Occurrence of aeromonas hydrophila and aeromonas shigelloides in feces.
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Ribotyping was used to study the epidemiology of Aeromonas associated gastro-enteritis in young children. Ribotyping patterns of 29 Aeromonas strains (16 Aeromonas caviae, 8 Aeromonas hydrophila, 3 Aeromonas eucrenophila, 1 Aeromonas veronii, and 1 Aeromonas encheleia) isolated from primary stool cultures of sick children were compared using the GelCompare software with patterns of 104 strains (39 Aeromonas eucrenophila, 29 Aeromonas caviae, 11 Aeromonas encheleia, 10 Aeromonas hydrophila, 6 Aeromonas bestiarum, 3 Aeromonas veronii, 3 Aeromonas popoffii and 3 Aeromonas media) isolated from their household environment in order to investigate the route of transmission of these bacteria. Fifteen strains (approximately 47%) isolated from stool cultures of patients showed the same riboprofile as strains found in contacts or environment. In particular, three strains isolated from patients shared the same riboprofile with strains found in their domestic environment. The wide diffusion of potentially pathogenic Aeromonas strains in our household samples, and the high rate of asymptomatic carriers among family members, suggested that predisposing factors of the host could make children prone to an Aeromonas-related intestinal disease.
OBJECTIVES: The objectives of this study were to elucidate the clinical manifestations of Aeromonas infections and the association of putative virulence genes with clinical invasiveness. METHODS: 116 consecutive clinical Aeromonas isolates collected from July 1999 to June 2001 in a medical center in southern Taiwan were included. All isolates were identified by biochemical phenotyping and their genomic sequences encoding eight putative virulence factors, including cytolytic enterotoxin (AHCYTOEN), aerolysin (aerA), hemolysin (hlyA), heat-labile enterotoxin (alt), heat-stable enterotoxin (ast), and components of type III secretion system (ascV, aexT and ascF-ascG) were analyzed using polymerase chain reaction and colony blot hybridization. The association of clinical diseases of the patients with the putative virulence genes in the isolates was analyzed. RESULTS: Sixty-two percent of Aeromonas isolates caused clinically evident infections, of which the major clinical manifestations were primary bacteremia (40%), followed by soft tissue infections (27%), and hepatobiliary tract infections (15%). Liver cirrhosis (36%), malignancy (25%), and hepatobiliary diseases (13%) were the major underlying diseases in patients with Aeromonas bacteremia. The majority (64%) of patients with Aeromonas hepatobiliary infections had underlying hepatobiliary diseases, whereas 71% of those with soft tissue infections had antecedent water- or soil-related injuries. The crude fatality rate for Aeromonas infections was 26%. Aeromonas hydrophila complex was the most common (52%) of the three major complex groups investigated, followed by Aeromonas sobria complex (24%) and Aeromonas caviae complex (23%). None of the eight putative virulence factors was associated with invasiveness or bacteremia. CONCLUSIONS: Primary bacteremia, soft tissue infections, and hepatobiliary tract infections are the three major clinical manifestations of invasive Aeromonas infections in southern Taiwan. This study found no association between the presence of AHCYTOEN, aerA, hlyA, alt, ast, ascV, aexT or ascF-ascG genes in Aeromonas isolates and the development of extra-intestinal infections or bacteremia.
AIMS: This study used restriction fragment length polymorphism (RFLP) with Aeromonas-specific primers to identify species of Aeromonas and to investigate their distribution in a trout farm and stream. METHODS AND RESULTS: In January, May, August and November 2000, presumptive Aeromonas species were recovered from a farm and a sedimentation pond in a fish farm and stream, and identified by PCR-RFLP analysis with Aeromonas-specific primers. The specificity of Aeromonas-specific primers and the suitability of PCR-RFLP analysis for identifying Aeromonas spp. were confirmed with fatty acid methyl esters (FAMEs) and 16S rDNA sequencing analyses, respectively. Levels of Aeromonas spp. sampled in May and August were higher than in January and November at all sampling sites. Aeromonas salmonicida was the dominant species in January and November, and the proportion of pathogenic species (Aer. hydrophila, Aer. caviae and Aer. veronii) increased in May and August. CONCLUSIONS: PCR-RFLP analysis with Aeromonas-specific primers is a rapid and reliable method for identifying widely distributed Aeromonas spp. from environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: To minimize human health risk, monitoring the levels and species composition of Aeromonas in fish farm is advisable.
We investigated the ability of a recently developed genomic fingerprinting technique, named AFLP, to differentiate the 14 currently defined DNA hybridization groups (HGs) in the genus Aeromonas. We also determined the taxonomic positions of the phenospecies Aeromonas allosaccharophila, Aeromonas encheleia, Aeromonas enteropelogenes, and Aeromonas ichthiosmia, which have not been assigned to HGs yet. A total of 98 Aeromonas type and reference strains were included in this study. For the AFLP analysis, the total genomic DNA of each strain was digested with restriction endonucleases ApaI and TaqI. Subsequently, restriction fragments were selectively amplified under high-stringency PCR conditions. The amplification products were electrophoretically separated on a polyacrylamide gel and visualized by autoradiography. Following high-resolution densitometric scanning of the resulting band patterns, AFLP data were further processed for a computer-assisted comparison. A numerical analysis of the digitized fingerprints revealed 13 AFLP clusters which, in general, clearly supported the current Aeromonas taxonomy derived from DNA homology data. In addition, our results indicated that there is significant genotypic heterogeneity in Aeromonas eucrenophila (HG6), which may lead to a further subdivision of this species. A. allosaccharophila and A. encheleia did not represent a separate AFLP cluster but were found to be genotypically related to HG8/10 and HG6, respectively. In addition, the results of the AFLP analysis also confirmed the phylogenetic findings that A. enteropelogenes and A. ichthiosmia are in fact identical to Aeromonas trota (HG13) and Aeromonas veronii (HG8/10), respectively. The results of this study clearly show that the AFLP technique is a valuable new high-resolution genotypic tool for classification of Aeromonas species and also emphasize that this powerful DNA fingerprinting method is important for bacterial taxonomy in general.
A hospital based study was carried out in the Department of Microbiology, Tribhuvan University Teaching Hospital, Institute of Medicine with the aim to initiate the isolation and identification of Aeromonas spp. from the stool samples of gastroenteritic patients and to determine the prevalence of Aeromonas spp. in other clinical samples and water. Altogether 293 samples were investigated that include 172 stool samples, 60 pus/wound swabs, 20 body fluids and 41 water samples. The samples were collected and processed by standard microbiological techniques in order to isolate Aeromonas. Ampicillin blood agar (20 microg/ml) was used as selective medium for the isolation of Aeromonas. The specimen prevalence rate of Aeromonas spp. in stool was found to be 5.2% and the A. hydrophila (55.5%) was the predominant species followed by A. caviae (33.3%) and A. sobria (11.1%) in the stool samples. Likewise, 3.3% of pus sample showed positive growth of A. hydrophila. Aeromonas was not detected in any of the body fluids. Aeromonas spp. were isolated from 58.5% of water samples obtained from different hospitals. The commonest species was A. hydrophila (62.5%) followed by A. caviae (20.8%) and A. sobria (16.7%). In vitro susceptibility testing showed that the aminoglycosides and fluroquinolones were the effective antibiotics against Aeromonas. It was found that 88.9% of Aeromonas spp. isolated from stool samples were sensitive to gentamicin, 77.8% to ciprofloxacin, norfloxacin, tetracycline and ceftriaxone and 66.6% to nalidixic acid whereas cent percent Aeromonas spp. from water samples were sensitive to gentamicin, ciprofloxacin, norfloxacin, tetracycline and ceftriaxone and 75.0% to nalidixic acid. The enteric Aeromonas isolates were more resistant to antimicrobial agents than the water isolates.
A total of 82 strains of presumptive Aeromonas spp. were identified biochemically and genetically (16S rDNA-RFLP). The strains were isolated from 250 samples of frozen fish (Tilapia, Oreochromis niloticus niloticus) purchased in local markets in Mexico City. In the present study, we detected the presence of several genes encoding for putative virulence factors and phenotypic activities that may play an important role in bacterial infection. In addition, we studied the antimicrobial patterns of those strains. Molecular identification demonstrated that the prevalent species in frozen fish were Aeromonas salmonicida (67.5%) and Aeromonas bestiarum (20.9%), accounting for 88.3% of the isolates, while the other strains belonged to the species Aeromonas veronii (5.2%), Aeromonas encheleia (3.9%) and Aeromonas hydrophila (2.6%). Detection by polymerase chain reaction (PCR) of genes encoding putative virulence factors common in Aeromonas, such as aerolysin/hemolysin, lipases including the glycerophospholipid-cholesterol acyltransferase (GCAT), serine protease and DNases, revealed that they were all common in these strains. Our results showed that first generation quinolones and second and third generation cephalosporins were the drugs with the best antimicrobial effect against Aeromonas spp. In Mexico, there have been few studies on Aeromonas and its putative virulence factors. The present work therefore highlights an important incidence of Aeromonas spp., with virulence potential and antimicrobial resistance, isolated from frozen fish intended for human consumption in Mexico City.
The performance of four media for the isolation of Aeromonas strains from stool specimens, the importance of ampicillin-susceptible Aeromonas strains in the selection of culture media, and the usefulness of beta-hemolysis in screening blood-containing media for Aeromonas strains were evaluated in two phases. In the first phase, 36 of 1,672 stool specimens yielded Aeromonas isolates. Ninety-seven percent of the isolates were detected on blood agar containing 20 micrograms of ampicillin per ml (ABA), and 47% were detected on MacConkey agar containing 100 micrograms of ampicillin per ml and 1% Tween 80. In the second phase of the study, 43 of 1,924 stool specimens yielded Aeromonas isolates. Fifty-one percent of the isolates were detected on blood agar and on modified cefsulodin-Irgasan-novobiocin agar, and 84% were detected on ABA. The combination of ABA and modified cefsulodin-Irgasan-novobiocin agar provided 100% recovery of the Aeromonas isolates encountered. All of the Aeromonas isolates detected on blood agar were also detected on ABA, and 89% of the Aeromonas isolates detected on these media were beta-hemolytic. These results suggest that ABA is superior to the other media evaluated for the isolation of Aeromonas strains from stool specimens, but optimal recovery of the organism may require the use of more than one medium. The results also suggest that the occurrence of ampicillin-susceptible strains is not a limitation on the use of ABA, but at least 10% of Aeromonas isolates will be missed if beta-hemolysis is used to screen ABA plates for these organisms.
OBJECTIVE: Aeromonas septicaemia complicating cirrhosis is not a common infectious disease. To enhance the knowledge in this aspect, we analysed the clinical features, pathogenetic factors, resistance, treatment and prognosis of Aeromonas septicaemia in 50 cases of hepatic cirrhosis. METHODS: We cultured the bacteria from infected patients with BacT/Alert120 automation instrument made in AKsu and identified the bacteria with the Vitek-AMS60 made in Biomerieux company. We then tested the susceptibility of Aeromonas to 13 antimicrobial agents. RESULTS: A total of 50 cases of Aeromonas septicaemia occurred in severe hepatic cirrhosis. The majority of them had severe complications. Aeromonas hydrophila was the most common species isolated (52.0%). Nosocomial infection was the predominant way of infection. The major clinical manifestations of Aeromonas septicaemia were fever (100%), chill (64.0%), abdominal pain (60.0%), diarrhoea (32.0%) and shock (24.0%). The susceptive rate of Aeromonas to third generation cephalosporin, quinolones and aminoglycoside antibiotics was more than 80%. The cure rate and mortality attributed to Aeromonas septicaemia after treating with third generation cephalosporin, levofloxacin and their combination were 64.3%, 75.0%, 57.1% and 28.6%, 12.5%, 35.7% respectively. CONCLUSION: Aeromonas septicaemia tends to befall patients with severe hepatic cirrhosis and causes a rapidly fatal outcome. Aeromonas should be considered an important pathogen for septicemia in patients with liver cirrhosis. It is suggested that we should emphasize the clinical features and laboratory diagnosis so as to have proper antimicrobial treatment.
BACKGROUND AND PURPOSE: Aeromonas spp. often cause infections in immunocompromised patients. To specifically understand the clinical features of Aeromonas bacteremic adults with hematologic malignancies, we investigated the demographic, clinical and microbiologic characteristics of Aeromonas bacteremia in this patient population. METHODS: Retrospective study performed in a tertiary medical center in southern Taiwan, in which adults with hematologic malignancies suffered from Aeromonas bacteremia admitted between 1995 and 2003 were included for study. RESULTS: There were 45 episodes of Aeromonas bacteremia in 41 adults with hematologic malignancies. Episodes of Aeromonas bacteremia which occurred at least 2 months apart were counted as separate cases in the analysis. A total of 30 men and 15 women (mean age: 53.2 years), with 4 patients experiencing 2 episodes, was included. The 3 leading underlying hematologic malignancies were acute myelogenous leukemia (37.8%), myelodysplastic syndrome (26.7%) and non-Hodgkin's lymphoma (17.8%). No cluster of Aeromonas bacteremia was found during the study period. Twenty nine (64.4%) of the 31 patients with nosocomial Aeromonas bacteremia had received recent antineoplastic chemotherapy. The 3 leading clinical manifestations were fever (88.9%), septic shock (40%), and altered consciousness (26.7%). Eleven (24.4%) episodes of bacteremia were polymicrobial. Sixteen (35.6%) patients died within 14 days of onset of bacteremia. The mean duration from sampling blood for culture to death was 3.81 days. Altered consciousness (odds ratio, 8.999; 95% confidence interval, 1.787-45.33; p=0.008) was the only independent prognostic factor for mortality. High resistance rates (11.1% to piperacillin and 35.6% to imipenem) among Aeromonas isolates were also noted. CONCLUSION: In febrile patients with hematologic malignancies and suspected Aeromonas infections, particular attention to the development of alteration of consciousness is needed as it is an independent risk factor for mortality.
Seasonal dynamics of motile Aeromonas in a treated urban effluent and in natural seawater along the Sfax coast (Mediterranean sea, Tunisia) were measured over a year concurrently with seven environmental factors, and compared with those of faecal coliforms. Counts for Aeromonas from a standard plate count method, ranged from 1.48 x 10(5)CFU.100 ml(-1) to 2.2 x 10(8)CFU.100 ml(-1) in the effluent and from 7.9 x 10(3)CFU.100 ml(-1) to undetectable level in the surface marine waters. Contrary to faecal coliforms, the Aeromonas dynamics exhibited a seasonal distribution in seawater which was inverse of the seasonal distribution in the sewage: From the end of November 1998 to April 1999 (cold period), Aeromonas counts increased in the treated effluent, while it decreased very rapidly in seawater. From May to October (warm period), Aeromonas abundance decreased in the effluent but showed an increasing fluctuating trend in the marine waters with a maximum in late summer/early autumn when the temperatures were around 22-23 degrees C. Multiple correlation and regression analyses suggest, by the coefficient of determination (R(2)), that 42% of variance in Aeromonas number changes in the treated effluent, may be explained by only turbidity, radiation and Aeromonas density in the previous sample, while 37% of variance in marine ecosystem were explained by radiance and conductivity. Furthermore, the t statistics and their p values and the coefficient of partial determination (r(2)) indicated that radiance contributed the most (r(2)=0.3184, t=-3.2, p=0.0041) to the dynamics of motile Aeromonas in seawater, when combined with conductivity. The models relevant for changes in faecal coliforms abundance incorporated turbidity, radiance in the effluent and conductivity, pH, radiance, turbidity in coastal marine environment. These models explain 66% and 73% of the observed cell number fluctuation, with turbidity (r(2)=0.529, t=5.08, p=0.0001) and conductivity (r(2)=0.5407, t=4.97, p=0.0001) as dominant factors in the multivariate model proposed, respectively, for the two sampling sites. The results presented here suggest that the combination of negative effects of sunlight and conductivity in natural seawater mainly affects the colony-forming capacity and make the motile Aeromonas nonrecoverable during cold months.
The significance of Aeromonas spp. as potential enteric pathogens was evaluated in a cohort of 187 infants aged 3-18 months during a 16-week summer period. Aeromonas spp. were isolated from 14 of the 196 (7.1%) diarrhoeal episodes detected and from eight (5.2%) of 153 samples from paired asymptomatic infants. Carriage of bacterial enteropathogens excluding Aeromonas spp. was detected in a high proportion (23%) of the asymptomatic children. Almost all of the seven isolates of Aeromonas sobria were enterotoxigenic, invasive and beta-haemolytic. In contrast, none of the seven Aeromonas caviae strains had these virulence-associated characteristics. The only isolate of Aeromonas hydrophila produced cytotoxic enterotoxin and was invasive. Plasmid analysis of selected strains did not correlate with these two properties or with antibiotic resistance. Nevertheless, the latter was found in an important proportion of the isolates. The diarrhoeal episodes, in which Aeromonas spp. were detected, lasted significantly longer, i.e. 17.2 days when the strains were invasive and/or toxigenic as compared with 4.3 days (P less than 0.001) in patients harbouring strains lacking both traits. These results reinforce the need to characterise virulence determinants before assigning any pathogenic role to Aeromonas spp. isolated from faecal specimens. Our findings also suggest the need for adequate antibiotic treatment in patients with confirmed Aeromonas spp. having enterotoxigenic and/or invasive properties.
AIMS: To study the phenotypic characteristics of Aeromonas spp. from environmental and clinical samples in Spain and to cluster these strains by numerical taxonomy. METHODS AND RESULTS: A collection of 202 Aeromonas strains isolated from bivalve molluscs, water and clinical samples was tested for 64 phenotypic properties; 91% of these isolates were identified at species level. Aeromonas caviae was predominant in bivalve molluscs and Aerom. bestiarum in freshwater samples. Cluster analyses revealed eight different phena: three containing more than one DNA-DNA hybridization group but including strains that belong to the same phenospecies complex (Aerom. hydrophila, Aerom. sobria and Aerom. caviae), Aerom. encheleia, Aerom. trota and three containing unidentified Aeromonas strains isolated from bivalve molluscs. CONCLUSIONS: Aeromonas spp. are widely distributed in environmental and clinical sources. A selection of 16 of the phenotypical tests chosen allowed the identification of most isolates (91%), although some strains remain unidentified, mainly isolates from bivalve molluscs, suggesting the presence of new Aeromonas species. Numerical taxonomy was not in total concordance with the identification of the studied strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Numerical taxonomy of Aeromonas strains isolated from different sources revealed the presence of potentially pathogenic Aeromonas spp., especially in bivalve molluscs, and phena with unidentified strains that suggest new Aeromonas species.
Outbreaks of diarrhea caused by enteropathogens have been reported in day care centers (DCC), but Aeromonas species have not been implicated. This study evaluated 381 children involved in 51 outbreaks in four DCC to determine the association of Aeromonas species with diarrhea and to characterize the isolates. The organism was identified in two outbreaks of diarrhea. In one, Aeromonas species were isolated from 6 (24%) of 25 children and in the other from 5 (21%) of 24 children. Seven other Aeromonas strains from children in DCC were studied. Fourteen (78%) of 18 were Aeromonas caviae and 15 were from children with diarrhea. Of the isolates, 75% did not have plasmids detected; all others had unique plasmid patterns. All strains had different DNA content. Twenty-two control isolates of Aeromonas from children with diarrhea in Mexico and Dallas had different chromosomal DNA patterns. Most Aeromonas infections were associated with symptoms. Chromosomal DNA patterns differentiated Aeromonas strains better than did plasmid DNA patterns. The outbreaks of diarrhea were unusual in that several different Aeromonas genospecies were involved in each outbreak.
During a 20-month period 55 strains of Aeromonas species were isolated from 53 children with diarrhea. The isolation rate of 2.5% for Aeromonas compared with the rates of 4.5% for Shigella, 3.3% for Salmonella, 2.7% for Campylobacter and 0.05% for Yersinia. In 45 children Aeromonas was the sole bacterial enteropathogen identified. Aeromonas was also isolated from 2 (0.5%) of 380 asymptomatic children. Despite its known lack of identifiable virulence properties, Aeromonas caviae was the most prevalent species, accounting for 69% of the isolates. None of the A. caviae strains produced cytotoxin by the 51Cr release assay and 12.5% were weakly enterotoxigenic by the infant mouse assay. All of the Aeromonas sobria and 71% of Aeromonas hydrophila were positive for both toxins. Ninety-two percent of the children with Aeromonas-associated diarrhea were younger than 3 years; 84% of the cases were seen between May and October. The majority of the children had an acute onset of watery diarrhea. Fever and vomiting were most commonly associated with the isolation of A. sobria. Eight children had chronic or intermittent diarrhea lasting for weeks to months before consultation; A. caviae was the isolate in all these cases. Several complications possibly related to Aeromonas intestinal infection were observed. These included Gram-negative bacteremia, intussusception, internal hernia strangulation, hemolytic uremic syndrome and failure to thrive in patients with chronic diarrhea.
We examined the taxonomic position of seven Aeromonas isolates, recovered from Flemish and Scottish drinking water production plants and reservoirs, which were previously recognized by numerical analysis of genomic AFLP fingerprints as members of an unknown Aeromonas taxon that most closely resembled the species Aeromonas bestiarum (DNA hybridization group [HG] 2). The new phenotypic and DNA-DNA hybridization data obtained in this study show that the A. bestiarum-like strains constitute a separate Aeromonas species, for which the name Aeromonas popoffii sp. nov. is being proposed. The new species exhibited an internal DNA relatedness ranging from 79 to 100% and was 22 to 63% related to the type or reference strains of other Aeromonas spp. The highest DNA binding values were determined with A. bestiarum (51 to 63%), followed by Aeromonas hydrophila sensu stricto (HG1; 50 to 60%) and Aeromonas salmonicida (HG3; 39 to 55%). Although fingerprints generated by ribotyping and cellular fatty acid analysis often were highly similar, minor differences between the respective fingerprints were of significance for the differentiation of A. popoffii from its closest taxonomic neighbors, HG1, HG2, and HG3. Phenotypically, all seven strains of A. popoffii were positive for acid and gas production from D-glucose and glycerol, growth in KCN broth, arginine dihydrolase, DNase, Voges-Proskauer reaction, and resistance to vibriostatic agent O/129 and ampicillin but displayed negative reactions for production of urease, tryptophan deaminase, ornithine decarboxylase, and lysine decarboxylase (LDC). None of the strains displayed strong hemolytic activity. The lack of D-sucrose fermentation and LDC production and the ability to utilize DL-lactate as the sole energy and carbon source were useful characteristics for the biochemical separation of A. popoffii from A. bestiarum. Other Aeromonas spp. could be differentiated phenotypically from the new species by at least two features. The chromosomal G+C content of A. popoffii ranges from 57.7 to 59.6 mol%. Strain LMG 17541 is proposed as the type strain.