonas strains from diverse geographic sources produced a cluster (S = 84%, SSM) of four...
A large numerical taxonomy study conducted in 1988 of 165 mostly clinical Aeromonas strains from diverse geographic sources produced a cluster (S = 84%, SSM) of four sucrose-negative strains that included the DNA definition strain for DNA group 9 A. sobria (CDC 0787-80). These four strains, together with five additional strains received in 1989, were subjected to DNA-DNA hybridization (hydroxyapatite, 32P, 60 and 75 degrees C), and all eight strains were closely related to the ninth labeled DNA group 9 definition strain CDC 0787-80 (73 to 86% relatedness at 60 degrees C and 68 to 80% relatedness at 75 degrees C; percent divergence, 2.0 to 3.5). Type strains and DNA definition strains for all other established Aeromonas species were only 35 to 72% related (60 degrees C) to CDC 0787-80. We propose the name Aeromonas jandaei for this highly related group of nine strains, formerly known as DNA group 9 A. sobria. The type strain was designated ATCC 49568 (CDC 0787-80). The nine strains were examined at 36 degrees C and were found to be resistant to 0/129 (vibriostatic agent) and uniformly positive for oxidase, gas production from glucose, indole, lysine decarboxylase, arginine dihydrolase, o-nitrophenyl-beta-D-galactopyranoside, motility (25 degrees C), nitrate reduction, citrate utilization, hemolysis on sheep blood agar, and growth in Trypticase soy broth with no added NaCl. They all fermented D-glucose, D-mannitol, and mannose but did not ferment sucrose, cellobiose, L-arabinose, inositol, salicin, or D-sorbitol. They were uniformly negative for esculin and urea hydrolysis, elastase production, ornithine decarboxylation, and the string test. The antibiogram of A. jandaei resembled that of other aeromonads (resistance to ampicillin and cephalothin), but it differed from most other aeromonads because of resistance to single dilution of colistin and differed from clinical A. veronii biogroup sorbria (formerly A. sobria) by its nearly uniform resistance to cephalothin. The esculin-, sucrose-, and cellobiose-negative and colistin-resistant profile distinguished A. jandaei from other Aeromonas species. These A. jandaei strains were isolated from blood (two strains), wounds (two strains), diarrheal stools (four strains), and a prawn (one strain). The blood and wound isolates, in particular, suggest that there is a possible clinical significance for this species and justify identification of and further research on this group of motile aeromonads.
Topics: Aeromonas; Aged; Bacterial Infections; Child; DNA, Bacterial; Drug Resistance, Microbial; Humans; Male; Nucleic Acid Hybridization; Phenotype; Sepsis; Species Specificity; Sucrose
PubMed: 2037673
DOI: 10.1128/jcm.29.3.560-564.1991