Colistin Resistance Mediated by Mcr-3-Related Phosphoethanolamine Transferase Genes in Aeromonas Species Isolated from Aquatic Environments in Avaga and Pakro Communities in the Eastern Region of Ghana.

Purpose: Colistin is classified by the World Health Organization (WHO) as a critically important and last-resort antibiotic for the treatment of infections caused by carbapenem-resistant bacteria. However, colistin resistance mediated by chromosomal mutations or plasmid-linked mobilized colistin resistance (mcr) genes has emerged. Methods: Thirteen mcr-positive Aeromonas species isolated from water samples collected in Eastern Ghana were analyzed using whole-genome sequencing (WGS). Antimicrobial susceptibility was tested using the broth microdilution method. Resistome analysis was performed in silico using a web-based platform. Results: The minimum inhibitory concentration (MIC) of colistin for all except three isolates was >4 µg/mL. Nine new sequence types were identified and whole-genome analysis revealed that the isolates harbored genes (mcr-3-related genes) that code for Lipid A phosphoethanolamine transferases on their chromosomes. BLAST analysis indicated that the amino acid sequences of the mcr-3-related genes detected varied from those previously reported and shared 79.04-99.86% nucleotide sequence identity with publicly available mcr-3 variants and mcr-3-related phosphoethanolamine transferases. Analysis of the genetic context of mcr-3-related genes revealed that the genetic environment surrounding mcr-3-related genes was diverse among the different species of Aeromonas but conserved among isolates of the same species. Mcr-3-related-gene-IS-mcr-3-related-gene segment was identified in three Aeromonas caviae strains. Conclusion: The presence of mcr-3-related genes close to insertion elements is important for continuous monitoring to better understand how to control the mobilization and dissemination of antibiotic resistance genes.

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.

Prevalence of Extended-Spectrum ß-Lactamases (ESBLs) Producing Aeromonas spp. Isolated from Lamellidens marginalis (Lamark, 1819) of Sewage-Fed Wetland: A Phenotypic and Genotypic Approach.

The global rise of zoonotic bacteria resistant to multiple antimicrobial classes and the growing occurrence of infections caused by Aeromonas spp. resistant to ß-lactam antibiotics pose a severe threat to animal and human health. However, the contribution of natural environments, particularly aquatic ecosystems, as ideal settings for the development and spread of antimicrobial resistance (AMR) is a key concern. Investigating the phenotypic antibiotic resistance and detection of ß-lactamase producing Aeromonas spp. in Lamellidens marginalis, which inhabit all freshwater ecosystems of the Indian subcontinent, is essential for implications in monitoring food safety and drug resistance. In the present investigation, 92 isolates of Aeromonas spp. were recovered from 105 bivalves and screened for their antimicrobial resistance patterns. In vitro antibiotic resistance profiling showed a higher Multiple Antibiotic Resistance (MAR) index of 0.8 with the highest resistance against ampicillin/sulbactam (82%), while 58, 44, 39 and 38% of the isolates were resistant to cephalothin, erythromycin, cefoxitin and imipenem, respectively. PCR results revealed that these isolates carried the blaTEM gene (94%), which was followed by the blaCTX-M gene (51%) and the blaSHV gene (45%). A combination of blaSHV, blaCTX-M, and blaTEM genes was found in 17% of the isolates, indicating the presence of all three resistance genes. This is the first investigation which highlights the importance of multidrug-resistant Aeromonas spp. in L. marginalis. The identification of extended-spectrum-ß-lactamases (ESBLs) genes demand the necessity of continuous surveillance and systematic monitoring, considering its potential health risks for both animals and human beings.

Unveiling novel threats: Urban river isolation of Aeromonas veronii with unusual VEB-28 extended-spectrum ß-lactamase and distinct mcr variants.

Aeromonas spp. are frequently encountered in aquatic environments, with Aeromonas veronii emerging as an opportunistic pathogen causing a range of diseases in both humans and animals. Recent reports have raised public health concerns due to the emergence of multidrug-resistant Aeromonas spp. This is particularly noteworthy as these species have demonstrated the ability to acquire and transmit antimicrobial resistance genes (ARGs). In this study, we report the genomic and phenotypic characteristics of the A. veronii TR112 strain, which harbors a novel variant of the Vietnamese Extended-spectrum ß-lactamase-encoding gene, blaVEB-28, and two mcr variants recovered from an urban river located in the Metropolitan Region of São Paulo, Brazil. A. veronii TR112 strain exhibited high minimum inhibitory concentrations (MICs) for ceftazidime (64 µg/mL), polymyxin (8 µg/mL), and ciprofloxacin (64 µg/mL). Furthermore, the TR112 strain demonstrated adherence to HeLa and Caco-2 cells within 3 h, cytotoxicity to HeLa cells after 24 h of interaction, and high mortality rates to the Galleria mellonella model. Genomic analysis showed that the TR112 strain belongs to ST257 and presented a range of ARGs conferring resistance to ß-lactams (blaVEB-28, blaCphA3, blaOXA-912) and polymyxins (mcr-3 and mcr-3.6). Additionally, we identified a diversity of virulence factor-encoding genes, including those encoding mannose-sensitive hemagglutinin (Msh) pilus, polar flagella, type IV pili, type II secretion system (T2SS), aerolysin (AerA), cytotoxic enterotoxin (Act), hemolysin (HlyA), hemolysin III (HlyIII), thermostable hemolysin (TH), and capsular polysaccharide (CPS). In conclusion, our findings suggest that A. veronii may serve as an environmental reservoir for ARGs and virulence factors, highlighting its importance as a potential pathogen in public health.

Smelly shark, smelly ray: what is infecting you?

AIMS: Although elasmobranchs are consumed worldwide, bacteriological assessments for this group are still sorely lacking. In this context, this study assessed bacteria of sharks and rays from one of the most important landing ports along the Rio de Janeiro coast. METHODS AND RESULTS: Bacteria were isolated from the cloacal swabs of the sampled elasmobranchs. They were cultured, and Vibrio, Aeromonas, and Enterobacterales were isolated and identified. The isolated bacteria were then biochemically identified and antimicrobial susceptibility assays were performed. Antigenic characterizations were performed for Salmonella spp. and Polymerase Chain Reaction (PCR) assays were performed to identify Escherichia coli pathotypes. Several bacteria of interest in the One Health context were detected. The most prevalent Enterobacterales were Morganella morganii and Citrobacter freundii, while Vibrio harveyi and Vibrio fluvialis were the most prevalent among Vibrio spp. and Aeromonas allosacharophila and Aeromonas veronii bv. veronii were the most frequent among Aeromonas spp. Several bacteria also displayed antimicrobial resistance, indicative of Public Health concerns. A total of 10% of Vibrio strains were resistant to trimethoprim-sulfamethoxazole and 40% displayed intermediate resistance to cefoxitin. Salmonella enterica strains displayed intermediate resistance to ciprofloxacin, nalidixic acid and streptomycin. All V. cholerae strains were identified as non-O1/non-O139. The detected E. coli strains did not exhibit pathogenicity genes. This is the first study to perform serology assessments for S. enterica subsp. enterica isolated from elasmobranchs, identifying the zoonotic Typhimurium serovar. Salmonella serology evaluations are, therefore, paramount to identify the importance of elasmobranchs in the epidemiological salmonellosis chain. CONCLUSIONS: The detection of several pathogenic and antibiotic-resistant bacteria may pose significant Public Health risks in Brazil, due to high elasmobranch consumption rates, indicating the urgent need for further bacteriological assessments in this group.

Aeromonas spp. in Freshwater Bodies: Antimicrobial Resistance and Biofilm Assembly.

Aeromonas spp. are environmental bacteria able to infect animals and humans. Here, we aim to evaluate the role of biofilms in Aeromonas persistence in freshwater. Aeromonas were isolated from water and biofilm samples and identified by Vitek-MS and 16S rRNA sequencing. Antibiotic susceptibility profiles were determined according to EUCAST, and a crystal violet assay was used to assess biofilm assembly. MTT and the enumeration of colony-forming units were used to evaluate biofilm and planktonic Aeromonas susceptibility to chlorination, respectively. Identification at the species level was challenging, suggesting the need to improve the used methodologies. Five different Aeromonas species (A. salmonicida, A. hydrophila, A. media, A. popoffii and A. veronii) were identified from water, and one species was identified from biofilms (A. veronii). A. veronnii and A. salmonicida presented resistance to different antibiotics, whith the highest resistance rate observed for A. salmonicida (multiple antibiotic resistance index of 0.25). Of the 21 isolates, 11 were biofilm producers, and 10 of them were strong biofilm producers (SBPs). The SBPs presented increased tolerance to chlorine disinfection when compared with their planktonic counterparts. In order to elucidate the mechanisms underlying biofilm tolerance to chlorine and support the importance of preventing biofilm assembly in water reservoirs, further research is required.

Microbiological and Toxicological Investigations on Bivalve Molluscs Farmed in Sicily.

Bivalves can concentrate biological and chemical pollutants, causing foodborne outbreaks whose occurrence is increasing, due to climatic and anthropic factors that are difficult to reverse, hence the need for improved surveillance. This study aimed to evaluate the hygienic qualities of bivalves sampled along the production and distribution chain in Sicily and collect useful data for consumer safety. Bacteriological and molecular analyses were performed on 254 samples of bivalves for the detection of enteropathogenic Vibrio, Arcobacter spp., Aeromonas spp., Salmonella spp., and beta-glucuronidase-positive Escherichia coli. A total of 96 out of 254 samples, collected in the production areas, were processed for algal biotoxins and heavy metals detection. Bacterial and algal contaminations were also assessed for 21 samples of water from aquaculture implants. Vibrio spp., Arcobacter spp., Aeromonas hydrophila, Salmonella spp., and Escherichia coli were detected in 106/254, 79/254, 12/254, 16/254, and 95/254 molluscs, respectively. A total of 10/96 bivalves tested positive for algal biotoxins, and metals were under the legal limit. V. alginolyticus, A. butzleri, and E. coli were detected in 5, 3, and 3 water samples, respectively. Alexandrium minutum, Dinophysis acuminata, Lingulodinium polyedra, and Pseudonitzschia spp. were detected in water samples collected with the biotoxin-containing molluscs. Traces of yessotoxins were detected in molluscs from water samples containing the corresponding producing algae. Despite the strict regulation by the European Commission over shellfish supply chain monitoring, our analyses highlighted the need for efficiency improvement.

Characterization of Aeromonas Isolates from Ornamental Fish: Species, Virulence Genes, and Antimicrobial Susceptibility.

This study aimed to characterize 300 Aeromonas spp. strains isolated from 123 ornamental fish of 32 different species presenting with septicemia, skin lesions, and/or eye lesions. Within the 300 strains, 53.0% were identified as A. veronii, 41.3% as A. hydrophila, and 5.7% as A. caviae. Among the six virulence genes investigated, the most frequent were act (90.3%) and aer (79.3%). More than 50% of A. hydrophila strains were positive for all the studied genes. A total of 30 virulence profiles were identified, with the five main profiles identified comprising 75% of strains. Only five strains were negative for all genes and were identified as A. caviae and A. veronii. The antimicrobial susceptibility profile was performed for 234 strains, with sulfonamides presenting more than 50% of the resistance rates. Susceptibility was observed mainly for cephalosporins, aminoglycosides, chloramphenicol and piperacillin-tazobactam. Multidrug resistance was detected in 82.5% of the studied strains, including A. caviae with 100% multidrug resistance, and A. hydrophila with 90.9% multidrug resistance. The SE-AFLP analysis resulted in 66 genotypes of A. hydrophila, 118 genotypes of A. veronii, and 14 genotypes of A. caviae, demonstrating the greater heterogeneity of A. veronii and A. caviae. However, no direct correlation was observed between the genotypes and the strains' origins or virulence and resistance profiles.

Detection of chromosome-mediated blaNDM-1-carrying Aeromonas spp. in the intestinal contents of fresh water river fish in Ho Chi Minh City, Vietnam.

The spread of antibiotic-resistant bacteria is a global problem that should be addressed through the perspective of the "one health" concept. The purpose of this study was to determine the contamination rate of antibiotic-resistant Aeromonas spp. in fresh water river fish purchased from a fish market in Vietnam. We then defined the pattern of antibiotic resistance to assess antibiotic-resistant contamination. Antibiotic-resistant Aeromonas spp. were detected in the intestinal contents of 32 of 80 fish. blaNDM-1 was detected in seven strains. Extended-spectrum ß-lactamase and AmpC ß-lactamase-related genes were detected in 28 strains, including blaCTX-M-55, blaCTX-M-15, blaCTX-M-1, and blaDHA,blaFOX, and blaMOX. The blaNDM-1 detected in the seven Aeromonas spp. strains were found chromosomally. This finding suggests that the blaNDM gene is stable in the natural environment and may spread widely into animals and humans via Aeromonas spp. with a transposon. Our results suggest the importance of continuing to monitor carbapenemase genes in Aeromonas spp. to evaluate the possibility that they may spread in other Enterobacterales, and to elucidate the mechanism of spread.

Identification and characterization of motile Aeromonas spp. isolated from farmed Nile tilapia (Oreochromis niloticus) in the Philippines.

AIMS: Motile Aeromonas septicaemia (MAS) caused by motile Aeromonas species is an important disease in farmed freshwater fish due to intensification of culture and improper farm practices. This study characterized and profiled motile Aeromonas species recovered from clinically sick tilapia farmed in the Philippines, with a view to identifying targeted disease prevention and control measures against MAS in farmed tilapia species. METHODS AND RESULTS: Sixteen isolates from diseased farmed Nile tilapia were identified as Aeromonas veronii (n = 14), Aeromonas caviae (n = 1), and Aeromonas dhakensis (n = 1). Five biochemical profiles using API 20E were exhibited by the A. veronii strains giving an unreliable identification. A high level of agreement was observed in identifying the Aeromonas strains using 16S rRNA and rpoD gene sequencing, although the latter has a higher discriminatory value. Three or more virulence genes dominated by cytotoxic enterotoxin act and aerolysin aer were detected. Different genotypes based on virulence gene clustering suggested varied mechanisms used by Aeromonas to colonize and infect or to mutualistically co-exist with the fish. Acquired multiple antibiotic resistance was found in a single A. veronii isolate. All were susceptible to enrofloxacin, oxolinic acid, florfenicol, and chloramphenicol. Tetracycline and sulfonamide resistances and class 1 integron were detected in three A. veronii isolates. CONCLUSION: Several strains of motile aeromonads, especially A. veronii, which have varied genotypes based on virulence, biochemical profile, and antibiotic resistance, are involved in MAS in natural disease outbreaks in farmed Nile tilapia in the Philippines.

Municipal wastewater monitoring revealed the predominance of bla GES genes with diverse variants among carbapenemase-producing organisms: high occurrence and persistence of Aeromonas caviae harboring the new bla GES variant bla GES-48.

IMPORTANCE: The emergence and spread of carbapenemase-producing organisms (CPOs) represent a global health threat because they are associated with limited treatment options and poor clinical outcomes. Wastewater is considered a hotspot for the evolution and dissemination of antimicrobial resistance. Thus, analyses of municipal wastewater are critical for understanding the circulation of these CPOs and carbapenemase genes in local communities, which remains scarcely known in Japan. This study resulted in several key observations: (i) the vast majority of bla GES genes, including six new bla GES variants, and less frequent bla IMP genes were carbapenemase genes encountered exclusively in wastewater influent; (ii) the most dominant CPO species were Aeromonas spp., in which a remarkable diversity of new sequence types was observed; and (iii) CPOs were detected from combined sewer wastewater, but not from separate sewer wastewater, suggesting that the load of CPOs from unrecognized environmental sources could greatly contribute to their detection in influent wastewater.

Phenotypic and genotypic characteristics of beta-lactamase dominant with CARBA, AmpC, and ESBL-producing bacteria in municipal wastewater influent in Helsinki, Finland.

OBJECTIVES: Analysing samples of municipal wastewater influent (before treatment) can help to map the status of antibiotic-resistant bacteria (ARB) at the population level in sewershed communities and may also help in predicting the public health risks of ARB in surface water because of the outfall of wastewater. In this study, we investigated the bacterial isolates carrying beta-lactamase genes in wastewater and compared their genotypic and phenotypic characteristics. METHODS: A total of 399 bacterial isolates grown on CHROMagarESBL (n = 207) and CHROMagarKPC (n = 192) from composite wastewater influent samples (n = 7) from the Viikinmäki wastewater treatment plant (Helsinki) were subcultured, nucleic acid was extracted, and the prevalence of different beta-lactamase genes was screened with multiplex polymerase chain reaction (PCR). All PCR-positive isolates were identified with MALDI-TOF. RESULTS: A total of 32.6% of isolates (130 of 399) were PCR positive for at least one resistance gene, and 13% of these positive isolates out of 130 had at least three resistance genes. Among the 22 detected genes, blaGES group was the most prevalent, at 25.8% (n = 198; many isolates carried multiple genes), followed by blaMOX (13.1%) and blaTEM (10.1%) as most frequently detected. Furthermore, out of 18 different bacterial species/genera detected as carrying beta-lactamase genes, A. hydrophila/caviae (28.5%), Enterobacter spp. (16.9%), and E. coli (14.6%) were the most prevalent. Enterobacter spp., Aeromonas spp., and K. cryocescens potentially carried AmpC genes, and E. coli carried ESBL genes. CONCLUSION: We recorded a huge variety of beta-lactamases (blaAmpC, blaESBL, and blaCARBA) genes in many potential pathogens that probably originated from both enteric and environmental sources.

Epidemiology of Aeromonas spp. isolated from stool in a tertiary hospital in Cantabria, Northern Spain, in the last five years / Epidemiología de Aeromonas spp. aisladas de heces en un hospital terciario en Cantabria, norte de España, en los últimos 5 años

Introduction: The role of Aeromonas species in gastrointestinal disease is controversial. The aim of this study was to know the epidemiological distribution of Aeromonas spp. isolated from stool in our health area, determine the existence of diarrhea as a significant symptom, identification of existing species in our environment and association as co-pathogen. Methods: It was a retrospective descriptive study of isolates of Aeromonas spp. in feces (2016–2020). The protocol for these isolates included coproculture, identification by MALDI-TOF (Vitek-MS®, BioMerieux) and confirmation by multiplex PCR. Results: A total of 366 Aeromonas spp. isolates were analyzed being Aeromonas caviae the most prevalent species (289, 78.7%). A total of 58 (15.8%) co-infections were identified, being more frequent in pediatric age (49;84.5%) (p=0.01) and mostly associated with Campylobacter spp. Discussion: Aeromonas spp. prove to be a gastrointestinal pathogen more frequently associated with co-infections in pediatric age, evidencing its appearance especially with Campylobacter spp.(AU)

Introducción: El papel de las especies de Aeromonas en la enfermedad gastrointestinal es muy controvertido. El objetivo de este estudio fue conocer la distribución epidemiológica de Aeromonas spp. aislada de heces en nuestra área de salud, determinar la existencia de diarrea como síntoma significativo, la identificación de especies existentes en nuestro entorno y la asociación como copatógeno. Métodos: Se trata de un estudio descriptivo retrospectivo de aislados de Aeromonas spp. en heces (2016-2020). El protocolo para estos aislamientos incluía coprocultivo, la identificación por MALDI-TOF (VITEK®MS, bioMérieux) y la confirmación por PCR multiplex. Resultados: Se analizaron un total de 366 aislados de Aeromonas spp., siendo Aeromonas caviae la especie más prevalente (289; 78,7%). Se identificaron un total de 58 (15,8%) coinfecciones, siendo significativamente más frecuentes en edades pediátricas (49; 84,5%) (p=0,01) y asociadas principalmente con Campylobacter spp. Discusión: Aeromonas spp. resulta ser un patógeno gastrointestinal que se asocia con mayor frecuencia a coinfecciones en la edad pediátrica, evidenciando su aparición especialmente con Campylobacter spp.(AU)

Antibiotic Resistance, Virulence Gene Detection, and Biofilm Formation in Aeromonas spp. Isolated from Fish and Humans in Egypt.

The genus Aeromonas is widely distributed in aquatic environments and is recognized as a potential human pathogen. Some Aeromonas species are able to cause a wide spectrum of diseases, mainly gastroenteritis, skin and soft-tissue infections, bacteremia, and sepsis. The aim of the current study was to determine the prevalence of Aeromonas spp. in raw fish markets and humans in Zagazig, Egypt; identify the factors that contribute to virulence; determine the isolates' profile of antibiotic resistance; and to elucidate the ability of Aeromonas spp. to form biofilms. The examined samples included fish tissues and organs from tilapia (Oreochromis niloticus, n = 160) and mugil (Mugil cephalus, n = 105), and human skin swabs (n = 51) and fecal samples (n = 27). Based on biochemical and PCR assays, 11 isolates (3.2%) were confirmed as Aeromonas spp. and four isolates (1.2%) were confirmed as A. hydrophila. The virulence genes including haemolysin (hyl A) and aerolysin (aer) were detected using PCR in A. hydrophila in percentages of 25% and 50%, respectively. The antimicrobial resistance of Aeromonas spp. was assessed against 14 antibiotics comprising six classes. The resistance to cefixime (81.8%) and tobramycin (45.4%) was observed. The multiple antibiotic resistance (MAR) index ranged between 0.142-0.642 with 64.2% of the isolates having MAR values equal to 0.642. Biofilm formation capacity was assessed using a microtiter plate assay, and two isolates (18.1%) were classified as biofilm producers. This study establishes a baseline for monitoring and controlling the multidrug-resistant Aeromonas spp. and especially A. hydrophila in marine foods consumed in our country to protect humans and animals.

Occurrence of potential virulence determinants in Aeromonas spp. isolated from different aquatic environments.

Aeromonas species are bacterial pathogens that cause significant economic losses in the aquaculture industry worldwide. They are widely distributed in aquatic environments and cause several diseases in both humans and aquatic animals. The presence of various virulent Aeromonas spp. in aquatic environments predisposes infections in aquatic animals and humans. Concern about the transfer of pathogens from fish to humans also increased with the substantial increase in seafood consumption. Aeromonas spp. are also primary human pathogens that cause local and systemic infections, both in immunologically compromised and immunologically competent hosts. The most common Aeromonas spp. causing infections in aquatic animals and humans are A. hydrophila, A. salmonicida, A. caviae, and A. veronii biotype sobria. The ability of Aeromonas spp. to produce a variety of virulence factors enhances their pathogenic ability. Literature has supplied evidence for the presence of various virulence factors, including proteases, enterotoxins, hemolysin, and toxin genes of Aeromonas spp. in aquatic environments. The high prevalence of Aeromonas spp. in the aquatic environment is also a threat to public health. Because the Aeromonas spp. infections in humans are generally the result of ingestion or exposure to contaminated food and water. This review summarizes the recently published information on various virulence factors and virulence genes of Aeromonas spp. isolated from various aquatic environments, including seawater, freshwater, wastewater, and drinking water. It is also intended to highlight the risks associated with Aeromonas species' virulence properties for both aquaculture and public health.

Virulence genes of Aeromonas spp. isolates from stool in Spain.

INTRODUCTION: The role of Aeromonas species in gastrointestinal disease is controversial. The aim was to analyze not only the virulence genes between different species of Aeromonas isolated from feces, but the distribution of these virulence genes between enterotoxigenic strains and co-pathogen strains. METHODS: Retrospective study of isolates of Aeromonas spp. in feces (2016-2021). The protocol included coproculture, identification by MALDI-TOF and confirmation by multiplex PCR. SPSS Statistics program was used. RESULTS: A total of 288 strains were studied for the virulence genes between different species of Aeromonas. To compare virulence genes between Aeromonas as co-pathogen and those isolated alone, 218 strains of the global set were used; 52 as co-pathogens compared with 166 Aeromonas without associated pathogen as controls. CONCLUSIONS: We found no significant differences in the distribution of virulence genes versus co-existence of co-pathogens or not. A. hydrophila is the potentially most virulent species of our set.

Epidemiology of Aeromonas spp. isolated from stool in a tertiary hospital in Cantabria, Northern Spain, in the last five years.

INTRODUCTION: The role of Aeromonas species in gastrointestinal disease is controversial. The aim of this study was to know the epidemiological distribution of Aeromonas spp. isolated from stool in our health area, determine the existence of diarrhea as a significant symptom, identification of existing species in our environment and association as co-pathogen. METHODS: It was a retrospective descriptive study of isolates of Aeromonas spp. in feces (2016-2020). The protocol for these isolates included coproculture, identification by MALDI-TOF (Vitek-MS®, BioMerieux) and confirmation by multiplex PCR. RESULTS: A total of 366 Aeromonas spp. isolates were analyzed being Aeromonas caviae the most prevalent species (289, 78.7%). A total of 58 (15.8%) co-infections were identified, being more frequent in pediatric age (49;84.5%) (p=0.01) and mostly associated with Campylobacter spp. DISCUSSION: Aeromonas spp. prove to be a gastrointestinal pathogen more frequently associated with co-infections in pediatric age, evidencing its appearance especially with Campylobacter spp.

Uses of Antibiotics in Ornamental Fish in Hong Kong and the Antibiotic Resistance in the Associated Zoonotic Pathogens.

The use of antibiotics in ornamental fish is not regulated, as they are not intended for human consumption. Although antibiotic resistant bacteria have been detected in ornamental fish worldwide, there have been no studies to look at the situation in Hong Kong. Therefore, the present study was conducted to investigate the use of antibiotics in ornamental fish. Ornamental fish were purchased from five local pet fish shops and the antibiotics in carriage water were quantified using liquid chromatography tandem mass spectrometry. Moreover, Aeromonas and Pseudomonas spp. present in carriage water were isolated and their minimum inhibitory concentrations against selected antibiotics were determined. Results indicated that among the twenty antibiotics screened, doxycycline (0.0155-0.0836 µg L-1), oxytetracycline (0.0102-29.0 µg L-1), tetracycline (0.0350-0.244 µg L-1), enrofloxacin (0.00107-0.247 µg L-1), and oxalinic acid (n.d.-0.514 µg L-1) were detected in all sampled shops. Additionally, MIC results revealed that some of the Aeromonas and Pseudomonas spp. isolates were highly resistant to all antibiotics selected. Our findings confirmed that multiple antibiotics are being used in ornamental fish and the associated bacteria are resistant to selected antibiotics, suggesting that this could be a significant transmission route of antibiotic resistant bacteria to household indoor environments.

Occurrence, antimicrobial resistance, virulence, and biofilm formation capacity of Vibrio spp. and Aeromonas spp. isolated from raw seafood marketed in Bangkok, Thailand.

Background and Aim: Bacteria of the genera Vibrio and Aeromonas cause seafood-borne zoonoses, which may have a significant impact on food safety, economy, and public health worldwide. The presence of drug-resistant and biofilm-forming phenotypes in the food chain increases the risk for consumers. This study aimed to investigate the characteristics, virulence, biofilm production, and dissemination of antimicrobial-resistant pathogens isolated from seafood markets in Bangkok, Thailand. Materials and Methods: A total of 120 retail seafood samples were collected from 10 local markets in Bangkok and peripheral areas. All samples were cultured and the Vibrio and Aeromonas genera were isolated using selective agar and biochemical tests based on standard protocols (ISO 21872-1: 2017). The antibiotic susceptibility test was conducted using the disk diffusion method. The presence of hemolysis and protease production was also investigated. Polymerase chain reaction (PCR) was used to determine the presence of the hlyA gene. Furthermore, biofilm formation was characterized by microtiter plate assay and scanning electron microscopy. Results: The bacterial identification test revealed that 35/57 (61.4%) belonged to the Vibrio genus and 22/57 (38.6%) to the Aeromonas genus. The Kirby-Bauer test demonstrated that 61.4% of the isolates were resistant to at least one antibiotic and 45.61% had a high multiple antibiotic resistance index (≥0.2). PCR analysis indicated that 75.44% of the bacteria harbored the hlyA gene. Among them, 63.16% exhibited the hemolysis phenotype and 8.77% showed protease activity. The biofilm formation assay demonstrated that approximately 56.14% of all the isolates had the potential to produce biofilms. The moderate biofilm production was the predominant phenotype. Conclusion: The results of this study provide evidence of the multiple drug resistance phenotype and biofilm formation capacity of Vibrio and Aeromonas species contaminating raw seafood. Effective control measures and active surveillance of foodborne zoonoses are crucial for food safety and to decrease the occurrence of diseases associated with seafood consumption.