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In vitro susceptibility of Aeromonas caviae, Aeromonas hydrophila and Aeromonas sobria to fifteen antibacterial agents.

In vitro testing of the activity of 15 antibacterial agents against 522 clinical isolates of Aeromonas species demonstrated some species-associated trends. Amoxicillin plus clavulanic acid was effective against approximately 45% of Aeromonas caviae and Aeromonas hydrophila, but all Aeromonas sobria isolates were resistant. Aztreonam, piperacillin and mezlocillin were highly active against all the strains of Aeromonas tested. Ticarcillin was equally effective against Aeromonas caviae and Aeromonas hydrophila, but more than 50% of Aeromonas sobria isolates were resistant. The latter species was more susceptible to cephalosporins than Aeromonas hydrophila and Aeromonas caviae. Chloramphenicol, tetracycline and trimethoprim-sulfamethoxazole were extremely active against all three Aeromonas species, likewise ofloxacin and ciprofloxacin. Aztreonam, third-generation cephalosporins, chloramphenicol and the quinolones can thus be considered for therapy of infections when Aeromonas is implicated.

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↗

Proteolytic activity of Aeromonas caviae.

Aeromonas strains produce a variety of virulence factors including proteases. Studies on the kinetics of growth of Aeromonas caviae NRRL B-966 and its proteases suggest that the proteolytic activities are produced throughout the growth phase, with peak level occurring at stationary phase. A. caviae synthesize both intracellular and extracellular proteases with the latter account for major portion of the total activity. Optimum pH for the A. caviae proteolytic activity is at 7.0. A. caviae produces a thermoresistant protease, whose activity is dependent on Mg++ and Ca++ ions. Inhibition of proteolytic activity by phenyl methyl sulfonyl fluoride suggest the presence of a serine protease in A. caviae. Nitrogenous compounds enhance the proteolytic activity while carbohydrates tested in this study inhibit the activity.

Aeromonas↗

Characterisation of haemolytic activity from Aeromonas caviae.

Aeromonas caviae, an enteropathogen associated with gastroenteritis, displays several virulence characteristics. Studies on the kinetics of growth of A. caviae and expression of beta-haemolytic toxin revealed that A. caviae produced maximum haemolytic activity extracellularly during the stationary phase. Preliminary studies on the properties of A. caviae haemolysin suggested that divalent cations (Mg2+ and Ca2+) and thiol compounds, dithiothreitol and mercaptoethanol enhanced the haemolytic activity. Addition of L-cysteine, glutathione and EDTA reduced the haemolytic activity. The iron chelator, 2-2' bipyridyl, significantly inhibited the growth of A. caviae possibly by iron limitation, with parallel enhancement of haemolysin production compared to A. caviae grown in excess of iron. These results suggest that A. caviae produces only beta-haemolysin, which resembles the haemolysins reported for several other bacteria and the activity might be regulated by environmental factors especially iron.

2,2'-Dipyridyl↗

Rapid and sensitive method for the detection of Aeromonas caviae and Aeromonas trota by polymerase chain reaction.

A 16S rDNA-based polymerase chain reaction (PCR) method was developed for the detection of Aeromonas caviae and Aeromonas trota. These two species were identified from other Aeromonas spp. and closely related species by primers set (AER1 and AER2). The amplified product was 316 bp. The identity of the amplified product was confirmed by DNA-DNA hybridization. Two sets of primers (AER8 and AER9) were used for specific identification of Aer. caviae. Amplifying the 260 bp fragment of 16S rRNA gene region and digesting it with AluI restriction enzyme, yielded 180- and 80-bp fragments. For PCR assay, template DNA was released by mixing equal volumes of homogenized seeded crab meat with Aer. caviae and Chelex 100 (6%) incubated for 10 min at 56 degrees C followed by addition of an equal volume of 0.1% Triton-X-100 and boiled for 10 min. The detection limit was between 50 and 100 cells g-1 of crab meat. This method is very rapid and obviates the need for DNA isolation from complex food matrices and is specific for detecting two Aeromonas species.

Aeromonas↗

Identification of a 43-kDa outer-membrane protein as an adhesin in Aeromonas caviae.

Aeromonas spp. are associated with intestinal and extra-intestinal infections. However, the virulence factors of A. caviae remain, for the most part, poorly known. This study examined the interactions involved in the adherence of A. caviae isolates Ae56, Ae391 and Ae398 to HEp-2 cells. All strains expressed high levels of aggregative adherence. Maximum adhesion occurred with bacteria grown at 22 degrees C, but transmission electron microscopy did not reveal the presence of fimbrial structures on the bacterial cell surface. Outer-membrane proteins (OMPs) extracted from isolate Ae398, grown at 22 degrees C and 37 degrees C, showed similar SDS-PAGE protein profiles. Most proteins were < 60 kDa. A major 43-kDa protein was seen only in the boiled OMP extract. The biotinylated 43-kDa protein bound specifically to HEp-2 cells. Microbeads coated with the 43-kDa protein were also adherent to HEp-2 cells, and anti-43-kDa protein antibody blocked adherence of 43-kDa protein-coated latex beads. These data suggest that the 43-kDa OMP functions as an adhesin in A. caviae.

Adhesins, Bacterial↗

Haemolysin occurrence among Aeromonas hydrophila, Aeromonas caviae and Aeromonas sobria strains isolated from different aquatic ecosystems.

A total of 909 Aeromonas spp. isolates from different aquatic ecosystems were tested for haemolysin production by both sheep and horse blood agar-plate assays and by rabbit erythrocytes in broth assay. A comparison of these different methods was undertaken in order to appreciate their capacity to evaluate the haemolytic activity of Aeromonas spp. isolated from aquatic ecosystems. The haemolytic activity was associated particularly with A. hydrophila and A. sobria (about 95% of strains), whereas A. caviae did not produce haemolysin (about 95% of strains). A method suitable for use in routine diagnostic microbiology laboratories is proposed for quantifying both groups of A. hydrophila/A. sobria and A. caviae in environmental water.

Aeromonas↗

Enumeration and confirmation of Aeromonas hydrophila, Aeromonas caviae, and Aeromonas sobria isolated from raw milk and other milk products in Northern Greece.

A total of 138 raw cow's and 57 raw ewe's milk samples; 80 pasteurized cow's milk samples; 39 Anthotyros cheese, 36 Manouri cheese, and 23 Feta cheese samples; and 15 rice pudding samples were examined for the presence and any countable population of Aeromonas species. Twenty-two (15.9%) of the 138 cow's milk samples analyzed were contaminated with A. hydrophila. In 13 of these samples, populations of 3.0x10(2) to 5.0x10(3) CFU/ml were counted in starch ampicillin agar (SAA). Eighteen cow's milk samples (13.0%) were contaminated with A. caviae, and in eight of these samples, populations of 2.0x10(2) to 3.0x10(3) CFU/ml were counted in SAA. Five cow's milk samples (3.6%) were contaminated with A. sobria, and in two of these samples, populations of 2.5x10(3) and 5.0x10(3) CFU/ml were counted in SAA. Eleven cow's milk samples (7.9%) were contaminated with other Aeromonas spp. not classified. Eight (14.0%) of the 57 ewe's milk samples analyzed were contaminated with A. hydrophila. In these samples, populations of 5.0x10(2) to 5.0x10(3) CFU/ml were counted in SAA. Six ewe's milk samples (10.5%) were contaminated with A. caviae, and populations of 1.5x10(2) to 1.0x10(3) CFU/ ml were counted in SAA. Two ewe's milk samples (3.5%) were contaminated with A. sobria, and populations counted in SAA were 5.0x10(2) and 1.0x10(3) CFU/ml. Four samples (7.0%) were contaminated with other Aeromonas spp. not classified. A. hydrophila was recovered in 4 (10.2%) and 3 (8.3%) of the Anthotyros and Manouri cheese samples analyzed, respectively, but no countable populations were noted in SAA. None of the pasteurized milk, Feta cheese, and rice pudding samples yielded Aeromonas spp. The results of this work indicate that motile Aeromonas are common in raw milk in Greece. Also, the presence of A. hydrophila in the whey cheeses Anthotyros and Manouri indicates that postprocessing contaminations of these products with motile Aeromonas may occur during production.

Aeromonas↗

[Virulence factors of clinical Aeromonas caviae isolates].

Aeromonas caviae, an ubiquitous aquatic organism, has long been considered to be of low pathogenicity, and its virulence mechanisms are still not clearly understood. Twenty-eight A. caviae isolates of clinical origin, most often monomicrobic, were identified in our university hospital over a four year period. Patients, mostly immunocompromised, were: eight diarrhoeal infants, 13 diarrhoeal adults, seven bacteraemic adults. Adults were frequently suffering from underlying intestinal malignancy, hepatobiliary disease, gastrectomy. Virulence factors were investigated. Adherence, studied by use of tissue culture HEp-2 cells, and staining of characteristic lateral flagella, were observed in diarrhoeal strains. Extracellular hemolytic activity was tested on rabbit erythrocytes suspensions at 25 and 37 degrees C. One blood culture isolate showed an important hemolytic activity at 25 degrees C, but none at 37 degrees C. Treatment with furin activated the aerolysin precursor and resulted in significant hemolysis at 37 degrees C, and fluid accumulation in rabbit ileal loops similar to that of A. hydrophila as control. The presence of the hemolysin gene was confirmed in this strain by PCR. In conclusion, A. caviae was shown to be a pathogen isolated from diarrhoea and bacteraemia in immunocompromised patients with malignancies and low gastric acidity as favouring factors. Virulence including the ability to adhere to cells and the production of lateral flagella was observed in diarrhoeal strains. The expression and the production of extracellular hemolytic activity and enterotoxicity at 37 degrees C depended on the activation of the pore forming toxin aerolysin precursor by furin. In vivo the protoxin is probably processed to its mature form by host proteases.

Adult↗

Purification and some properties of high-molecular-weight xylanases, the xylanases 4 and 5 of Aeromonas caviae W-61.

Aeromonas caviae W-61 produces multiple extracellular xylanases, the xylanases 1, 2, 3, 4, and 5 [Nguyen, V. D. et al., Biosci. Biotechnol. Biochem., 56, 1708-1712 (1993)]. Here we purified and characterized high-molecular-weight xylanases, the xylanases 4 and 5 from the culture fluids of the bacterium. The purified xylanases 4 and 5, which had molecular masses of 120 and 140 kDa, respectively, were endo-beta-1,4-xylanases with similar enzymatic properties except for trans-xylosidase activity. The xylanase 4 showed a prominent transxylosidase activity when xylotriose and xylotetraose were used as the substrates, while the xylanase 5 had little transxylosidase activity under the same conditions. Protein sequencing indicated that the xylanase 4 was a C-terminally-truncated xylanase 5, suggesting that the C-terminal truncation of the xylanase 5 may endow the enzyme with transxylosidase activity.

Aeromonas↗

Molecular properties and activity of a carboxyl-terminal truncated form of xylanase 3 from Aeromonas caviae W-61.

Aeromonas caviae W-61 produces five species of xylanases, xylanases 1, 2, 3, 4, and 5 [Nguyen, V.D. et al., Biosci. Biotechnol. Biochem., 56, 1708-1712 (1993) and Appl. Environ. Microbiol., 57, 445-449 (1991)]. While preserving a purified xylanase 3 preparation from A. caviae in solution at 4 degrees C, the xylanase 3 was found to be proteolyzed to give a truncated form with a smaller molecular mass than that of the intact one. It appears likely that the truncated form of xylanase 3 was produced in this particular purification experiment by the action of a contaminating protease. We isolated the truncated form of xylanase 3 (Xyn3tr), of which the C-terminal 102-residue segment is missing. By the chemical analysis of the N- and C-terminal amino acid residues of Xyn3tr and the DNA sequencing analysis of the xylanase 3 gene (xyn3), the N-terminal 398th proline residue of xylanase 3 was found to be the C-terminus of Xyn3tr. Xyn3tr had the activity to form xylotriose (X3), xylotetraose (X4), xylopentaose (X5), and xylohexaose (X6) as main final products from oat spelt xylan. In contrast, intact xylanase 3 released X6 and higher xylo-oligosaccharides as main products. Xylanase 3 hydrolysed X4 through X6. However, Xyn3tr had no activity towards X4 and X5. The recombinant Xyn3tr and recombinant xylanase 3 (XYN3) were purified homogeneously from the periplasmic space of E. coli harboring the plasmids pXYN3 and pXYN3tr, which include xyn3 and xyn3tr genes, respectively, and their enzymatic activities were measured. The cleavage patterns of oat spelt and xylo-oligosaccharides by XYN3tr were identical with that by intact Xyn3tr. Thus, we conclude that the C-terminal region comprising a 102-residue segment in xylanase 3 is involved in governing the molecular size of xylo-oligosaccharides cleaved from beta-1,4-xylan by the enzyme and in the hydrolytic activity towards X4 and X5.

Aeromonas↗

Purification and Properties of beta-1, 4-Xylanase from Aeromonas caviae W-61.

Aeromonas caviae W-61, which was isolated from water samples at the Faculty of Agriculture, Tohoku University, produced beta-1, 4-xylanase (1,4-beta-d-xylan xylanohydrolase; EC 3.2.1.8) extracellularly. The xylanase was purified to homogeneity by using DEAE-Sephadex A-50, CM-Sephadex C-50, and Sephadex G-100 column chromatographies. The molecular weight of the purified enzyme was estimated to be 22,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of the enzyme was 9.2. The optimal pH and temperature for the activity of the enzyme were 7.0 and 55 degrees C, respectively. The enzyme was stable at pH 7.0 at temperatures of up to 50 degrees C. As enzymatic products, various xylo-oligosaccharides such as xylobiose, xylotriose, xylotetraose, and xylopentaose were formed, and only a small amount of xylose was detected. The purified enzyme did not hydrolyze starch, cellulose, carboxymethylcellulose, or beta-1, 3-xylan.

Journal Article↗

Aeromonas eucrenophila species nova Aeromonas caviae a later and illegitimate synonym of Aeromonas punctata.

Aeromonas caviae is a later and illegitimate synonym of Aeromonas punctata. In order to secure continuity in literature, it is proposed to use the trivial term "caviae type" in connection with the species name A. punctata temporarily especially for strains isolated in connection to diarrhoea in children. Aeromonas eucrenophila sp. nov. is described, type strain in NCMB 74. The new aerogenic psychrotrophic Aeromonas species is separated genotypically from Aeromonas hydrophila and Aeromonas punctata (caviae type).

Aeromonas↗

Chronic diarrhea due to a single strain of Aeromonas caviae.

Over a period of 17 months, Aeromonas caviae was cultured 15 times as the sole enteropathogen from the feces of a man who had developed chronic diarrhea after traveling to Turkey. Determination of rRNA gene restriction patterns confirmed that the seven isolates of Aeromonas caviae studied were identical (hybridization group [HG]4). After therapy with ciprofloxacin for four weeks, the patient was culture negative for the original isolate, and six months later a novel strain of Aeromonas media with a different ribopattern (HG 5A) was isolated from the feces of the patient. The patient responded again, both clinically and bacteriologically, to a four-week course of ciprofloxacin and has remained asymptomatic since then.

Aeromonas↗

Aeromonas caviae: an enteric pathogen?

Of the Aeromonas spp. isolated routinely in our laboratory from human feces, about two thirds represent Aeromonas caviae. In contrast to Aeromonas hydrophila and Aeromonas sobria, this species has been considered to be of little enteropathogenic significance due to the absence of known virulence factors. The clinical data presented here suggest a clinical significance of A. caviae, at least in some cases. As yet nothing is known about the pathogenic mechanisms involved.

Adolescent↗

Cloning and primary structure of the chiA gene from Aeromonas caviae.

The chiA gene from Aeromonas caviae encodes an extracellular chitinase, 865 amino acids long, that shows a high degree of similarity to chitinase A of Serratia marcescens. Expression in Escherichia coli yielded an enzymatically active protein from which a leader sequence was removed, presumably during transport of the enzyme across the cell membrane.

Aeromonas↗