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Immunochemical analysis and possible biological role of an Aeromonas hydrophila surface array protein in septicaemia.

The biochemical, immunological, and biological properties of an S layer purified from an Aeromonas hydrophila strain (AH-342) involved in a case of bacteraemia were investigated. The S layer selectively removed from the cell surface was composed of a single acidic (pI 4.56) protein subunit (surface array protein, SAP) with a molecular mass of approximately 52 kDa. Amino acid analysis of this 52 kDa protein indicated a molecule composed of 498 amino acids with 46% hydrophobic residues. No cysteine residues were detected. The first 35 residues of the N-terminus were sequenced by Edman degradation; only 4-24% homology was noted between this sequence and those previously published for SAPs of Aeromonas salmonicida (A450) and a strain of A. hydrophila (TF7) originally isolated from a moribund fish. Polyclonal antibodies raised against AH-342 SAP were genospecific, reacting only against S layers produced by A. hydrophila strains and not those from Aeromonas veronii. Acute serum from the bacteraemic patient from whom AH-342 was isolated reacted strongly with the SAP of AH-342 in immunoblot studies. Purified SAP, when intraperitoneally co-inoculated with SAP- strains of A. hydrophila into Swiss-Webster mice, could reduce the 50% lethal dose by approximately 30-70 fold. The results suggest that the SAP of A. hydrophila strains may play an important role in systemic dissemination after invasion through the gastrointestinal mucosa.

Aeromonas hydrophila↗

Aeromonas cytotonic enterotoxin cross reactive with cholera toxin.

Isolation by affinity chromatography from crude culture filtrate of Aeromonas sobria of protein that cross reacted with cholera toxin (CT) revealed a toxin that produced fluid accumulation in rat ileal loops and in infant mice and caused rounding of Y1 adrenal cells. All these activities were neutralised by antiserum to CT. There was no haemolytic or cytotoxic activity associated with this CT-cross reactive cytotonic enterotoxin. CT-cross reactive material detected in enzyme linked immunosorbent assay (ELISA) was produced by 25% of Aeromonas isolates from faeces of children with or without diarrhoea-26% of A. sobria, 20.0% of A. hydrophila and 24% of A. caviae tested gave positive ELISA results. Most strains that produced this cytotonic enterotoxin but no cytotoxic enterotoxin were isolated from children without diarrhoea. Toxin preparations from Aeromonas spp. that completely inhibited adenosine-5'-diphosphate-induced platelet aggregation, an effect related to elevation of intracellular cAMP, were, with one exception, cross reactive with CT in ELISA.

Aeromonas↗

Occurrence of Aeromonas species and Plesiomonas shigelloides in patients with and without diarrhoea in Lagos, Nigeria.

The prevalence of Aeromonas spp. and Plesiomonas shigelloides was determined in patients attending the enteric laboratory of the Department of Medical Microbiology and Parasitology, Lagos University Teaching Hospital, Nigeria. During the 12-month study (October 1986-September 1987), Aeromonas spp. were isolated from 53 (2.26%) of 2350 patients with diarrhoea and only 2 (0.4%) of 500 patients without diarrhoea (p less than 0.01). Similarly, P. shigelloides was isolated from 16 (0.68%) patients with diarrhoea and none of the controls (p greater than 0.05). The seasonality, age and sex distribution of diarrhoea associated with Aeromonas spp. and P. shigelloides in this study were similar to those of diarrhoea associated with other recognised enteropathogens in Nigeria. Both species may play a role in the aetiology of acute diarrhoeal disease in that environment.

Adolescent↗

Enterotoxigenicity, hemolytic activity and antibiotic resistance of Aeromonas spp. isolated from freshwater prawn marketed in Dhaka, Bangladesh.

Aeromonas spp. were isolated from gills, swimmerets, eggs, stomachs and ventral muscles of freshwater prawns (Macrobrachium malcolmsonii) available in the local fish market of Dhaka, Bangladesh. The density of Aeromonas spp. on these different body parts of the prawn samples ranged from 1.1 +/- 0.2 x 10(4) to 1.5 +/- 0.16 x 10(7) cfu per gram. The viable counts of aeromonads, fecal coliforms (FC) and Escherichia coli gradually increased in prawn samples when stored at 4 C. At -20 C, the viable counts gradually decreased and became zero on the 12th day of storage. The isolation of A. sobria (56%) was more frequent than that of A. hydrophila (31%) and A. caviae (13%). In the rabbit ileal loop (RIL) model, fluid accumulation induced by live cultures and cell-free culture filtrates of 11 strains ranged from 0.5 to 1.5 and 0.5 to 1.7 ml/cm of gut, respectively. Of 11 enterotoxigenic strains, 7 were A. sobria and 4 were A. hydrophila. Enterotoxigenicity correlated with hemolytic activity on blood agar. All enterotoxigenic strains were uniformly sensitive to chloramphenicol and gentamicin and resistant to novobiocin and vancomycin. Isolation of enterotoxigenic and antibiotic-resistant Aeromonas from these prawn samples indicates possible public health problems for their handlers as well for raw prawn consumers.

Aeromonas↗

Production of exotoxins by Aeromonas spp. at 5 degrees C.

The ability of 60 strains of Aeromonas to produce enterotoxin and haemolysin after cultivation at 5 degrees C for 7-10 d was investigated. The strains were isolated from lamb meat, offal, carcasses and faeces, and had previously been tested for their ability to produce these exotoxins at 37 degrees C. The results showed that some strains of Aeromonas hydrophila and A. sobria were capable of producing enterotoxin and haemolysin at 5 degrees C, but none of the A. caviae strains tested produced these two factors. Of the 30 A. hydrophila strains investigated 25 and 27 were enterotoxigenic and haemolytic respectively. Likewise, of the 24 A. sobria strains investigated 16 and 18 were enterotoxigenic and haemolytic respectively. The results indicate that certain strains of Aeromonas species, in particular A. hydrophila and A. sobria, are of potential public health significance in meats stored at refrigeration temperature.

Aeromonas↗

Typing of Aeromonas strains from patients with diarrhoea and from drinking water.

Aeromonas strains (187) from human diarrhoeal stools and from drinking water (263) in The Netherlands were typed by three different methods. Biotyping alone was found to be of little value for epidemiological studies because 84% of all strains belonged to only 10 biotypes. Common biotypes could be further differentiated by serotyping. Gas-liquid chromatography of cell wall fatty acid methyl esters (FAME) was useful for species identification as well as for typing: 86% of all strains could be identified to the species level, and within this group 92% of all identifications corresponded with the biotype. Cluster analysis and principal component analysis of FAME profiles could be used for comparison of strains from different sources and gave the same general conclusions as bio- and serotyping. There was little overall similarity between Aeromonas strains from human (diarrhoeal) faeces and from drinking water, differences being most pronounced for Aeromonas caviae and least for A. sobria.

Aeromonas↗

Detection of Aeromonas hydrophila in food with an enzyme-linked immunosorbent assay.

A microtitration plate, antibody capture, enzyme-linked immunosorbent assay was developed for the detection of Aeromonas hydrophila serotype O: 11 (highly virulent strains). The assay utilizes a detector antibody which shows no cross-reactions with Aeromonas strains other than serotype O: 11 or non-Aeromonas competing organisms. The detector antibody is mixed with the sample and incubated for 1 h, microcentrifuged and the supernatant fluid (unadsorbed antibody) titred in a microtitre plate coated with A. hydrophila cells from serotype O: 11. All the A. hydrophila strains from serotype O: 11 tested reacted strongly with the detector antibody. Also by culturing and performing the immunoassay with the detector antibody we established and quantified the presence of A. hydrophila O: 11 in different foods.

Aeromonas hydrophila↗

Differences in production of several extracellular virulence factors in clinical and food Aeromonas spp. strains.

Production of several extracellular virulence factors (lipase, protease and haemolysin) was compared in 15 Aeromonas spp. isolated from faeces of patients with Aeromonas-associated gastroenteritis and 81 strains isolated from food. Strains from food did not show differences in production of these factors when compared with strains isolated from faeces. However, if strains were considered in relation to autoagglutination (AA) character, the AA+ differed from AA- strains in lipase and protease production. Supernatant fluids of AA+ food and human strains showed 2.5-fold more protease production than that observed in AA- strains. These two characteristics of certain Aeromonas strains could be related with the more virulent capacity.

Aeromonas↗

Differentiation of Aeromonas genomospecies using random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR).

Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to investigate the differentiation of the genus Aeromonas at the genomospecies level. Of 20 primers evaluated, six produced profiles which contained multiple bands capable of differentiating the genomospecies. These six primers were also used in RAPD-PCR analyses of clinical and environmental isolates of the different genomospecies. In most cases, each strain gave a unique fingerprint, illustrating genetic heterogeneity at the genomospecies level. However, some homogeneity in fragment sizes was seen among strains within a genomospecies which was not apparent in strains from different genomospecies. This study therefore complements and supports the current classification of Aeromonas into genomospecies. These results also show that RAPD-PCR has the potential to differentiate between the genomospecies of Aeromonas.

Aeromonas↗

Septic arthritis due to Aeromonas hydrophila: case report and review of the literature.

Aeromonas hydrophila is a rare human pathogen, and worldwide, soft tissue infections following water-related injuries are the most common. However, septic arthritis due to A. hydrophila remains uncommon with only seven cases previously reported in the English literature. In this report, we describe the important clinical features, microbiological findings and management of severe septic arthritis of the knee due to A. hydrophila in a healthy 13-year-old girl following an injury sustained in a private fresh water lake. A review of seven previously reported cases of septic arthritis due to A. hydrophila and the present case suggests that the infection commonly affected the knee and the meta-/intercarpal-phalangeal joint and was frequently rapidly progressive following trauma in fresh water and or associated with leukaemia. Second and third generation cephalosporins, gentamicin, trimethoprim, ciprofloxacin and appropriate orthopaedic management should be promptly instituted. After chemical treatment and chlorination, fresh water becomes free of coliforms, but A. hydrophila persists more compared with the other strains of aeromonas, namely A. sobria and A. caviae. Seawater injuries, unlike freshwater injuries, are not usually associated with aeromonas infections. Further workup on the mechanisms of A. hydrophila resistance to chlorination could probably yield useful information in achieving new procedures of preventing and controlling such infections in public and private fresh water recreational facilities.

Adolescent↗

Carp erythrodermatitis (CE) due to an Aeromonas hydrophila infection. Casuistic and experimental results.

In November 1987 high losses of carp (Cyprinus carpio) with the main symptom of skin ulcera were observed in a farm in northern Greece. Sixty-six isolates of bacteria, characterized mainly as Aeromonas hydrophila or Pseudomonas spp. could be isolated from lesions of diseased fish. Transmission experiments with these isolates using mirror carp showed that Aeromonas hydrophila strains induced identical clinical and pathological pictures after intra- or subcutaneous injection. Extracts of these Aeromonas hydrophila isolates, as well as a supernatant of culture bouillon were toxic for carp and mice, indicating the presence of endo- and exotoxins. The results prove that carp erythrodermatitis (CE) may be caused by different bacteria, mainly including A. hydrophila.

Aeromonas↗

Computer aided assignment of motile Aeromonas strains to genospecies level by standard biochemical tests.

A procedure is given for differentiation of motile Aeromonas spp. Based on nine to 12 biochemical properties it allows an assignment of isolated Aeromonas strains first to phenotypes and, in the second step, to hybridization groups (HGs) 1-13 as well as to the genotypically defined species Aer. allosaccharophila and Aer. encheleia. The computer aided classification is carried out according to the principles of numerical taxonomy. Using this differentiation scheme 23 Aeromonas strains, which were isolated from food, and two reference strains were speciated. A total of 16 strains were assigned to Aer. hydrophila, six strains to Aer. caviae and one strain to Aer. sobria. Of the 16 Aer. hydrophile strains, 11 were attached to HG 1, one to HG 2 and four could not be classified clearly as HG 3 or HG 1. All six Aer. caviae isolated were assigned to HG 5 and the Aer. sobria strain was ranked among HG 8. The reference strains were assigned to the same genotypes by this method than by DNA-DNA hybridization assays.

Aeromonas↗

Cellulitis owing to Aeromonas hydrophilia: treatment with hyperbaric oxygen.

Aeromonas hydrophilia, a Gram-negative facultative anaerobe, is a rare cause of cellulitis. We review the literature and report a case of Aeromonas hydrophilia cellulitis which failed to respond to the conventional management of surgical debridement and antibiotics. Hyperbaric oxygen therapy was used successfully to control the infection. As far as we are aware this is the first case in world literature of Aeromonas hydrophilia cellulitis treated with hyperbaric oxygen.

Aeromonas hydrophila↗

Pili and the interaction of Aeromonas species with human peripheral blood polymorphonuclear cells.

The interaction of differentially piliated Aeromonas strains expressing pili of two broadly different morphologic types (short, rigid (S/R) and/or long, wavy (L/W)) with human peripheral blood mononuclear leukocytes (PMN) was investigated to determine whether host defense cells might exert a selective pressure on pili expression in vivo accounting for the different pili phenotypes of clinical and environmental strains. A majority of Aeromonas veronii biotype sobria strains from water (6/6) and faeces (8/11) readily associated with PMN (> 60% PMN with adherent and/or internalised bacteria), irrespective of their degree, or predominant type, of piliation. Rigid pili of Aeromonas species did not promote interaction with PMN. However, the majority (55%) of strains which interacted well with PMN were adherent to HEp-2 cells. Interaction with PMN is unlikely to be the reason few S/R pili are seen on faecal strains, but it may be a selective pressure on L/W adhesive pili, or other OMP adhesins, resulting in the shedding of strains which have lost critical adhesins.

Aeromonas↗

Potential virulence and antimicrobial susceptibility of Aeromonas popoffii recovered from freshwater and seawater.

Aeromonas popoffii is the most recent species within the genus Aeromonas described from freshwater. In our study this species was also recovered from this habitat and for the first time from seawater. Most of the virulence factors known in Aeromonas spp. (aerolysin/hemolysin, serine protease, lipases and DNases) were highly prevalent in this species. Third-generation cephalosporins and quinolones were the most active antimicrobial agents against A. popoffii.

Aeromonas↗

Signature region within the 16S rDNA sequences of Aeromonas popoffii.

To identify a group of eight Aeromonas strains of our collection showing ribotyping patterns similar to those described for the species Aeromonas popoffii, 16S rRNA gene sequence analysis was performed. Results were in agreement with the DNA binding values, and allowed the identification of a 'signature region' differentiating the A. popoffii strains from all other members of the genus Aeromonas.

Aeromonas↗

Distribution of cphA or related carbapenemase-encoding genes and production of carbapenemase activity in members of the genus Aeromonas.

The prevalence of the cphA gene or related carbapenemase-encoding genes was investigated in 114 Aeromonas strains belonging to the six species of major clinical interest. A species-related distribution of cphA-related sequences was observed. Similar sequences were found in A. hydrophila, A. veronii bv. sobria, A. veronii bv. veronii, and A. jandaei, but not in A. caviae, A. trota, or A. schubertii. However, a single A. caviae strain (of 62 tested) was found carrying cphA-related sequences, suggesting the possibility of the horizontal transfer of this gene to species which normally do not carry it. Production of carbapenemase activity was detectable in 83% of the hybridization-positive strains but in none of the hybridization-negative ones. When it was present, carbapenemase activity was always inhibitable by EDTA. Either carbapenemase-producing or not, Aeromonas strains appeared to be susceptible to imipenem when in vitro susceptibility testing was performed with inocula of conventional size (10(5) CFU), for which MICs were always < or = 1 microgram/ml. With a larger inoculum (10(8) CFU), the MICs for carbapenemase-negative strains always remained < or = 1 microgram/ml, while those for carbapenemase-producing strains were always > or = 4 micrograms/ml, being usually higher than the breakpoint for susceptibility. The present results indicate that the production of metallocarbapenemase activity, apparently encoded by cphA homologs, is widespread among some of the Aeromonas species of clinical interest (A. hydrophila, A. veronii bv. sobria, A. veronii bv. veronii, and A. jandaei) and that imipenem MICs for carbapenemase-producing strains are subjected to a relevant inoculum size effect.

Aeromonas↗

Antimicrobial susceptibilities of Aeromonas spp. isolated from environmental sources.

Aeromonas spp. are ubiquitous aquatic bacteria that cause serious infections in both poikilothermic and endothermic animals, including humans. Clinical isolates have shown an increasing incidence of antibiotic and antimicrobial drug resistance since the widespread use of antibiotics began. A total of 282 Aeromonas pure cultures were isolated from both urban and rural playa lakes in the vicinity of Lubbock, Texas, and several rivers in West Texas and New Mexico. Of these, at least 104 were subsequently confirmed to be independent isolates. The 104 isolates were identified by Biolog and belonged to 11 different species. The MICs of six metals, one metalloid, five antibiotics, and two antimicrobial drugs were determined. All aeromonads were sensitive to chromate, cobalt, copper, nickel, zinc, cefuroxime, kanamycin, nalidixic acid, ofloxacin, tetracycline, and sulfamethoxazole. Low incidences of trimethoprim resistance, mercury resistance, and arsenite resistance were found. Dual resistances were found in 5 of the 104 Aeromonas isolates. Greater numbers of resistant isolates were obtained from samples taken in March versus July 2002 and from sediment versus water. Plasmids were isolated from selected strains of the arsenite- and mercury-resistant organisms and were transformed into Escherichia coli XL1-Blue MRF'. Acquisition of the resistance phenotypes by the new host showed that these resistance genes were carried on the plasmids. Mercury resistance was found to be encoded on a conjugative plasmid. Despite the low incidence of resistant isolates, the six playa lakes and three rivers that were sampled in this study can be considered a reservoir for antimicrobial resistance genes.

Aeromonas↗