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We describe a case of Aeromonas hydrophila endocarditis in a 66-year-old man with myelodysplastic syndrome and non-A, non-B hepatitis, The infection resolved with antibiotic therapy, but the patient succumbed to complications of his underlying illness. This is the second case of Aeromonas endocarditis reported in the world literature.
A correlation between the genospecies (DNA-DNA hybridization group) and the type of siderophore produced by 118 isolates of the genus Aeromonas was established. Organisms in hybridization groups 1 through 5 (including 5A, 5B, and 5AB) and group 12 predominantly produced the siderophore amonabactin, while an enterobactinlike siderophore was prevalent in groups 8/10 and 9. The siderophore produced by strains in group 6 may be an as-yet-unidentified nonphenolate, nonhydroxamate compound, and group 7 isolates synthesized no siderophores. Determination of the indigeneous siderophore (or the absence of one) produced by an isolate of the genus Aeromonas may assist in identification of the organism's genetic species and may suggest the presence of certain virulence properties.
Aeromonas caviae has recently been recognized as an important enteropathogen and its hemolysin is purported to be one of the virulence factors, In this study, a total of 80 clinical isolates of Aeromonas spp. were investigated by PCR with synthetic oligonucleotides targeting a cloned hemolysin-encoding sequence from an A. caviae isolate of clinical origin. Of the 35 clinical A. caviae isolates tested, only 6 contained the target sequence.
We characterized a collection of 268 Aeromonas isolates from diverse sources (clinical, animal, and environmental sources) for their species and serogroup designations. Overall, 97% of these strains could be identified to the genomospecies level by using an expanded battery of biochemical tests. Members of the Aeromonas hydrophila complex (A. hydrophila, HG2, and A. salmonicida), a group that has previously been difficult to separate biochemically, could easily be distinguished from one another by using a number of recently described phenotypic properties which included utilization of DL-lactate and urocanic acid. Differences in species distributions on the basis of the source of isolation were noted. Serogroup analysis of these 268 isolates plus a number of reference cultures indicated that (i) each genomospecies is serologically heterogeneous and individual serogroups can be found in more than one species, (ii) most type or reference strains for each hybridization group are not serologically representative of the genomospecies at large, (iii) serogroups O:11, O:34, and O:16 predominate clinically (48%), supporting previous studies indicating their importance in human infections, and (iv) most A. trota strains do not express the O139 antigen of Vibrio cholerae. The collective results suggest that both species and serogroup designations are important factors in establishing which isolates can cause human infections when they are acquired from nonclinical sources (foods, animals, and the environment).
Aeromonas isolates from patients with diarrhea in Bangladesh (n = 69), from healthy controls (n = 11), and from surface water (n = 40) were analyzed with respect to their hybridization groups (HGs) by the aid of fatty acid methyl ester (FAME) characterization and DNA fingerprinting by AFLP, biochemical phenotypes (Phe-nePlate [PhP] types), and the production of hemolysin and cytotoxin. The aim of the investigation was to find out whether certain strains carrying virulence factors predominated among patient isolates. According to FAME and/or AFLP analysis, most human isolates were allocated to DNA HGs 4 (Aeromonas caviae) and 1 (A. hydrophila). Most environmental strains were allocated to HG8 (A. veronii biogroup sobria) and HG4 (A. caviae), and only one was of HG1. According to PhP typing, the diversity among patient isolates was lower than that among other strains, and two dominating PhP types (types BD-1 and BD-2) were identified in 29 and 30% of the patient isolates, respectively. PhP type BD-1 was also common among the environmental isolates, whereas PhP type BD-2 was only identified in two of the other isolates. Twenty-five of 26 isolates belonging to HG1 were of the same PhP type (BD-2), whereas isolates of other common HGs were more diverse according to their PhP types. Hemolytic and cytotoxin-producing strains occurred more frequently among the environmental isolates than among patient isolates. However, the hemolytic and cytotoxic activities among human isolates was strongly correlated to the HG1/BD-2 type, which, in addition, showed high cytotoxin titers (median values, 1/512 compared to 1/128 for cytotoxin-positive isolates belonging to other types). Thus, the HG1/BD-2 type may represent a pathogenic A. hydrophila type that is able to produce diarrhea in humans.
Identification of Aeromonas species, emergent pathogens for humans, has long been controversial due to their phenotypic and genomic heterogeneities. Computer analysis of the published 16S rRNA gene sequences revealed that restriction fragment length polymorphism of the PCR-amplified 16S rRNA gene is a good and rapid way of assessing the identities of all known species of Aeromonas. The method was evaluated with the reference strains of all species (or DNA homology groups) and 76 clinical isolates of diverse origin. Most results from the two approaches were in agreement, but some discrepancies were discerned. Advantages over previous phenotypic and genetic methods are discussed.
Aeromonas popoffii is a recently described species isolated mainly from freshwater. An isolate of Aeromonas popoffii was found to be responsible for a urinary tract infection in a 13-year-old boy suffering from spina bifida with enterocystoplasty. This is the first reported case of human infection attributed to this species.
Eleven isolates of Aeromonas species from human stool cultures were found to produce enterotoxin activity as determined by assay of culture filtrates in rabbit intestinal loops and rabbit skin and on adrenal Y1 cells. Hemolysin(s) and a cytotoxic protein were found to interfere in all three assay systems but could be inactivated upon heating at 56 degrees C or by specific antihemolysin. Biotyping of each isolate was performed with a conventional test system and with API 50E and APIZYM kit systems (Analytab, Inc.). No single test of the more than 70 biochemical reactions investigated was found to correlate with enterotoxigenicity in the strains of Aeromonas hydrophila examined. All strains were found to belong to ideal phenotypes of A. hydrophila, but each strain possessed its own biochemical profile.
Over a four and a half month period, 1004 unselected routine faecal specimens from 815 patients were cultured for Aeromonas hydrophila. Forty-two specimens (4.2%) representing 38 patients were culture-positive. The study specimens also yielded Salmonella on 116 occasions, Shigella on seven, Campylobacter species on six and other bacterial pathogens on 17 occasions, respectively. Seven specimens had A hydrophila together with another bacterial pathogen. In only 19 of 38 patients (50%) was A hydrophila possibly associated with gastrointestinal symptoms. All the Aeromonas isolates were resistant to ampicillin but sensitive to gentamicin, piperacillin, mecillinam, chloramphenicol, ceftazidime and colistin.
Motile Aeromonas isolated from fish were studied for their virulence in fish in relation to some surface characteristics. The results showed that only the most virulent strains of A. hydrophila used in this study shared a common O antigen, did not agglutinate in acriflavine, settled down after boiling, and were resistant to the bactericidal action of fresh normal mammalian serum. The least virulent strains could not be grouped into this O antigenic group, they did not settle after boiling, and were sensitive to the bactericidal effect of serum. It is suggested that agglutination in acriflavine, stability after boiling, and sensitivity to normal fresh serum could be used for screening the Aeromonas isolates for virulence in fish.
Eleven ornithine-positive strains of Aeromonas (9 A. veronii and 2 provisionally classified as Aeromonas species ornithine positive) were tested for ability to cause fluid accumulation in the rabbit ileal loop. All eight beta-hemolytic strains caused fluid accumulation. Gel diffusion analysis revealed that the A. veronii beta-hemolysin was serologically related to the A. hydrophila beta-hemolysin, a known enterotoxic molecule. The biological activity of the A. veronii hemolysin was neutralized by antiserum to A. hydrophila hemolysin. One of three strains that were not beta-hemolytic caused fluid accumulation but only when the ileal loops were inoculated with live cultures. These results suggest that A. veronii is a potential enteropathogen that can cause diarrhea by means of a cell-freed enterotoxin (beta-hemolysin) or by a second mechanism that requires the presence of whole cells.
When grown in vivo in the peritoneal cavity of rainbow trout, Aeromonas salmonicida produces a clearly defined capsule with virulence-related functions. Aeromonas salmonicida grown in vitro in a glucose-rich medium (GRM) has also been reported to reproduce capsular material. Because in vitro mimicry of in vivo induced traits is highly desirable in vaccine design, the extent to which growth in GRM mimicked in vivo growth was examined. Antibodies specific to in vivo grown cells partially labeled the surface of GRM-grown cells, as well as two distinct proteins (81,700 and 41,000 Mr) in immunoblots of mutants with S-layer or lipopolysaccharide defects. GRM-grown strains showed an increased sensitivity to trout serum in contradistinction to the complete serum resistance of in vivo grown cells; as well, GRM-grown cells were more adherent to trout macrophages. Thus in spite of possessing some surface antigens normally expressed in vivo, cells grown on solid GRM did not possess all functional properties of in vivo grown cells.
Aeromonas hydrophila and Aeromonas salmonicida were shown to release "blebs" or fragments averaging 20-30 nm in diameter. The protein composition of the fragments was very similar to that of the corresponding outer membrane by sodium dodecyl sulphate--polyacrylamide gel electrophoresis and the fragments were shown to contain phospholipid and lipopolysaccharide. The results therefore indicate that the blebs are derived from the outer membrane of these organisms. Fragments isolated directly by differential ultracentrifugation were indistinguishable from fragments isolated by salt fractionation and gel filtration in chemical composition, protein composition, and density. However fragments isolated directly contained much less glycerophospholipid-cholesterol acyltransferase than those isolated by salt fractionation. The potential usefulness of membrane fragments to the bacteria and as a tool in the study of outer membrane structure and function is discussed.
Existing biochemical methods cannot distinguish among some species of Aeromonads, while genetic methods are labor intensive. In this study, primers were developed to three genes of Aeromonas: lipase, elastase, and DNA gyraseB. In addition, six previously described primer sets, five corresponding to species-specific signature regions of the 16S rRNA gene from A. veronii, A. popoffii, A. caviae, A. jandaei, and A. schubertii, respectively, and one corresponding to A. hydrophila specific lipase (hydrolipase), were chosen. The primer sets were combined in a series of multiplex-PCR (mPCR) assays against 38 previously characterized strains. Following PCR, each species was distinguished by the production of a unique combination of amplicons. When the assays were tested using 63 drinking water isolates, there was complete agreement in the species identification (ID) for 59 isolates, with ID established by biochemical assays. Sequencing the gyrB and the 16S rRNA gene from the remaining four strains established that the ID obtained by mPCR was correct for three strains. For only one strain, no consensus ID could be obtained. A rapid and reliable method for identification of different Aeromonas species is proposed that does not require restriction enzyme digestions, thus simplifying and speeding up the process.
BACKGROUND: Aeromonas salmonicida has been isolated from numerous fish species and shows wide variation in virulence and pathogenicity. As part of a larger research program to identify virulence genes and candidates for vaccine development, a DNA microarray was constructed using a subset of 2024 genes from the draft genome sequence of A. salmonicida subsp. salmonicida strain A449. The microarray included genes encoding known virulence-associated factors in A. salmonicida and homologs of virulence genes of other pathogens. We used microarray-based comparative genomic hybridizations (M-CGH) to compare selected A. salmonicida sub-species and other Aeromonas species from different hosts and geographic locations. RESULTS: Results showed variable carriage of virulence-associated genes and generally increased variation in gene content across sub-species and species boundaries. The greatest variation was observed among genes associated with plasmids and transposons. There was little correlation between geographic region and degree of variation for all isolates tested. CONCLUSION: We have used the M-CGH technique to identify subsets of conserved genes from amongst this set of A. salmonicida virulence genes for further investigation as potential vaccine candidates. Unlike other bacterial characterization methods that use a small number of gene or DNA-based functions, M-CGH examines thousands of genes and/or whole genomes and thus is a more comprehensive analytical tool for veterinary or even human health research.
Five restriction endonucleases (ENases) and one ENase were found in a screen of 196 strains of Plesiomonas shigelloides and 147 strains of Aeromonas species. Plesiomonas and Aeromonas species are classified as Vibrionaceae, identified as food-poisoning bacteria, are closely genetically related to each other, and their ENases producing abilities have not bee reported. ENases were detected at relatively low frequencies in these species as compared to those in other species, such as Salmonella species and Vibrio parahaemolyticus. All Enases were shown to be isoschizomers of already known ENases. One of the Plesiomonas ENases, designated PshBI, recognizing the sequence 5'-AT/TAAT-3' should be useful, since PshBI ENase is produced at a high yield of 7000 units/g of wet cells. The specificities of other ENases are also described in this paper.
A flake-chitin degrading marine bacterium was isolated and identified as Aeromonas hydrophila. This strain secreted five chitinases and an beta-N-acetylglucosaminidase. The main chitinase (Chi-A) was purified and characterized. The optimum pH of Chi-A was 5-8, and the activity was inhibited by Hg2+ and Fe3+. Chi-A was different from chitinases of other Aeromonas species with respect to molecular weight (62,000) and insensitivity to monoiodoacetate. The amino-terminal amino acid sequence showed extensive similarity with chitinases from Gram-negative bacteria.
The genus Aeromonas comprises known virulent and avirulent isolates and has been implicated in waterborne disease. A common infection model of human gastroenteritis associated with A. hydrophila uses neonatal mice. The goal of this research was to evaluate whether a murine small intestinal cell line could provide comparable results to the gene expression changes in the neonatal mouse model. Changes in mRNA expression in host cell cultures and intestinal tissues were measured after exposure to virulent Aeromonas hydrophila strains. A. hydrophila caused the up-regulation of more than 200 genes in neonates and over 50 genes in cell culture. Twenty-six genes were found to be in common between the two models, of which the majority are associated with the innate immune response.