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Genomic insights into Aeromonas infections in diarrheal patients: high diversity, emerging resistance, and potential outbreak in Beijing.

BACKGROUND: Aeromonas species are ubiquitous aquatic bacteria that have emerged as significant foodborne enteric pathogens worldwide, yet their genomic landscape in clinical settings remains poorly delineated, particularly in Beijing. METHODS: To address this gap, we performed active surveillance for Aeromonas among 691 consecutive diarrheal outpatients in a Beijing district from January to December 2024. Isolates were recovered using enrichment culture coupled with PCR screening and identified by MALDI-TOF MS. Antimicrobial susceptibility was tested against 17 agents. Whole-genome sequencing was conducted on all isolates, enabling average nucleotide identity (ANI) analysis, comprehensive annotation of antimicrobial resistance and virulence genes, multilocus sequence typing (MLST), and core-genome SNP (cgSNP)-based phylogenetics. RESULTS: Aeromonas was detected in 3.3% (23/691) of patients, with Aeromonas veronii (56.52%, 13/23) and Aeromonas caviae (26.09%, 6/23) predominating. Co-infections with other enteric pathogens occurred in 65.2% of positive cases. Resistance rates were notably high for ampicillin/ampicillin-sulbactam (78.26%), nalidixic acid (56.52%), and ertapenem (21.73%), and 65.21% of isolates were multidrug-resistant. Genotypic-phenotypic concordance was robust, with β-lactamase genes ampS (60.87%) and blaCEPH-A3 (47.83%) being most prevalent. Strikingly, mcr-3.25 and mcr-3.3, which belong to the mcr family (originally described as mobile colistin resistance genes), were identified in 8.70% of isolates, exhibiting perfect correlation with phenotypic resistance. Plasmer analysis suggested both mcr genes to be chromosomally encoded. Comparative genomic analysis of virulence-associated genes revealed striking species-specific specialization: A. veronii predominantly carried complete T3SS clusters (61.5%), A. caviae and Aeromonas enteropelogenes were enriched in T6SS genes, and a single A. dhakensis isolate possessed an extensive arsenal including T3SS, T6SS, and a full RTX toxin cluster. MLST resolved the 23 isolates into 22 sequence types, 18 of which were novel. Phylogenetic reconstruction identified a tight monophyletic cluster of three A. veronii isolates (9-27 SNP differences) recovered within a 96-h window, suggestive of a potential cluster that warrants further epidemiological investigation. Comparative genomic analysis of the rare species Aeromonas allosaccharophila demonstrated that the Beijing clinical isolate S14 differs from the U.S. clinical strain ATCC 35942 by 42,099 SNPs, confirming its distinct genetic lineage. CONCLUSION: Collectively, this study delineates high genetic diversity, emerging chromosomal colistin resistance, and species-specific virulence specialization among Aeromonas isolates from diarrheal patients in Beijing. The detection of a potential outbreak cluster and a rare clinical isolate underscores the power of genomics-based surveillance for detecting and mitigating foodborne pathogen threats.

Aeromonas

Snake-to-human transmission of Aeromonas (Pl) shigelloides resulting in gastroenteritis.

A healthy young man developed acute gastroenteritis after handling an infected bao constrictor. The animal died after contracting "mouth-rot disease", a progressive ulcerative stomatitis of snakes charactistically caused a Aeromonas species. Stool cultures from the patient yielded a heavy growth of Aeromonas (Plesiomonas) shigelloides but no other enteric pathogens. Treatment wit sulfamethoxazole-trimethoprim resulted in rapid relief of clinical symptoms. Aeromonas species are not considered part of the normal human fecal flora and gastroenteritis due to this organism is rare. Furthermore, this case appears to represent a new zoonosis: human Aeromonas (Plasiomonas) gastroenteritis derived from contact with an infected animal host.

Acute Disease

An Aeromonas species implicated in ulcer-disease of the cod (Gadus morhua).

Vibrio anguillarum is the most frequently isolated bacterial species involved in ulcer disease in salt water fish. However, an Aeromonas species was very often incriminated in ulcers and septicaemia in cods (Gadus morhua) in Danish coastal areas. These Aeromonas strains were very uniform in their biochemical activities only differing in growth on Simmons citrate agar, and the mean value for the guanine + cytosine content of DNA was 59 per cent with a standard deviation of 0.4. Primary serological investigations demonstrated a possible identity to O and K antigens. The most characteristic biochemical features of these strains were a gas production from glycerol but not from glucose. They were positive in lysine decarboxylase, did not produce indole, and had a typical fermentation pattern of the glycosides. Negative results were found in H2S production, phosphatase and in the utilization of NH4+ and glucose as the only source of N and C. The evident differences from the species described and subspecies of Aeromonas elucidate the weakness of the existing systems of biotyping. A broader conception of the biochemical spectrum for the individual species of Aeromonas combined with serotyping would seem to be a better system for identification, and also for an epidemiological purpose.

Aeromonas

Isolation of enterotoxigenic Aeromonas from fish.

This paper reports two different enterotoxigenic classes of the genus Aeromonas isolated from fishes. Strains of A. sobria were isolated from healthy fishes only wheras strains of A. hydrophila were isolated from both healthy and moribund fishes. Aeromonas sobria and hydrophilia strains produced a cytotoxic factor and were highly proteolytic. Seventy-five percent (75%) of the A. sobria strains produced only one type of hemolysis on 5% blood agar, but 92% of the A. hydrophilia produced two types of hemolysis. Enterotoxigenicity of all strains was tested by the rabbit gut loop technique and the suckling mouse test. Most of those Aeromonads were enterotoxigenic. Enterotoxigenic strains appeared to produce two different enterotoxins which were antigenically related, but they seemed to have different mechanisms of action. Enterotoxins of Aeromonas were shown to be antigenically different from those of E. coli.

Aeromonas

Deoxyribonucleic acid relationships among members of the genus Aeromonas.

Polynucleotide sequences among 24 motile and 11 non-motile aeromonads were studied by analysis of deoxyribonucleic acid - deoxyribonucleic acid (DNA-DNA) duplexes with endonuclease S1. In addition, DNA base composition (mole % guanine and cytosine (G + C)) and relative genome sizes were determined for selected strains. Large variations in genome size were found and % GC ranged from 57.1 to 62.9%. On the basis of the strains examined, the Genus Aeromonas consists of two genotypically legitimate groups: a diverse group of motile aeromonads, and the genetically more homogeneous non-motile aeromonads, comprising the species Aeromonas salmonicida. Internal homology groups could not be demonstrated within the motile aeromonads, and significant divergence in related sequences was indicated. This diverse motile group forms the single species Aeromonas hydrophila.

Aeromonas

Human aeromonas infections: a review of the literature and a case report of endocarditis.

Our patient, with cirrhosis and chronic renal failure, represents an example of the susceptibility of a compromised host to Aeromonas infections. This patient, however, differs from previously reported cases in at least two important aspects. First, it is possible that her portal of entry was a fresh A-V fistula puncture site rather than an intestinal site. The temporal relationship of exposure to flood water prior to the onset of sepsis lends support to this possibility. Epidemiologic investigation of the dialysis center failed to reveal Aeromonas isolates from cultures of the water supply, machinery, or other patients. Second, this case is unique in that our patient developed a destructive aortic valve endocarditis resulting in valvular perforations and acute aortic insufficiency. Furthermore, this infection was initiated on what appears to have been a previously normal valve. Based on a review of the literature and the virulence demonstrated by A. hydrophila in our patient, we conclude that organisms of the genus Aeromonas are capable of inducing serious human infection. Such infections are more likely to occur in compromised hosts. A. hydrophila has accounted for the majority of reported infections.

Abdomen

[Use of the A-2 elective medium for the differentiation of Aeromonas from related microorganisms].

The trial of culture medium A-2, proposed for the identification of Aeromonas on the basis of the nutritional utilization of gelatin as the only source of nitrogen and starch as the source of carbon in view of the limited content of mineral components in the medium, revealed that no other species of the family Vibronaceae grew in this medium, and it could be used in an additional test for the differentiation of vibrios from Aeromonas. A group of strains isolated from human feces, surface water and sewage was found to be incapable of growing in culture medium A-2; these strains, though having a number of characteristics in common with Aeromonas, differed from them in a number of additional tests.

Aeromonas

Aeromonas hydrophila sepsis in a patient undergoing hemodialysis therapy.

Aeromonas hydrophila septicemia complicated by a generalized cutaneous vasculitis developed in a patient receiving home hemodialysis therapy. Because the Aeromonas organism is found in many natural water sources, the possibility that this patient's hemodialysis system became contaminated was explored. Although cultures from the patient's home environment showed no Aeromonas sp, the possibility still exists that the site of contamination was in the dialysis system.

Aeromonas

Proteomics-based analysis of the defense mechanisms of disease-resistant grass carp against Aeromonas veronii.

Sustainable aquaculture of grass carp (Ctenopharyngodon idella, GC) is consistently threatened by bacterial diseases, particularly those caused by Aeromonas veronii. A disease-resistant grass carp (DR-GC) has been developed by backcrossing female gynogenetic GC with normal male GC, exhibiting improved resistance. However, the systemic molecular mechanisms of DR-GC defending against Aeromonas veronii infection remain largely unexplored. Here, a label-free quantitative proteomics approach was employed to systematically compare proteomic profiles across five tissues (intestine, liver, muscle, skin, and kidney) in DR-GC and GC under healthy and infected conditions. The intestine was identified as the central defense tissue, exhibiting the highest number of differentially abundant proteins (DAPs). In DR-GC, A0A3N0YEK7 (small ribosomal subunit protein eS28), A0A3N0YGT8 (ATP synthase-coupling factor 6) and A0A3N0YNS7 (apolipoprotein A-I) were significantly upregulated in intestine, while D5KZW6 (GCHV-induced protein), A0A3N0Z0A1 and Q8JH84 (hemoglobin subunit alpha) were significantly dysregulated across multiple tissues, which playing the critical roles in defense mechanisms at the protein level. Furthermore, cytochrome P450-associated pathways, cytosolic DNA-sensing and RIG-I-like receptor signaling pathways were identified as crucial coordinators mediating immune and metabolic responses. This study provides the first comprehensive proteomic view of multi-tissue defense mechanisms in DR-GC, and identifies key DAPs and pathways for subsequent functional validation.

Animals

A species-discriminatory aerA TaqMan qPCR assay for rapid quantification of Aeromonas veronii in fish tissues and aquaculture water.

Aeromonas veronii is a major bacterial pathogen in freshwater aquaculture, yet rapid species-level quantification remains challenging within the genetically complex genus Aeromonas. We developed a singleplex hydrolysis-probe (TaqMan) quantitative PCR (qPCR) assay targeting an A. veronii-discriminatory region of the aerolysin gene (aerA) and validated it according to MIQE recommendations. Plasmid standards gave a linear range of 2 to 2 × 106 copies/reaction (R2 = 0.9962) with 100.5% amplification efficiency. The endpoint limit of detection was 2 copies per reaction, and 20 copies per reaction was set as the practical reporting limit based on reproducible detection and low intra- and inter-assay variation. Analytical specificity was evaluated with genomic DNA from an 18-strain panel, with reproducible amplification observed only for A. veronii. The assay was further tested in 55 fish-tissue and 11 aquaculture-water DNA extracts. NH8B-1D2 sample-process monitoring was used for matrix-level recovery correction, and tissue and water extraction blanks were undetermined. The aerA target was detected in all tested gill, stomach/intestine, spleen, kidney/head kidney, pond-water filter and Xiamen seawater filter extracts, and in 10/11 liver extracts. Median NH8B-corrected loads were highest in gill among tissues and higher in pond-water filters than in Xiamen seawater filters. A separate Vibrio harveyi inhibition-check assay indicated no obvious amplification-stage inhibition. This assay supports rapid quantification of aerA-positive A. veronii in fish and aquaculture-water matrices.

Animals

Aeromonas primary wound infection of a diver in polluted waters.

Two separate species of Aeromonas, A. sobria (not listed as a species in Bergey's Manual of Determinative Bacteriology, 8th ed.) and A. hydrophila, were primary pathogens isolated from the leg wound of a diver conducting operations in polluted waters. This is the first recorded instance of a primary infection of soft tissue in a human caused by two species of Aeromonas, one of which was resistant to tetracycline. Because of the very rapid development of this wound infection, cytotoxicity of these organisms was examined in several biological systems. A. sobria was hemolytic for sheep erythrocytes, cytotoxic for Y-1 adrenal cells, and enterotoxic in rabbit ligated intestinal loops, whereas A. hydrophila was hemolytic and cytotoxic. Pertinent clinical, bacteriological, and environmental features of the case are presented.

Adult

[Antigenic relations between Vibrio cholerae and Aeromonas hydrophila].

The antigenic relationships between Vibrio cholerae and the oxidase-positive intestinal bacteria, agglutinated on slides by cholera antiserum, were investigated by the agglutination reaction. Of the 478 oxidase-positive strains studied only strain 209 A, belonging to species Aeromonas hydrophila was selected. Evidence was found of common agglutinogenic fractions between Vibrio cholerae and Aeromonas hydrophila 209 A. Attention is drawn to possible confusions in the preliminary stages of a laboratory diagnosis of cholera.

Aeromonas

[Aeromonas punctata subsp. caviae as the causative agent of acute gastroenteritis (author's transl)].

Since in the past, Aeromonas hydrophila had been isolated from all cases of human infection described. A. punctata and the anaerogenic sub-species were considered as apathogenic. From the case described, a close association between acute diarrhea with vomiting and the identification of A. punctata subsp. caviae becomes evident so that a conditional pathogenicity of this sub-species must be assumed. The question is discussed whether a preceding disturbance of the intestinal habitat in the presence of a particular susceptibility of the gastrointestinal tract promoted gastro-enteritis. Infection may have been brought about by the ingestion of surface water contaminated by sewage. Attention is again drawn to the fact that in the case of enteritis occuring during the open-air bathing season, faeces samples should also be examined for their Aeromonas content which may be performed by a simple indophenol or so-called cytochrome oxidase reaction of the aerobic flora in feces by rubbing off colonies into a corresponding test strip.

Acute Disease

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

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

Liver transcriptome analysis revealed multiple immune processes and lipid metabolism pathways involved in the defense response of the turbot (Scophthalmus maximus) against Aeromonas salmonicida.

Aeromonas salmonicida is a significant pathogen causing notable economic losses in Scophthalmus maximus aquaculture. This study utilized Illumina sequencing technology to examine the transcriptional response characteristics of S. maximus liver at 24 h following A. salmonicida infection. A total of 2363 differentially expressed genes (DEGs) were identified when compared to the negative control group. The immunity-related Toll-like receptor signaling pathway, NOD-like receptor signaling pathway, as well as metabolism-related PPAR signaling pathway and insulin signaling pathway, were notably enriched. Significant differences exist in the expression of key genes within the PPAR pathway, particularly cd36, acsl4a, pparαa, and plin2, all of which mediate the interaction between lipid metabolism and the immune response. These results offer valuable insights into the immunometabolic regulatory mechanism of S. maximus response to A. salmonicida infection.

Animals

Genome-wide characterization of NOD-like receptor genes links NLR repertoire evolution to spleen immune responses after Aeromonas hydrophila challenge in the Chinese spiny frog (Quasipaa spinosa).

NOD-like receptors (NLRs) are cytosolic pattern-recognition receptors that detect pathogen-associated and damage-associated molecular patterns and mediate innate immune signaling in vertebrates. However, the genomic repertoire, evolutionary diversification, and infection-associated expression of NLR genes remain poorly defined in non-model amphibians. In this study, 66 NLR genes were identified from the Chinese spiny frog (Quasipaa spinosa) genome and designated as QsNLR1-QsNLR66. These genes were unevenly distributed across chromosomes and were classified into three phylogenetic groups, with most members exhibiting conserved motif architectures. Gene duplication analysis indicated that dispersed duplication was the main contributor to QsNLR expansion. Synteny analysis detected five conserved orthologous gene pairs between Q. spinosa and Pelophylax nigromaculatus, suggesting partial conservation of NLR genomic organization between the two amphibians. Ka/Ks analysis showed that several duplicated gene pairs, including NLRC3-like/QsNLR36 and NLRC3-like/QsNLR50, exhibited Ka/Ks ratios greater than one, suggesting potential sequence divergence after duplication. Spleen RNA sequencing (RNA-seq) after Aeromonas hydrophila challenge revealed enrichment of immune-related Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Weighted gene co-expression network analysis linked several QsNLRs to infection-associated modules, among which QsNLR57 was co-expressed with CYBB, ADAM17, SPI1, and HK2. RT-qPCR using time-matched phosphate-buffered saline (PBS) controls showed distinct temporal patterns, with stronger induction of QsNLR29, QsNLR57, and QsNLR66 and weaker or delayed responses of QsNLR50 and QsNLR56. These results characterize the NLR repertoire of Q. spinosa and identify infection-associated QsNLR candidates for future studies of antibacterial immunity in amphibians.

Animals