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Genomic Characterization of Aeromonas dhakensis Isolated From a Fatal Dolphin Case.

Aeromonas dhakensis has emerged as a significant pathogen affecting both aquatic animals and humans; however, genomic data for isolates from marine mammals remain scarce. In this study, we characterised the genome of A. dhakensis strain KDL-001, isolated from a fatal dolphin case, using whole-genome sequencing and comparative genomics. Taxonomic analyses, including MLST and average nucleotide identity (ANI), confirmed the isolate as A. dhakensis. Core-genome phylogeny further revealed that KDL-001 is closely related to strains derived from fish and aquatic environments. Notably, in silico screening of virulence-associated genes showed that the virulence-associated gene profile of the dolphin isolate was broadly comparable to those of other A. dhakensis strains, with no isolate-specific virulence-associated genes being identified within the limits of this analysis. These findings demonstrate that the dolphin-derived isolate is genomically comparable to previously described A. dhakensis strains and possesses conserved virulence-associated genes commonly found within the species.

Animals↗

Evaluation of two miniaturized systems, MicroScan W/A and BBL Crystal E/NF, for identification of clinical isolates of Aeromonas spp.

Fifty-two clinical strains and 22 type and reference Aeromonas strains, previously genetically characterized by 16S rRNA gene restriction fragment length polymorphism, were identified in parallel with the MicroScan Walk/Away and BBL Crystal Enteric/Nonfermenter systems. The former identified only 14.8% of the isolates correctly, and the latter identified only 20.3% correctly, which indicates that neither of these systems is useful for this purpose.

Aeromonas↗

Comparison of two assays for the detection of haemolysins of Aeromonas species.

The haemolysins produced by Aeromonas species were detected and compared by two assay methods--a modified blood agar plate assay and the rabbit erythrocyte haemolysin method. Both assays showed a high level of agreement (86%). The titres of the rabbit erythrocyte haemolysin assay correlated with the haemolytic zone diameter of the ox blood agar assay. In addition the agar haemolysin assay had simple media requirements, was easy to perform and results were well defined.

Aeromonas↗

[The pattern of the sensitivity of Aeromonas spp. that produce extraintestinal infections].

We studied the sensibility to several antimicrobics of aeromonas strains isolated in extraintestinal infections during a period of five years, in order to establish which could be the best empirical therapy in these processes. Twenty-two strains of A. hydrophila were evaluated, was isolated (8 from hemocultures, 6 from wound exudate, 2 from abscess aspirate, 2 from peritoneal fluid and 1 from urine, pleural fluid, bile and catheter). The identification was made using the GNI card System (Vitek System) and other tests. The sensibility was assessed using the GNS-BH and GNS-BI card System (Vitek), which allows to know the clinical category and CMI value in microgram/ml. All the strains presented some resistance to antibiotics. 77.8% of the strains were resistant to more than one antibiotic and 64.9%, to more than two antibiotics. We detected 21.7% strains resistant to 4 drugs. The most frequent resistance association was ampicillin, cephoxitine, cephazoline, cephalotine, with or without amoxicillin and clavulanic acid. All the strains were sensible to cephamandol, cephuroxime, cephtriaxone, cephtazidime, cephotaxime, thicarciline, aztreonam, azlociline, mezlociline, piperaciline, gentamycin, amikacine, chloranphenicol and ciprofloxacine. All the strains were resistant to ampicillin; 60.9% of the strains were sensible to the association amoxicillin and clavulonic acid. In conclusion, the treatment of extraintestinal infections does not differs basically from the treatment of the infections caused by Gram-negative bacillus. There are a broad range of effective antibiotics: aminoglycosides (gentamycin, tobramycin, amikacine), piperacyline, cephalosporines (cephuroxime, cephotaxime, cephtazidime and cephtriaxone), chloranphenicol, penemas, monobactanes of fluorquinolone.

Aeromonas↗

[Aeromonas spp. and Plesiomonas shigelloides in bivalves, mud, and water from the Gulf of Nicoya, Costa Rica].

Bivalves, mud, and surface water were collected at three different sites of the Gulf of Nicoya, Costa Rica, in search of Aeromonas spp. and Plesiomonas shigelloides. For their isolation, these bacteria were enriched in alkaline peptone water and streaked on MacConkey agar and on brilliant green bile inositol agar. This was followed by the biochemical tests necessary for their identification. Thirty-five strains of A. hydrophila, 58 of A. caviae, 43 of A. sobria, and 7 of P. shigelloides were isolated. None of these predominated nor was there any indication of a seasonal distribution along the 15 month's duration of the study. Seven strains of A. hydrophila and two of A. sobria showed the biochemical characteristics associated with toxin production (positive Voges-Proskauer and lysine decarboxylase tests). These species are widely distributed in the gulf and there is risk of contracting an infection while bathing or when eating raw bivalves from this area.

Aeromonas↗

Cholera-like diarrhea in Canada. Report of a case associated with enterotoxigenic Escherichia coli and a toxin-producing Aeromonas hydrophila.

A 67-year-old Indian patient was admitted with an acute cholera-like illness. Toxigenic Escherichia coli producing both heat-stable and heat-labile enterotoxins grew from cultures of feces. In addition, an Aeromonas hydrophila producing a cytotoxic toxin was also isolated from this patient's feces. The unusual severity of this patient's illness may have resulted from coinfection with these two toxigenic organisms, although any role of the toxin produced by a hydrophila is speculative.

Aeromonas↗

Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by high-cell-density cultivation of Aeromonas hydrophila.

The newly screened Aeromonas hydrophila produces copolymer consisting of 3-hydroxybutyrate (3HB) and 3-hydroxyhexanoate (3HHx). The characteristics of cell growth and polymer accumulation were examined using various carbon sources. P(3HB-co-3HHx) was produced from lauric acid and oleic acid only. P(3HB-co-3HHx) content can be increased by limitation of phosphorus. A maximal P(3HB-co-3HHx) content of 28.8 wt% could be obtained in flask culture. By applying the optimally designed nutrient feeding strategy, cell dry weight, P(3HB-co-3HHx) content, and 3HHx fraction obtained over the course of 43 h were 95.7 g/L, 45.2 wt%, and 17 mol%, respectively, resulting in a productivity of 1.01 g polyhydroxyalkanoate (PHA)/L. h.

3-Hydroxybutyric Acid↗

Antibacterial activity of chitosan against Aeromonas hydrophila.

The effect of chitosan on growth and production of haemolysin by Aeromonas hydrophila was investigated as well as the effects of temperature, pH, salts and irradiation on the antibacterial activity of chitosan. It was found that chitosan affected growth and haemolysin production of A. hydrophila in varying degrees compared to the control. Growth and haemolysin production were clearly suppressed at 0.04% of chitosan. Suppression was more effective at pH 6.0 than at pH 7.0. The bactericidal effects of chitosan increased with increasing temperature and decreasing pH. Divalent cations at concentrations of 10 and 25 mM reduced the antibacterial activity of chitosan, in the order of Ba2+ > Ca2+ > Mg2+. Sodium ions at concentrations of 10 and 25 mM also reduced chitosan's activity. Irradiation of chitosan at 150 kGy under dry condition was effective in slightly increasing its activity.

Aeromonas hydrophila↗

Acute arthritis caused by Aeromonas hydrophola: clinical and therapeutic aspects.

Two cases of Aeromonas hydrophila arthritis are reported and a third case present in the literature is reviewed. All three cases occurred in patients with myelogenous leukemia; the knee joint was involved in 2 patients and the metacarpal-phalangeal joint in the third. In the 2 patients presented here, synovial fluid antibiotic levels were measured and were adequate; but A hydrophila was repeatedly cultured from the knee joints, a fact suggesting that other factors, such as normal functioning leukocytes and specific opsonizing antibody against the organism, are also necessary to kill A hydrophila.

Acute Disease↗

Parapharyngeal soft-tissue infection with Aeromonas hydrophila.

Parapharyngeal soft tissue infections may be rapidly progressive and life-threatening. Prompt institution of appropriate antimicrobial therapy is of paramount importance. This report highlights the potential virulence of Aeromonas hydrophila in infection of the head and neck and the need to consider this organism in selected patients.

Adult↗

Influence of temperature, salinity and pH on the growth of environmental Aeromonas and Vibrio species isolated from Mai Po and the Inner Deep Bay Nature Reserve Ramsar Site of Hong Kong.

Four environmental bacterial isolates including Aeromonas hydrophila MP-3, A. salmonicida MP-4, Vibrio vulnificus MP-2 and V. cholerae MP-1 isolated from sediment and water of Mai Po Nature Reserve of Hong Kong were examined for their responses to temperature, pH and salinity under laboratory conditions in this study. V. cholerae MP-1 was found to resist vibriostatic agent O/129 at concentration of 10 microg/ml. In addition, bacterial growth under test conditions was measured and the results were fitted into the Gompertz model to obtain important parameters related to bacterial growth, lag time (lambda), specific growth rate (mu(m)), and maximum biomass (A) for comparison. V. cholerae MP-1 did not show any apparent growth at 15 degrees C, but was adapted to a much wider environmental pH from 5.2 to 9.2 for growth while V. vulnificus MP-2 was more sensitive to pH changes yielding the highest biomass at pH 6.2. A. salmonicida MP-4 was surprisingly tolerant to salinity as high as 60.0 per thousand NaCl and grew almost equally well as under conditions of other treatments. All four bacterial isolates showed a wide spectrum of plasticity to the environmental conditions and they pose a potential threat to public health and animal health.

Aeromonas↗

Mechanistic studies of two amino acid racemases of broad substrate specificity from Pseudomonas striata and Aeromonas caviae.

The conversion of L-[alpha-2H]alanine in H2O and unlabeled L-alanine in 2H2O into D-alanine, under nearly irreversible conditions, with the amino acid racemase from Pseudomonas striata showed significant internal transfer of the alpha-hydrogen. This result has been interpreted as being indicative of a single base mechanism for the racemization. The relative rates of deuterium incorporation into unlabeled D- and L-methionine by the two amino acid racemases of broad substrate specificity from P. striata and Aeromonas caviae, were measured in 2H2O. The results showed a markedly different pattern, dependent upon the configuration of the initial substrate; with D-methionine as substrate deuterium is incorporated into both enantiomers at approximately the same rate, but with L-methionine as substrate deuterium is incorporated considerably faster into the D than the L enantiomer. These results argue against a single base mechanism of racemization for these enzymes and are best rationalized in terms of a double base model where only one of the bases undergoes proton (deuterium) exchange with the solvent while the amino acid is enzyme-bound. The interpretation of the earlier experiment needs to be considered in light of these results.

Aeromonas↗

Factors influencing beta-galactosidase activity of Aeromonas caviae.

Aeromonas caviae, often reported to be associated with diarrhoeal patients, elaborates several virulence factors as well as catabolic enzymes such as xylanase and beta-galactosidase. Studies on the kinetics of growth of A. caviae and synthesis of beta-galactosidase suggested that the activity was cell associated and reached a peak during the late logarithmic phase of growth. The optimum pH for beta-galactosidase activity was 7.0 and required Ca2+ and glutathione for enhancement of its activity; IPTG also slightly improved the activity. Aerobic cultivation of A. caviae in LB containing glucose, fructose, maltose and sucrose completely inhibited the activity possibly due to acetic acid production. Addition of 100 mM cAMP to the media containing glucose (0.25%, w/v) restored the relative activity by 8.8%; however, the final pH of the media remained acidic. Aerobic growth of A. caviae with other carbon sources did not affect beta-galactosidase activity, probably as there was no acid production and thereby the final pH of the media unaltered. Arabinose, xylose and galactose induced the A. caviae beta-galactosidase activity by several folds and lactose moderately enhanced its activity.

Aeromonas↗

Metabolic engineering for the production of copolyesters consisting of 3-hydroxybutyrate and 3-hydroxyhexanoate by Aeromonas hydrophila.

Aeromonas hydrophila 4AK4 was able to synthesize copolyesters consisting of 3-hydroxybutyrate (3HB) and about 15 mol-% 3-hydroxyhexanoate (3HHx) (PHBHHx) when grown in long chain fatty acids such as dodecanoate regardless of growth conditions. To regulate the unit fraction in PHBHHx, phbA and phbB genes encoding beta-ketothiolase and acetoacetyl-CoA reductase in Ralstonia eutropha, were introduced into A. hydrophila 4AK4. When gluconate was used as cosubstrate of dodecanoate, the recombinant produced PHBHHx containing 3-12 mol-% 3HHx, depending on the gluconate concentration in media. Vitreoscilla hemoglobin gene, vgb, was also introduced into the above recombinant, resulting in improved PHBHHx content from 38 to 48 wt.-% in shake flask study. Fermentor studies also showed that increased gluconate concentration in medium containing dodecanoate promoted the recombinant strain harboring phbA and phbB genes to incorporate more 3HB unit into PHBHHx, resulting in reduced 3HHx fraction. Recombinant A. hydrophila harboring phbA, phbB and vgb genes demonstrated better PHBHHx productivity and higher conversion efficiency from dodecanoate to PHBHHx than those of the recombinant without vgb in fermentation study. Combined with the robust growth property and simple growth requirement, A. hydrophila 4AK4 appeared to be a useful organism for metabolic engineering.

3-Hydroxybutyric Acid↗

Differential proteomic analysis of Aeromonas salmonicida outer membrane proteins in response to low iron and in vivo growth conditions.

Aeromonas salmonicida subsp. salmonicida is the etiological agent of furunculosis, a serious infectious disease of salmonids. Bacterial phenotypes are known to change in vivo compared to the in vitro state. Proteomic analysis of in vivo phenotypes is usually not possible due to insufficient biomass. Using an in vivo growth chamber model, the pathogenic fish bacterium A. salmonicida was cultured in pure culture in vivo in its host, the Atlantic salmon, to obtain sufficient biomass to allow proteomic analysis. Growth of A. salmonicida under in vitro iron-restricted conditions resulted in the expression of outer membrane proteins of 73, 76 and 85 kDa, which were not present when grown under in vitro iron-replete conditions. Mass spectrometry analysis identified the 73 kDa protein as a colicin receptor, the 76 kDa protein as an outer membrane heme receptor, and the 85 kDa protein as a ferric siderophore receptor. When cultured in vivo, A. salmonicida up-regulated the identical 73, 76 and 85 kDa proteins. The results of this study also suggest, at least with respect to the outer membrane proteins, that the in vitro iron-restricted growth model largely reproduces the results obtained from growth of A. salmonicida within the peritoneal cavity of salmon.

Aeromonas↗