Phenotypic and genotypic analysis of antimicrobial resistance and population structure of gastroenteritis-related Aeromonas isolates.

BACKGROUND: The population structure and the correlation between antimicrobial resistance (AMR) phenotypes and genotypes in Aeromonas species isolated from patients with gastroenteritis are not well understood. The aims of the study were to: (1) investigate the antimicrobial susceptibility profiles of Aeromonas species isolated from patients with gastroenteritis; (2) explore the relationship between AMR genes and resistance phenotypes; and (3) describe the population structure of these isolates and provide evidence of transmission events among them. METHODS: This microbiological survey was performed at the Microbiology Laboratory of the Emek Medical Center in Afula, Israel. Cultivation of Aeromonas was attempted from stool samples that tested positive by PCR. Antimicrobial susceptibility testing (AST) was performed using the Sensititre GN3F microdilution panel. Whole genome sequencing (WGS) was done using the Illumina NextSeq500/550 system. Phylogenetic studies involved multi-locus sequence typing (MLST) and core genome (cg) MLST. Resistance mechanisms were identified using the Comprehensive Antibiotic Resistance Database and compared with the AST results. RESULTS: The study included 67 patient-unique isolates. The species that were identified included A. caviae (n = 58), A. dhakensis (n = 3), A. media (n = 2), A. veronii (n = 2) and A. hydrophila (n = 2). Isolates were almost uniformly susceptible to amikacin, gentamicin, aztreonam, cefepime, ceftazidime, ciprofloxacin and meropenem. All isolates with the exception of 1-2 isolates were resistant to ampicillin, cefazolin and ampicillin-sulbactam which was compatible with the presence of the blaOXA genes. Variable resistance rates were observed to cefuroxime, cefoxitin, ceftriaxone, piperacillin-tazobactam that were not correlated with the presence of other ß-lactamase genes. Resistance to tetracycline and trimethoprim-sulfamethoxazole correlated with the presence of tetA and sul1, respectively. The population structure of A. caviae was highly diverse with the minority of the isolates (16/57) clustering into six defined sequence types. A cgMLST-based distance of four genes was found in one pair of isolates, suggesting common source transmission. CONCLUSIONS: A. caviae is the dominant species related to gastroenteritis and is characterized by a diverse population structure, with almost no evidence for common-source transmission. Resistance rates to most antimicrobial agents were low and partially matched with the presence of resistance genes.

Molecular detection of enterotoxins in multidrug resistant Aeromonas from ready to eat foods in North Western Himalayas: Public health significance.

Aeromonas spp. are normal inhabitants of aquatic environments and are emerging foodborne bacterial pathogens. Aeromonas spp. contamination is frequent in ready-to-eat (RTE) seafood and can also occur in products prepared from milk or meat. The study determined the enterotoxin and antimicrobial susceptibility profiles of Aeromonas spp. isolates recovered from RTE milk products (n = 105), RTE meat/fish products (n = 40) and drinking water (n = 60) samples collected from tourist places in Himachal Pradesh, India, in northwestern Himalayas. 7.3 % (16/220) samples were found contaminated with Aeromonas spp. These isolates were identified as A. hydrophila (31.3 %), A. schubertii (25.0 %), A. sobria (25.0 %) and A. veronii (18.8 %). Aeromonas spp. contamination was significantly higher (14.3 %, 15/105, p = 0.0001) in RTE milk products. The contamination levels for water samples were 1.7 % whereas none of the tested RTE meat or fish products yielded Aeromonas spp. Among RTE milk products, contamination was significantly higher in paneer (South Asian soft cheese) (26.1 %, p = 0.0027) and cream (25.0 %, p = 0.046) based RTE foods. All isolates carried alt (361 bp), encoding a cytotonic heat-labile enterotoxin. Ampicillin resistance was 100 % and high levels (>30 %) of resistance were recorded for amoxicillin/clavulanic acid, amikacin, cefotaxime and ceftazidime. Six (37.5 %) isolates were multi drug resistant (MDR), showing resistance to aminoglycosides, cephams and penicillins. Isolation of alt carrying MDR isolates from RTE foods indicates that Aeromonas spp. can be potential foodborne public health threat in northwestern Himalayas.

Genomic characterization of Aeromonas spp. isolates from striped catfish with motile Aeromonas septicemia and human bloodstream infections in Vietnam.

Aeromonas spp. are commonly found in the aquatic environment and have been responsible for motile Aeromonas septicemia (MAS) in striped catfish, resulting in significant economic loss. These organisms also cause a range of opportunistic infections in humans with compromised immune systems. Here, we conducted a genomic investigation of 87 Aeromonas isolates derived from diseased catfish, healthy catfish and environmental water in catfish farms affected by MAS outbreaks in eight provinces in Mekong Delta (years: 2012-2022), together with 25 isolates from humans with bloodstream infections (years: 2010-2020). Genomics-based typing method precisely delineated Aeromonas species while traditional methods such as aerA PCR and MALDI-TOF were unable identify A. dhakensis. A. dhakensis was found to be more prevalent than A. hydrophila in both diseased catfish and human infections. A. dhakensis sequence type (ST) 656 followed by A. hydrophila ST251 were the predominant virulent species-lineages in diseased catfish (43.7 and 20.7 %, respectively), while diverse STs were found in humans with bloodstream infections. There was evidence of widespread transmission of ST656 and ST251 on striped catfish in the Mekong Delta region. ST656 and ST251 isolates carried a significantly higher number of acquired antimicrobial resistance (AMR) genes and virulence factors in comparison to other STs. They, however, exhibited several distinctions in key virulence factors (i.e. lack of type IV pili and enterotoxin ast in A. dhakensis), AMR genes (i.e. presence of imiH carbapenemase in A. dhakensis), and accessory gene content. To uncover potential conserved proteins of Aeromonas spp. for vaccine development, pangenome analysis has unveiled 2202 core genes between ST656 and ST251, of which 78 proteins were in either outer membrane or extracellular proteins. Our study represents one of the first genomic investigations of the species distribution, genetic landscape, and epidemiology of Aeromonas in diseased catfish and human infections in Vietnam. The emergence of antimicrobial resistant and virulent A. dhakensis strains underscores the needs of enhanced genomic surveillance and strengthening vaccine research and development in preventing Aeromonas diseases in catfish and humans, and the search for potential vaccine candidates could focus on Aeromonas core genes encoded for membrane and secreted proteins.

Novel Qnr Families as Conserved and Intrinsic Quinolone Resistance Determinants in Aeromonas spp.

The environment has been identified as an origin, reservoir, and transmission route of antibiotic resistance genes (ARGs). Among diverse environments, freshwater environments have been recognized as pivotal in the transmission of ARGs between opportunistic pathogens and autochthonous bacteria such as Aeromonas spp. In this study, five environmental strains of Aeromonas spp. exhibiting multidrug resistance (MDR) were selected for whole-genome sequencing to ascertain their taxonomic assignment at the species-level and to delineate their ARG repertoires. Analyses of their genomes revealed the presence of one protein almost identical to AhQnr (A. hydrophila Qnr protein) and four novel proteins similar to AhQnr. To scrutinize the classification and taxonomic distribution of these proteins, all Aeromonas genomes deposited in the NCBI RefSeq genome database (1,222 genomes) were investigated. This revealed that these Aeromonas Qnr (AQnr) proteins are conserved intrinsic resistance determinants of the genus, exhibiting species-specific diversity. Additionally, structure prediction and analysis of contribution to quinolone resistance by AQnr proteins of the isolates, confirmed their functionality as quinolone resistance determinants. Given the origin of mobile qnr genes from aquatic bacteria and the crucial role of Aeromonas spp. in ARG dissemination in aquatic environments, a thorough understanding and strict surveillance of AQnr families prior to the clinical emergence are imperative. In this study, using comparative genome analyses and functional characterization of AQnr proteins in the genus Aeromonas, novel Aeromonas ARGs requiring surveillance has suggested.

Description of a pathogenic strain of Aeromonas dhakensis isolated from Ancherythroculter nigrocauda in an inland region of China.

Aeromonas dhakensis is reported as an emerging pathogenic species within the genus Aeromonas and is widely distributed in tropical coastal areas. This study provided a detailed description and characterization of a strain of A. dhakensis (202108B1) isolated from diseased Ancherythroculter nigrocauda in an inland region of China. Biochemical tests identified the isolate at the genus level, and the further molecular analysis of concatenated housekeeping gene sequences revealed that the strain belonged to the species A. dhakensis. The isolated A. dhakensis strain was resistant to five antibiotics, namely, penicillin, ampicillin, clindamycin, cephalexin, and imipenem, while it was susceptible to or showed intermediate resistance to most of the other 15 tested antibiotics. The isolated strain of A. dhakensis caused acute hemorrhagic septicemia and tissue damage in artificially infected A. nigrocauda, with a median lethal dose of 7.76 × 104 CFU/fish. The genome size of strain 202108B1 was 5 043 286 bp, including 1 chromosome and 4 plasmids. This is the first detailed report of the occurrence of infection caused by an A. dhakensis strain causing infection in an aquaculture system in inland China, providing important epidemiological data on this potential pathogenic species.

AsaGEI2d: a new variant of a genomic island identified in a group of Aeromonas salmonicida subsp. salmonicida isolated from France, which bears the pAsa7 plasmid.

Genomic islands (Aeromonas salmonicida genomic islands, AsaGEIs) are found worldwide in many isolates of Aeromonas salmonicida subsp. salmonicida, a fish pathogen. To date, five variants of AsaGEI (1a, 1b, 2a, 2b and 2c) have been described. Here, we investigate a sixth AsaGEI, which was identified in France between 2016 and 2019 in 20 A. salmonicida subsp. salmonicida isolates recovered from sick salmon all at the same location. This new AsaGEI shares the same insertion site in the chromosome as the other AsaGEI2s as they all have a homologous integrase gene. This new AsaGEI was thus named AsaGEI2d, and has five unique genes compared to the other AsaGEIs. The isolates carrying AsaGEI2d also bear the plasmid pAsa7, which was initially found in an isolate from Switzerland. This plasmid provides resistance to chloramphenicol thanks to a cat gene. This study reveals more about the diversity of the AsaGEIs.

Taxonomy, virulence determinants and antimicrobial susceptibility of Aeromonas spp. isolated from bacteremia in southeastern China.

BACKGROUND: The study aimed to elucidate the species taxonomy, clinical manifestations, virulence gene profiles and antimicrobial susceptibilities of Aeromonas strains isolated from life-threatening bacteremia in southeastern China. METHODS: Clinical samples of Aeromonas causing bacteremia were isolated from a teaching hospital in Wenzhou from 2013 to 2018 and a retrospective cohort study was performed. Aeromonas strains were identified at species level by housekeeping gene gyrB. Virulence and drug resistance-associated genes were screened by polymerase chain reaction (PCR) and antimicrobial susceptibility testing (AST) was performed by the VITEK 2 Compact system. RESULTS: A total of 58 Aeromonas isolated from patients with bacteremia were collected during 6 years (2013-2018). 58 isolates were identified to five different species, where Aeromonas dhakensis appeared to be the predominant species (26/58), followed by Aeromonas veronii (13/58), Aeromonas caviae (10/58), Aeromonas hydrophila (7/58) and Aeromonas jandaei (2/58). 16 of 58 patients had poor prognosis. Poor prognosis was significantly associated with liver cirrhosis and inappropriate empirical antimicrobials therapy. The progression of bacteremia caused by Aeromonas was extremely fast, especially in A. dhakensis infections. Virulence genes aer, lip, hlyA, alt, ast, and act, were detected at ratios of 24.1% (14/58), 62.1% (36/58), 65.5% (38/58), 58.6% (34/58), 15.5% (9/58) and 65.5% (38/58), respectively. Antimicrobial susceptibility testing exhibited that 9 out of 58 isolates were identified as multi-drug resistant (MDR) organism. The blaTEM gene was identified in all 9 MDR isolates. blaSHV, blaAQU-1, blaMOX, blaCepH, blaCphA and aac(6')-Ib-cr were detected in 4 isolates, 2 isolates, 1 isolate, 3 isolates, 8 isolates, and 3 isolates, respectively. The majority of Aeromonas strains maintained susceptible to 3rd generation cephalosporins, aminoglycosides, fluoroquinolones and furantoin. CONCLUSIONS: The prevalence and dangerousness of Aeromonas infections, especially A. dhakensis, are underestimated in clinic. Continuous monitoring is essential to keep track of MDR Aeromonas due to the increasing prevalence recently and a more effective measure is required to control the spread of resistance determinants.

Antimicrobial resistance in Aeromonas species isolated from aquatic environments in Brazil.

AIM: The current study was conducted to determine the antimicrobial resistance profile and genetic relatedness of Aeromonas sp. isolated from healthcare and urban effluents, wastewater treatment plant (WWTP) and river water. METHODS AND RESULTS: We detected the presence of genes conferring resistance to ß-lactam, quinolone and aminoglycoside. Multilocus sequence typing was carried out to differentiate the strains, and multilocus phylogenetic analysis was used to identify the species. A total of 28 cefotaxime-resistant Aeromonas sp. strains were identified, harbouring uncommon Guiana-extended-spectrum (GES)-type ß-lactamases (GES-1, GES-5, GES-7 and GES-16). Multidrug-resistant Aeromonas sp. were found in hospital wastewater, WWTP and sanitary effluent, and A. caviae was identified as the most prevalent species (85·7%). CONCLUSION: The release of untreated healthcare effluents, presence of antimicrobials in the environment, in addition to multidrug-resistant Aeromonas sp., are all potential factors for the spread of resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: We identified a vast repertoire of antimicrobial resistance genes (ARG) in Aeromonas sp. from diverse aquatic ecosystems, including those that encode enzymes degrading broad-spectrum antimicrobials widely used to treat healthcare-associated infections. Hospital and sanitary effluents serve as potential sources of bacteria harbouring ARG and are a threat to public health.

Aeromonas spp. isolated from ready-to-eat seafood on the Norwegian market: prevalence, putative virulence factors and antimicrobial resistance.

AIMS: We aim to investigate the prevalence, putative virulence factors and antimicrobial resistance of mesophilic Aeromonas isolated from ready-to-eat (RTE) seafood available on the Norwegian market, and to assess the potential risks by consuming RTE seafood to consumers. METHODS AND RESULTS: The prevalence of mesophilic Aeromonas in 148 RTE seafood was investigated and the highest prevalence was found in retail sushi (17%), followed by oysters (10%), fresh salmon loins (10%) and scallops (4%). Among 43 Aeromonas isolates, 75% of them were identified as A. media, 23% as A. salmonicida and 2% as A. bestiarum based on partial gryB gene sequencing. Aeromonas isolates were potentially pathogenic due to the presence of four virulence genes: alt (73%), hylA (22%), aerA (17%) and act (6%). In addition, all isolates were resistant to ampicillin and erythromycin. Most of the isolates (98%) were multidrug resistant. CONCLUSIONS: The occurrence of potentially pathogenic and multidrug-resistant Aeromonas strains in RTE seafood implies a potential risk to consumers. Our finding suggests that RTE seafood could be a potential vehicle for the transfer of virulent and multidrug-resistant Aeromonas. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first study to report multiple antibiotic resistance in Aeromonas associated with RTE seafood in Norway.

Isolation and molecular identification of Aeromonas species from the tank water of ornamental fishes.

Aeromonads are recognised as important pathogens of fishes. In this study, ten water samples were randomly collected from pet shops' fish tanks and home aquaria inhabited by several fish species (silver arowana, koi, goldfish, catfish, pictus fish, silver shark and silver dollar fish). Altogether 298 colonies were isolated using Aeromonas selective agar. A total of 154 isolates were then confirmed as belonging to the genus Aeromonas using the GCAT gene. Using ERIC-PCR, a total of 40 duplicate isolates were excluded from the study and 114 isolates were subjected to PCR-RFLP targeting the RNA polymerase sigma factor (rpoD) gene using lab-on-chip. A total of 13 different Aeromonas species were identified. The most prevalent species were A. veronii (27%, 31/114), followed by A. dhakensis (17%, 19/114), A. finlandiensis (9%, 10/114), A. caviae (8%, 9/114), A. hydrophila (4%, 4/114), A. jandaei (4%, 4/114), A. rivuli (3%, 3/114), A. enteropelogens (2%, 2/114), A. tecta (2%, 2/114), A. allosaccharophila (1%, 1/114), A. eucrenophila (1%, 1/114), A. media (1%, 1/114) and A. diversa (1%, 1/114). Twenty-six isolates (23%) were unidentifiable at species level. The present study demonstrates that Aeromonas species are highly diverse in freshwater fish tanks, and suggests the potential risks posed by the isolated bacteria to the health of ornamental fish species.

Occurrence, diversity and temperature-dependent growth kinetics of Aeromonas spp. in lettuce.

Bagged, pre-cut and prewashed lettuce products are marketed as ready to eat. This concept poses a food safety concern, due to lack of efficient hurdles to eliminate possible microbial contaminants from the fresh produce and/or the processing itself. Aeromonas spp. are potential foodborne pathogens that are frequently isolated from lettuce. High counts of, e.g., A. hydrophila have been found in retail ready-to-eat (RTE) vegetable salads. The aim of this study was to assess the general microbiological quality, the occurrence and diversity of potential human pathogenic mesophilic Aeromonas spp. of retail RTE lettuce products. Additionally, temperature-dependent growth kinetic parameters of Aerobic Plate Counts (APC) and Aeromonas spp. in one selected RTE lettuce product, rocket lettuce, were quantified by performing storage experiments at 4 °C, 8 °C and 12 °C. The Aeromonas isolates were further characterized regarding pathogenic traits and phylogenetic relationship. The overall hygienic quality of the lettuce products was unsatisfactory, as 60% of the products had an APC level higher than 7.0 log CFU/g. Presumptive Aeromonas spp. were detected in 52% of the samples, levels ranging from approximately 2.0-6.0 log CFU/g. Significantly lower counts of APC and Aeromonas spp. were found in uncut and unwashed products. Presumptive Aeromonas spp. were able to proliferate in rocket lettuce stored at 4 °C (µmax = 0.39 ± 0.06/d and µmax = 0.43 ± 0.05/d for lettuce from producers A and B, respectively), and µmax was approximately 2× higher at 8 °C and 3× higher at 12 °C. Eighty-four percent of the collected isolates were identified as A. media, based on partial gyrB sequencing. Additionally A. salmonicida and A. bestiarum were detected. The pathogenic potential in this material was high, most of the isolates harbored at least one of the toxin genes, act, ast, alt.

Rapid Fulminant Progression and Mortality Secondary to Aeromonas dhakensis Septicemia with Hepatitis B Virus Infection Following the Ingestion of Snakehead Fish in Mainland China: A Case Report.

Aeromonas dhakensis is an important ubiquitous Gram-negative and freshwater bacterium detected in different reservoirs. It can cause invasive diseases in humans. Herein, we report the first case in Mainland China of a fulminant death of a 29-year-old man as a result of a new, unexpected association between septicemic A. dhakensis and hepatitis B viral infection (HBV). Herein, the patient died from multiple organ failure 5 d postadmission after the ingestion of Snakehead Fish meal. The isolated bacterium was initially misidentified as Aeromonas hydrophila using VITEK-2, while whole-genome sequencing (WGS) revealed that the isolate is A. dhakensis. WGS revealed the occurrence of three antimicrobial genes of resistance: imiH, cphA2, and blaOXA-12; besides, major virulence factors were detected. In silico, multilocus sequence typing (MLST) showed that our A. dhakensis 17FW001 belonged to a novel sequence type (ST557). A comparative genomic analysis of our isolate with nine selected Aeromonas species was done, which elucidated the pathogenicity of our A. dhakensis. In conclusion, we reported for the first time the association between A. dhakensis and HBV in Mainland China. We revealed that septicemic A. dhakensis could result in severe adverse clinical outcomes that end up with unexpected fulminant death especially when it is accompanied with HBV and sheds light on the virulence of A. dhakensis and the high rate of its misdiagnosis that requires to urgently consider screening of all cases of A. dhakensis for HBV in the future. Besides, caution should be taken while dealing with snakeheads which act as a vector for A. dhakensis.

Pathogenic characterization of Aeromonas salmonicida subsp. masoucida turbot isolate from China.

Aeromonas salmonicida is a gram-negative bacterium that is the causative agent of furunculosis. An A. salmonicida strain was isolated from diseased turbot (Scophthalmus maximus) with the sign of furunculosis from North China. Based on vapA gene, the strain was further classified as A. salmonicida subsp. masoucida RZ6S-1. Culturing RZ6S-1 strain at high temperature (28°C) obtained the virulence attenuated strain RZ6S. Genome sequence comparison between the two strains revealed the loss of the type IV secretion system (T4SS) and type III secretion system (T3SS) from the native plasmid pAsmB-1 and pAsmC-1 of wild-type strain RZ6S-1, respectively. Further study demonstrated that the wild-type strain RZ6S-1, but not its derivative mutant RZ6S, can stimulate apoptosis. Elevated protein level of cleaved caspase-3 was detected from epithelioma papulosum cyprinid (EPC) cells infected with wild-type strain RZ6S-1 as compared with that infected with RZ6S strain. Meanwhile, the invasion of the mutant strain RZ6S was about 17-fold higher than the wild-type strain RZ6S-1, suggesting that some protein(s) from A. salmonicida subsp. masoucida RZ6S-1 suppress its invasion. The RZ6S mutant strain was attenuated, since its LD50 is over 10,000 times higher compared to the wild-type strain as revealed in the turbot infection model.

Aeromonas spp. Isolated from Pacific Abalone (Haliotis discus hannai) Marketed in Korea: Antimicrobial and Heavy-Metal Resistance Properties.

Antimicrobial and heavy-metal resistance of 29 Aeromonas spp. (Aeromonas hydrophila n = 9, Aeromonas enteropelogenes n = 14, Aeromonas veronii n = 3, Aeromonas salmonicida n = 2, and Aeromonas sobria n = 1) isolated from Pacific abalone marketed in Korea were analyzed. All isolates were found to be resistant against ampicillin. High level of resistant to cephalothin (86%), rifampicin (73%), imipenem (42%), and oxytetracycline (35%) were also detected. Thirteen (45%) of the isolates showed multiple antimicrobial resistance (MAR) index ≥ 0.2. The PCR assays implied the presence of qnrS, qnrB, qnrA, tetB, tetA, aac (3')- IIa, aac(6')-Ib, aphAI-IAB, blaCTX, blaTEM, and intI1 genes among 76%, 28%, 14%, 17%, 3%, 3%, 41%, 10%, 41%, 28%, and 66% of the isolates, respectively. Class 1 integron gene cassette profiles aadA1(3%) and aadA2 (3%) were also identified. Lead (Pb) resistance was the highest (69%) among 5 heavy metals tested, whereas 38%, 27%, and 20% of the isolates were resistant to Cadmium (Cd), Chromium (Cr), and Copper (Cu), respectively. Heavy-metal resistance genes, CopA, CzcA, and merA were positive in 83%, 75%, and 41% of the isolates, respectively. In conclusion, observed genotypic and phenotypic resistance profiles of Aeromonas spp. against antimicrobials and heavy metals reveal the ability of serving as a source of antimicrobials and heavy-metal-resistant traits.

Genetic relatedness and novel sequence types of clinical Aeromonas dhakensis from Malaysia.

Aeromonas dhakensis is an emergent human pathogen with medical importance. This study was aimed to determine the sequence types (STs), genetic diversity, and phylogenetic relationships of different clinical sources of 47 A. dhakensis from Malaysia using multilocus sequence typing (MLST), goeBURST, and phylogenetic analyses. The analysis of a concatenated six-gene tree with a nucleotide length of 2994 bp based on six housekeeping genes (gyrB, groL, gltA, metG, ppsA, and recA) and independent analyses of single gene fragments was performed. MLST was able to group 47 A. dhakensis from our collection into 36 STs in which 34 STs are novel STs. The most abundant ST521 consisted of five strains from peritoneal fluid and two strains from stools. Comparison of 62 global A. dhakensis was carried out via goeBURST; 94.4% (34/36) of the identified STs are novel and unique in Malaysia. Two STs (111 and 541) were grouped into clonal complexes among our strains and 32 STs occurred as singletons. Single-gene phylogenetic trees showed varying topologies; groL and rpoD grouped all A. dhakensis into a tight-cluster with bootstrap values of 100% and 99%, respectively. A poor phylogenetic resolution encountered in single-gene analyses was buffered by the multilocus phylogenetic tree that offered high discriminatory power (bootstrap value = 100%) in resolving all A. dhakensis from A. hydrophila and delineating the relationship among other taxa. Genetic diversity analysis showed groL as the most conserved gene and ppsA as the most variable gene. This study revealed novel STs and high genetic diversity among clinical A. dhakensis from Malaysia.

Correction of the type strain of Aeromonas punctata (Zimmermann 1890) Snieszko 1957 and of A. punctata subsp. punctata from ATCC 15468T to NCMB 74T (=NCIMB 74T= ATCC 23309T).

Under Rule 23a (Note 4) of the Bacteriological Code we ask the Judicial Commission to issue an opinion that will correct two errors that were made on the original 1980 Approved Lists of Bacterial Names. We request that the type strain designations for Aeromonas punctata and Aeromonas punctata subsp. punctata be corrected from ATCC 15468T to NCMB 74T. We also ask that the opinion state the 'correct' or best way to write the author citations for several other Aeromonas names in order to avoid future instability in nomenclature when the citations are given.

Conservation of the name Aeromonas eucrenophila over the name Aeromonas punctata for the organism based on type stain NCMB 74T and universally defined as 'Aeromonas DNA hybridization group 6'.

In a companion paper, we requested the Judicial Commission to correct the type strain of Aeromonas punctata from ATCC 15468T to NCMB 74T (=ATCC 23309T). Correction of this error on the 1980 Approved Lists by an Opinion of the Judicial Commission will remove the status of the name Aeromonas caviae as a junior objective synonym of A. punctata. This is important because the scientific community continues to use the name A. caviae almost exclusively instead of A. punctata. However, the corrective action of this Opinion will cause a new problem. A. punctata and A. eucrenophila will then become objective synonyms because both species will have the same type strain NCMB 74T, and A. punctata would have priority because it was published first (1890 vs. 1987). Thus, A. punctata rather than A. eucrenophila would become the correct name for DNA hybridization group 6. A. punctata has had a very confusing history since it was first described as Bacillus punctatus by Zimmermann in 1890. It was without a type strain for over 50 years, and unfortunately, has had an incorrect type strain for some 40 years. The name A. punctata as a bacterial species has been used incorrectly in the literature very frequently, either based on the wrong type strain or with the wrong definition or circumscription. The name A. punctata is not accepted or used by most specialists who study and publish scientific papers and reviews on Aeromonas. Under the heading 'Rejection of Names' Rule 56a of the Bacterial Code states reasons why the Judicial Commission can reject a name, the first is: '(1) An ambiguous name (nomen ambiguum), i.e., a name which has been used with different meanings and thus has become a source of error'. Rule 56a gives the Judicial Commission authority to place names on the list of rejected names. Our analysis of its history leads us to state unequivocally that A. punctata currently is, and has been throughout the vast majority of its history, an ambiguous name. After considering all the possible alternatives and their consequences we request the Judicial Commission to go against the rules of priority; to invoke case (1) of Rule 56a, and issue an Opinion conserving A. eucrenophila over A. punctata; and to place the name A. punctata on the list of rejected names. We argue that these actions will give instant stability to a complex and confusing situation by making A. eucrenophila rather than A. punctata the correct name for 'Aeromonas DNA hybridization group 6', an association that is almost universally accepted by the scientific community as reflected in the literature.

Distribution and Potential Effects of 17ß-Estradiol (E2) on Aeromonas Diversity in Wastewater and Fish Samples.

BACKGROUND AND OBJECTIVE: Recently, there has been evidence for the accumulation of steroid hormones in the water environment with negative consequences on fish and humans. However, there is paucity of information on how the steroid hormones influence the microbial community in environmental waters. The objective of this study was to determine the occurrence of 17ß-estradiol (E2) and its potential influence on the diversity of Aeromonas spp. MATERIALS AND METHODS: Wastewater samples were obtained from sewage treatment plants in northern South Africa and fish samples were collected from the Nandoni dam. Aeromonas spp. were isolated using microbiological methods and PCR protocols were used for their identification. A commercial Elisa kit was used for measuring the concentration of 17ß-estradiol (E2) from the wastewater samples as well as the fish samples. RESULTS: 17ß-estradiol (E2) was found in high concentration in sewage samples varying from 0.32-348.6 pg mL-1 while in fish samples, it ranged from 1.1-73.6 pg mL-1. There was a tendency of samples with high E2 concentrations to have higher diversity of Aeromonas spp., implying that steroid hormones may serve as nutrient for Aeromonas spp. Aeromonas hydrophila was the most prevalent species (71%), followed by A. sobria with (68%). CONCLUSION: The presence of Aeromonas spp. in environmental waters and fish that is consumed by the local community poses a serious health concern. The high content of E2 in treated wastewater is of serious concern as well. For the first time, the present study showed a positive impact of E2 on Aeromonas growth.

Characterization of tetracycline effects on microbial community, antibiotic resistance genes and antibiotic resistance of Aeromonas spp. in gut of goldfish Carassius auratus Linnaeus.

The gut of aquatic animals was a significant niche for dissemination of antibiotic resistance genes (ARGs) and direct response of living conditions. In this study, the gut microbiota of goldfish Carassius auratus Linnaeus was sampled at 7 days and 21 days after treatment with tetracycline at 0.285 and 2.85 µg L-1 to investigate the influences on the microbial structure and antibiotic resistance. The proportion of tetracycline resistance bacteria was 1.02% in the control group, while increased to 23.00%, 38.43%, 62.05% in groups of high concentration for 7 days (H7), low concentration for 21 days (L21) and high concentration for 21 days (H21), respectively. Compared to the control group, the diversity of isolated Aeromonas spp. was decreased in the treatment groups and the minimal inhibitory concentration (MIC) of resistant isolates was enhanced from 32 to 256 µg mL-1 with the treatment of tetracycline in time- and dose-dependent manners. Furthermore, the abundance of most genes was increased in treatment groups and efflux genes mainly responded to the stress of tetracycline with an average level of 1.0 × 10-2. After treatment with tetracycline, the predominant species were changed both at phylum and genus levels. The present study explored the impact of tetracycline on gut microbiota of goldfish at environmentally realistic concentrations for the first time and our findings will provide a reference for characterizing the microbiome of fish in the natural environment.