Aeromonas Hydrophila Isolates Research Articles

First Report and Characterization of a Plasmid-Encoded blaSFO-1 in a Multi-Drug-Resistant Aeromonas hydrophila Clinical Isolate.

Antibiotic resistance remains one of the most pressing public health issues facing the world today. At the forefront of this battle lies the ever-increasing identification of extended-spectrum beta-lactamases and carbapenemases within human pathogens, conferring resistance towards broad-spectrum and last-resort antimicrobials. This study was prompted due to the identification of a pathogenic Aeromonas hydrophila isolate (strain MAH-4) collected from abdominal fluid, which presented a robust resistance pattern against second-, third-, and fourth-generation cephalosporins, ertapenem, ciprofloxacin, gentamicin, levofloxacin and moxifloxacin, and beta lactam/beta-lactamase inhibitor combinations. Whole genome sequencing was performed and identified a 328 kb plasmid (pMAH4) encoding 10 antibiotic resistance genes, including blaSFO-1, blaTEM-1, and blaOXA-1 of A. hydrophia MAH-4. This is the first report of beta-lactamase SFO-1 within a clinical strain of Aeromonas. Due to the remarkable sequence identity of pMAH4 to plasmids associated with Enterobacterales genera like Klebsiella and the extensive capabilities of Aeromonas for horizontal gene transfer, our identification of a clinical isolate encoding SFO-1 on a plasmid suggests antibiotic resistance gene mobility between Enterobacterales and non-Enterobacterales species.

Molecular characterization of Aeromonas hydrophila isolates from diseased fishes in district Kasur, Punjab, Pakistan

Abstract Pakistan is an agricultural country and fisheries play a very important role in the economic development of the country. Different diseases are prevalent in Pakistani fish but information related to the causative agents is not well-known. Keeping in view the significance of bacterial pathogens as the causative agents of multiple fish diseases, the present study was conducted for identification, characterization and analysis of virulence genes of Aeromonas spp. isolated from diseased fishes. A total of fifty fish samples having multiple clinical indications were collected from different fish farms of district Kasur, Punjab Pakistan. For isolation of Aeromonas spp. samples were enriched and inoculated on Aeromonas isolation medium. Isolates were identified and characterized by different biochemical tests, Analytical Profile Index (API) 20E kit and Polymerase Chain Reaction (PCR) assays. All isolates were screened for three putative virulence genes including aerolysin (aer), haemolysin (hyl) and heat labile cytotonic enterotoxin (alt). Seven isolates of Aeromonas (A.) hydrophila were retrieved and identified based on API 20E. These isolates were further confirmed as A. hydrophila on the basis of PCR assays. Three isolates were detected positive for the presence of virulence genes (alt and hyl). Whereas aerolysin (aer) gene was not present in any of A. hydrophila isolates. The present study confirmed A. hydrophila as the causative agent of epizootic ulcerative syndrome and motile Aeromonas septicemia in fish farms of district Kasur, Punjab Pakistan. Moreover, detection of two virulence genes (alt and hyl) in A. hydrophila isolates is a threat for fish consumers of study area.

Clinical and pathological investigations in ulcer disease of Cyrinus carpio caused by Aeromonas hydrophila

The Indian carps are the most important food fishes and they have a considerable economic impact on the country's economy. In several parts of the country, they are widely cultivated. The Indian Major Carps include the Catla catla, Labeo calbasu, Cirrhinus mrigala, and Labeo rohita, to name a few species. The fish Cyprinus carpio, both in the wild and in captivity, is vulnerable to Aeromonas hydrophila infections. Investigations carried out on Cyrinus carpio to better understand the implications of a pathogenic Aeromonas hydrophila isolates impacts on fish health revealed that the pathogen affects severely. The fish species that were injected with the bacterial suspension developed lesions of a similar nature, and dead fishes with ulcers on their bodies were found in all of the tanks. Even after 15 days of incubation and observation, no difference was observed in the fishes in the control group of fishes. Despite this, not all of the fish who got ulcers perished. Healing has been shown in some fish that had small ulcers, according to the research. Cyprinus carpio injected with a mixed bacterial suspension of AH-04 and AH-07 recorded a death rate was 65.0 percent.

Phenotypic characterization, genetic analysis and antibiotic sensitivity of Aeromonas hydrophila isolates causing dropsy in cultured Labeo rohita from Punjab, Pakistan

Motile Aeromonas septicemia (MAS) is a common freshwater fish disease and major threat to the aquaculture in Pakistan. The present study was carried out on suspected fish samples to isolate and characterize local strains of Aeromonas hydrophila, a key pathogen responsible for the said disease in aquacultured fishes. A total of ninety suspected fish specimens were collected from fish farms in Kasur, Okara and Gujranwala districts of Punjab, Pakistan from June 2018 to April 2019. The specimens were processed and A. hydrophila strains were isolated. The primary identification of sixty seven isolates were verified by colony morphology, microscopy and phenotypic characterization with ten biochemical reactions. The A. hydrophila strains of test samples were molecularly characterized by polymerase chain reaction (PCR) using 16S rRNA at desired size of 356bp. The PCR amplified product was subjected to DNA sequencing and phylogenetic analysis showed homology with related strains of Aeromonas spp. By antibiotic sensitivity test, the isolates were checked for nine antibiotics in which the pathogen was sensitive to four and resistant to five drugs. Results of genetic analysis confirmed strains as A. hydrophila which are useful to take preventive measures against the said disease.

Detection and distribution of virulence genes in Aeromonas hydrophila isolates causing infection in cultured carps.

Aeromonas hydrophila is a bacterium associated with many diseases and disorders such as fin rot, skin ulcers and lethal hemorrhagic septicemia in fish. It bears several virulence factors including type III secretion system (T3SS), aerolysin, cytolytic enterotoxin and enzymes (e.g., hemolysins, lipase) that seem to play an important role in its pathogenesis. Detection of virulence markers by polymerase chain reaction (PCR) is a key procedure in defining the patho-genic ability of pathogenic bacteria and preparing a vaccine for its treatment. In this sense, this study was aimed to determine the frequency of virulence genes in isolates obtained from infected cultured carps in Khuzestan province. Out of 200 moribund carps with septicemic symptoms, 125 isolates were belonged to the motile aeromonads and 59 isolates were identified as A. hydrophila by biochemical methods. Finally, using PCR analysis, 31 isolates were identified as A. hydrophila. Five virulence genes were detected in these isolates including hemolysin, aerolysin, cytolytic enterotoxin and T3SS (aopB and ascV) by specific primers. Results showed that 23 (74.19%), 18 (58.06%), 16 (51.61%), 13 (41.63%) and 10 (32.25%) isolates possessed cytolytic enterotoxin, hemolysin, aerolysin, and T3SS genes, respectively. The results of the present study showed that among 31 isolates, only five isolates had all of dominant virulence genes. Thirteen other isolates had genotypes including hlyA+, aerA+, and act+. The remaining isolates had at least one virulence gene. This study showed that determination of the virulence genes by PCR can be a reliable method to identify a potential pathogenic Aeromonad strain.

Purification and structural characterization of lectin with antibacterial and anticancer properties from grubs of hide beetle, Dermestes frischii

Lectins or haemagglutinins are diverse classes of non-immune proteins; they bind to carbohydrates and are abundant in nature. In the present study, a coleopteran lectin from grubs of hide beetle, Dermestes frischii called DFL, was purified by glutaraldehyde (fixative-agent) fixed hen erythrocytes and characterized further for its functional properties. The purified DFL was stable between pH range 5 to 9 and heat-stable up to 50 °C. It was insensitive to EDTA and did not require any divalent cations. DFL native molecular mass was approximately 69 kDa with three different polypeptide subunits of 33 (pI ~4.4), 22 (pI ~6) and 14 (pI ~4.4) kDa. Haemagglutinating activity of DFL was highly inhibited by N-acetyl-D-glucosamine. DFL partial peptide sequences obtained from peptide mass fingerprinting experiments matched with amino acid sequences of lectins from different organisms confirmed its nature. Biological properties of purified DFL namely antibacterial and bacterial agglutination experiments revealed that DFL have both the effects against laboratory cultures of Aeromonas hydrophila, Enterococcus faecalis, Escherichia coli and habitat bacterial isolates of Staphylococcus cohnii and Bacillus cereus. In addition, the DFL exhibited substantial anticancer properties against HeLa cells. These results concluded that purified DFL could serve as a potent therapeutic agent for various biomedical applications.

Gut microbiota-mediated ferroptosis contributes to mercury exposure-induced brain injury in common carp.

Mercury is a heavy metal, which causes irreversible toxicity to fish and is found in aquatic environments around the world. The purpose of this study was to investigate the relative mechanism of mercury exposure on brain injury in common carp. The results showed that mercury exposure could induce brain injury and memory loss in common carp. Meanwhile, mercury exposure could induce neuronal ferroptosis. The ferroptosis inhibitor ferrostatin-1 attenuated mercury-induced brain injury. However, in an in vitro study, mercury did not induce ferroptosis, and ferrostatin-1 did not attenuate mercury-induced common carp brain cell death. Therefore, we speculated that mercury exposure-induced ferroptosis might occur through other pathways. Studies have shown that the gut microbiota contributes to the pathological process of heavy metal-induced injury. Therefore, we detected the effects of mercury exposure on the gut microbiota composition. The results showed that the composition and diversity of the gut microbiota were affected by mercury chloride. Surprisingly, we found that the abundance of Aeromonas, one of the most important pathogenic bacteria of fish, increased significantly. Subsequently, we isolated Aeromonas hydrophila from mercury-exposed carp and these bacteria could lead to brain injury and ferroptosis in common carp. These results suggested that mercury exposure-induced brain injury partly by increasing intestinal A. hydrophila, which led to ferroptosis in common carp.

Molecular and Functional Characterization of a Novel Plasmid-Borne bla NDM-Like Gene, bla AFM-1, in a Clinical Strain of Aeromonas hydrophila.

PurposeAn increasing frequency of antibiotic resistance has been observed in both clinical and environmental Aeromonas hydrophila isolates in recent years. However, there are still very few in-depth studies regarding the role of plasmids in the antibiotic resistance of A. hydrophila. Hence, we investigated the molecular and functional characterization of a multidrug-resistant plasmid encoding an NDM-like metallo-β-lactamase, AFM-1, in the clinical A. hydrophila isolate SS332.MethodsThe minimum inhibitory concentrations (MICs) of 24 antibiotics against A. hydrophila SS332 were measured by the agar dilution method. The genome of A. hydrophila SS332 was sequenced with PacBio and Illumina platforms. Six plasmid-borne antimicrobial resistance genes were chosen for cloning, including blaAFM-1, blaOXA-1, msr(E), mph(E), aac(6ʹ)-Ib10, and aph(3ʹ)-Ia. Phylogenetic analysis, amino acid sequence alignment, and comparative genomic analysis were performed to elucidate the active site requirements and genetic context of the blaAFM-1 gene.ResultsA. hydrophila SS332 showed high levels of resistance to 15 antibiotics, especially those with MIC levels at or above 1024 μg/mL, including ampicillin, cefazolin, ceftriaxone, aztreonam, spectinomycin, and roxithromycin. Six plasmid-borne resistance genes from A. hydrophila were verified to be functional in E. coli DH5α. AFM-1 shared 86% amino acid identity with NDM-1 and showed resistance to ampicillin, cefazolin, cefoxitin, and ceftazidime. In addition, the blaAFM-1 gene was associated with three different novel ISCR19-like elements, designated ISCR19-1, ISCR19-2 and ∆ISCR19-3, which may be involved in the acquisition and mobilization of the blaAFM-1 gene.ConclusionOur investigation showed that plasmid-borne resistance genes can contribute to antibiotic resistance in A. hydrophila SS332. A novel blaNDM-like gene, blaAFM-1, was verified to be functional and associated with novel ISCR19-like elements. This fact indicated the risk of spread of blaAFM-1 genes and ISCR19-like elements.

Prevalence and virulence characteristics of Aeromonas species isolated from fish farms in Egypt

Background/Objective: Fish is one of the most valuable food for human consumption. However, fish may also act as a source of foodborne pathogens including Aeromonas species which caused a serious threat to a public health concern. This study aimed to investigate the prevalence and virulence characteristics of Aeromonas species isolated from fish farms in Egypt. Methods: A total of 100 random samples of freshwater fish represented by Nile tilapia and Mugil cephalus (50 of each) were collected from different fish farms in Kafr El-Sheikh governorate and examined bacteriologically and biochemically for the presence of Aeromonas species. Multiplex PCR was done to detect some virulence-associated genes in Aeromonas hydrophila isolates. Results: The obtained results revealed that the incidence of Aeromonas species in Nile tilapia and Mugil cephalus were 32 (64%) and 25 (50%), respectively. The most prevalent Aeromonas species isolated from Nile tilapia were Aeromonas caviae 13 (40.6%), Aeromonas hydrophila 8 (25%), Aeromonas sobria 7 (21.9%), Aeromonas veronii 3 (9.4%), and Aeromonas fluvialis 1 (3.1%), while from fresh Mugil cephalus were Aeromonas sobria 11 (44%), Aeromonas caviae 7 (28%), Aeromonas hydrophila 5 (20%), and Aeromonas veronii 2 (8%). The isolated Aeromonas hydrophila concealed some virulence genes responsible for their pathogenicities such as aerolysin gene (aerA) and cytotonic enterotoxin gene (altA). Conclusion: The hygienic measures should be applied to prevent fish contamination with Aeromonas species.

Molecular characterization and immunohistochemical localization of tilapia piscidin 3 in response to Aeromonas hydrophila infection in Nile tilapia.

The antimicrobial activity of tilapia piscidin 3 (TP3) was determined in vitro against a locally isolated Aeromonas hydrophila. A 388 bp fragment was amplified from the TP3 cDNA and sequenced. The coding sequence (CDS) of TP3 was estimated to be 231 bp codes for 76 amino acids long and stop codon. In silico analysis was performed to detect both the signal peptide and the prodomain cleavage sites to follow the amino acids number 22 and 70, respectively. Based on this, a peptide 23 amino acids long with a remarkably high computed antimicrobial probability was synthesized and used in the subsequent experiments. The antimicrobial activity of TP3 was determined with minimum inhibitory concentration (MIC) and minim um bactericidal concentration (MBC) methods. TP3 exhibited relatively weak antimicrobial activities against the tested bacteria. A challenge experiment was then performed in Nile tilapia with low and high doses of A. hydrophila, followed by timely recognition; after 3, 6, 24 h, and 7 days of the specific TP3 gene expression, immunohistochemical localization was also performed. Histopathological examination revealed provoked inflammatory responses and congestion in the same organs of TP3 expression. Immunohistochemical localization showed that A. hydrophila induced tilapia fish to express TP3 after 24 h within the gills, intestine, hepatopancreas, spleen, and posterior kidney. In quantitative real time (RT)-polymerase chain reaction analysis, the high dose showed higher mRNA expression levels than the low dose, and its expression levels increased in the A. hydrophila-infected fish. It was therefore concluded that TP3 plays an essential role in fish immunity.

Diversity of virulence-associated genes in pathogenic Aeromonas hydrophila isolates and their in vivo modulation at varied water temperatures

Most environmental parameters have no consistent effect on the expression of bacterial genes responsible for their virulence. However, as fish are poikilothermic, the possibility of temperature variation having a pronounced effect on the expression of virulence-associated gene(s) of bacteria infecting the host needs to be investigated. In this study, the diversity of virulence genes in seven Aeromonas hydrophila isolates collected from diseased fish from different parts of India was characterized, and the effect of temperature variation on the extent of expression of their virulence was investigated. All bacterial isolates were screened for a total of nine bacterial virulent genes {aerolysin, hemolysin, cytoen, outer membrane protein TS (Omp TS), elastase, flagellin, lipase, β hemolysin and type 3 secretion system}, and the diversity in their presence or absence were marked at a particular in vitro condition. Three bacterial isolates (nos. 1, 7 and 2) were selected for further study, based on their ability to cause varied mortalities (20–100%) in Labeo rohita juveniles in intraperitoneal challenge study. Further, three isolates were injected intraperitoneally into L. rohita fingerlings at three different temperatures (i.e., 20, 28 and 37 °C) and at 6 h post-challenge, the kidney samples were collected to measure the levels of all nine bacterial virulence genes using semi-quantitative PCR. The maximum level of amplicons of virulence genes in all three A. hydrophila isolates was noticed at 28 °C as compared to 37 °C and 20 °C. It was also observed that haemolysin played a more prominent role in the expression of virulence, when compared to cytoen gene. Hence, it was concluded that water temperature does play a crucial role in governing virulence gene expression, and a temperature of 28 °C would be considered as suitable for looking into the pathogenicity of A. hydrophila for conducting any challenge study with this organism in tropical environment.

The Isolation of Antimicrobial Resistant Aeromonas spp. from Aquaculture Samples and their Susceptibility to Medicinal Plant Extracts

Aeromonas spp, ubiquitous in both terrestrial and aquatic environments are becoming renowned as enteric pathogens of serious public health concern as they have a number of virulence and resistant determinants that are linked to both human and aquatic diseases due to consistent and incorrect use of antibiotics in aquaculture. The effect of crude aqueous and ethanol extracts of some medicinal plants on antimicrobial resistant Aeromonas spp. isolated from aquaculture water and fish samples was studied. Two hundred and forty (240) Aeromonas isolates, made up of 168 Aeromonas hydrophila and 72 Aeromonas salmonicida, were recovered from aquaculture water and fish gill samples collected from different commercial fish ponds using selective media. The isolates were assessed for their antibiotic susceptibility against ten (10) conventional antibiotics using the Kirby-Bauer technique. Extracts from three medicinal plants, Vernonia amygdalina, Ocimum gratissimum and Garcinia kola were also analyzed for their antimicrobial effects on the isolates that were resistant to the conventional antibiotics. Aeromonas hydrophila isolates expressed the highest resistant rates of 100%, 78.6% and 70.8% to aztreonam, cefotaxime and neomysin respectively, and the A. salmonicida isolates also had a similar trend of high resistant rates of 100%, 87.5% and 77.8% to aztreonam, neomysin and cefotaxime respectively. Antimicrobial resistant analyses with the plant extracts showed 100% inhibition of the isolates at 100 mg/ml for both aqueous and ethanol extracts. Phytochemical screening identified the presence of certain phytochemicals like alkaloids, saponins, tannins, flavonoids, steroids and glycosides which could have accounted for the antimicrobial effects of the plant extracts under study. It can be inferred then, that extracts from Vernonia amygdalina, Ocimum gratissimum and Garcinia kola can inhibit resistant aquaculture Aeromonas isolates and so can present an alternative source of antimicrobials in the effort to combat the increasing incidence of antimicrobial resistance in aquaculture.

Virulence-Associated Genes and Antimicrobial Resistance of Aeromonas hydrophila Isolates from Animal, Food, and Human Sources in Brazil.

Aeromonads are natural inhabitants of aquatic environments and may be associated with various human or animal diseases. Its pathogenicity is complex and multifactorial and is associated with many virulence factors. In this study, 110 selected Aeromonas hydrophila isolates isolated from food, animals, and human clinical material from 2010 to 2015 were analyzed. Antimicrobial susceptibility testing was performed by the disk diffusion method, and polymerase chain reaction was conducted to investigate the virulence genes hemolysin (hlyA), cytotoxic enterotoxin (act), heat-labile cytotonic enterotoxin (alt), aerolysin (aerA), and DNase-nuclease (exu). At least 92.7% of the isolates had one of the investigated virulence genes. Twenty different virulence profiles among the isolates were recognized, and the five investigated virulence genes were observed in four isolates. Human source isolates showed greater diversity than food and animal sources. Antimicrobial resistance was observed in 46.4% of the isolates, and multidrug resistance was detected in 3.6% of the isolates. Among the 120 isolates, 45% were resistant to cefoxitin; 23.5% to nalidixic acid; 16.6% to tetracycline; 13.7% to cefotaxime and imipenem; 11.8% to ceftazidime; 5.9% to amikacin, gentamicin, and sulfamethoxazole-trimethoprim; and 3.9% to ciprofloxacin and nitrofurantoin. Overall, the findings of our study indicated the presence of virulence genes and that antimicrobial resistance in A. hydrophila isolates in this study is compatible with potentially pathogenic bacteria. This information will allow us to recognize the potential risk through circulating isolates in animal health and public health and the spread through the food chain offering subsidies for appropriate sanitary actions.

Desulfonation of the textile azo dye Acid Fast Yellow MR by newly isolated Aeromonas hydrophila SK16

Acid Fast Yellow MR (AFYMR), a sulphonated reactive textile azo dye, was effectively decolorized by a newly identified bacterial strain Aeromonas hydrophila SK16. Nineteen isolates were obtained from the textile effluent polluted soil in Perundurai region. Maximum decolorization of AFYMR was observed in Aeromonas hydrophila SK16. Enzyme activities present in A. hydrophila SK16 revealed the involvement of oxidoreductase and efficiently decolorized 91.25% of AFYMR (100 mg L-l) in static condition for 9 h. Fourier Transform Infrared Spectroscopy (FTIR) analysis showed that A. hydrophila SK16 cells preferred N–H and Sulphur dioxide bonds in the breakdown of AFYMR. The final metabolite supports the degradation of AFYMR through desulfonation as revealed by Gas Chromatography-Mass Spectroscopy (GCMS) results. Phytotoxicity studies conducted in Oryza sativa and Zea mays illustrated significant decrease in toxicity of metabolite produced during decolorization of Acid Fast Yellow MR by Aeromonas hydrophila SK16.

Diversity in gene arrangement in a DNA region lacking aerA in clinical and environmental Aeromonas hydrophila isolates.

Aquatic pathogen Aeromonas hydrophila produces an array of virulence factors, many of which are excreted proteins that causes infectious disease in fish, reptiles, and humans. Aerolysin, a haemolytic toxin, is the most well-known of the A. hydrophila virulence factors and is encoded by aerA. Although used as a virulence gene marker in several studies, recent whole-genome sequencing data suggest there may be some variation in aerolysin genes, as well as in the genetic environment of these genes, among A. hydrophila strains. Here, we used PCR-based assays to examine gene arrangement in the traditional aerA region of 42 aerA-minus clinical and environmental A. hydrophila isolates. PCR primers were designed based on known genes from within the target regions of reference strains carrying non-aerA aerolysin genes. Analyses revealed four different gene arrangement patterns among the isolates, indicating considerable genetic diversity in the target region. While 19 of the 21 environmental isolates showed the same gene pattern, all four patterns were represented among the clinical isolates, implying that the gene pattern is highly conserved in the target region among environmental isolates. Further analysis of the gene regions showed that the predominant pattern among environmental isolates, which did not contain an aerolysin gene, appeared to be the progenitor of the other three patterns, which likely arose as a result of gene acquisition, deletion, and rearrangement events during the evolution of A. hydrophila, and may be linked to the acquisition of aerolysin genes. These findings shed light on the evolution of virulence in A. hydrophila.

Genotypic identification and study some virulence factors of Aeromonas hydrophila isolated from children's diarrheal specimens in Baghdad

In this study, 266 watery diarrhea specimens were collected from children less than 12 years that suffering from gastroenteritis in Welfare teaching hospital and Teaching laboratories/ Medical city. Out of 266 specimens, 19 (7.14) were identified as Aeromonas hydrophila isolates. The bacteria were identified as Aeromonas hydrophila phenotypically by Api-20 E system. Genotypic identification of Aeromonas hydrophila isolates also done by detection 16S rRNA gene by PCR assay and the results showed the presence of 16S rRNA gene in all 19 (100%) Aeromonas hydrophila studied isolates and this confirm the identification of Aeromonas hydrophila isolates. The antimicrobial susceptibility of Aeromonas hydrophila isolates was determined and the results showed that all 19 Aeromonas hydrophila isolates were resistant to Ampicillin and Amoxicillin (100%), while all 19 isolates were sensitive to Cefotaxime, Cefteriaxon, Norfloxacin and Imipenem (100%). Some virulence factors of the Aeromonas hydrophila isolates were investigated, all 19 Aeromonas hydrophila isolates were tested to detect their ability to produce hemolysin (toxin), protease, lipase, agglutinate human erythrocytes groups and ability to bind to Congo red. Out of 19 isolates, 14(73.68%) Aeromonas hydrophila isolates were hemolysin (toxin) producer which showed a clear zone of lysis on blood agar plates, 11(57.9%) isolates gave a positive result for protease production and 5(26.3%) isolates were gave a positive result for lipase production. Also isolates of Aeromonas hydrophila were tested for their ability to agglutinate human erythrocytes groups (O, A, B, AB). In which 10(52.6%) isolates showed agglutination with group (A) human erythrocytes, 9(47.4%) isolates showed agglutination with group (B) human erythrocytes, 9(47.4%) isolates showed agglutination with group (AB) human erythrocytes, 12(63.2%) isolates showed agglutination with group (O) human erythrocytes and 8 (42.1%) isolates showed agglutination with human erythrocytes groups (O, A, B, AB). The results of the ability to bind to Congo red revealed that 13(68.4%) isolates were bind to Congo red dye, while 6(31.6%) isolates were failed to bind to Congo red.

N-Acylhomoserine Lactones (AHLs), QseB/C Gene Detection, Virulence Factors and Antibiotics Resistance of <i>Aeromonas hydrophila</i>

The aim of this research was to detect the N-acyl homoserine lactones (AHLs) production and QseB/C gene of Aeromonas hydrophila. We analyzed the potentials of these isolates of Aeromonas hydrophila in causing biofilm formation, hemolysis, protease, and lipase. The antibiotic susceptibility of the 15 Aeromonas hydrophila isolates was also investigated. The detection of AHLs was carried out using the Chromobacterium violaceum strain CV026 as biosensors. The isolated strains were tested for the reaction of C. violaceum CV026 by cross-streaking on an agar plate. Production of AHLs was determined by the diffusing via the agar plates and the tinge of the biosensor strains. All isolated strains produced AHLs. A polymerase chain reaction (PCR) showed the isolated strains had qseB and qseC genes. Susceptibility tests of A. hydrophila isolates were administered against 25 different antibiotic disks representing 12 classes of antibiotics. The strains were highly resistant to β-Lactam with 96.7% showing resistibility, whereas 97.7% susceptibility was found towards Aminoglycoside class of the antibiotic used. 60% showed intermediate resistant to Polypeptide. 100% of the strains showed no resistant to Aminoglycoside, Polypeptide, Monobactam, and Carbapenems class of antibiotics. Each of the isolates was found to be associated with at least one virulent factor. Our results clearly demonstrated that there is a presence of QseB/C genes in A. hydrophila and also produces AHLs molecule and virulence factors. The investigated isolates showed the pathogenic potential of Aeromonas hydrophila which makes it a serious threat to public health.

The presence of plasmids in Aeromonas hydrophila and its relationship with antimicrobial and heavy metal-resistance profiles

ABSTRACT: Aeromonas hydrophila is a common fish pathogen that causes extensive damage to aquaculture. To develop and implement a more adequate strategy to farm fish, it is crucial to understand the bacterial-resistance levels and their transference dynamics. The objective of this study was to analyze the resistance profile of isolated Aeromonas hydrophila to antimicrobial agents and heavy metals and draw a correlation of the observed profiles with the presence of plasmids. Resistance of the isolated bacteria to antimicrobial agents (oxacilin, gentamicin, tetracycline, and nalidixic acid) and heavy metals (cadmium, lead, copper, and manganese) was verified using the minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) standards. The Multiple Antibiotic Resistance Index (MAR Index) was calculated. Plasmids were extracted by using a common methodology described elsewhere. Mann-Whitney Test, implemented in the R environment, was used to determine the correlation between resistance and plasmids presence. A high resistance to almost all antimicrobial agents and heavy metals was observed, except to gentamicin and cadmium. The MAR index results showed resistance to all antimicrobial profiles. Of the isolated bacteria, 14 showed the presence of plasmids. However, no correlation was noted between the resistance profile and the plasmid presence for these isolates, indicating that the genes responsible for resistance to microbial agents and heavy metals are present in the cromossomic DNA, which in turn suggested the possibility of gene transfer between the isolated bacteria. The resistance to heavy metals can be linked to heavy utilization of fertilizers along the Sao Francisco River.

The cytotoxicity effect of Metalloprotease produced and isolated from Aeromonas spp.

One hundred and fifty stool sampling were collected from clinical sources responsible of causing diarrhea in adults as well as children. The identification of Aeromonas hydrophila isolates depended on common methods of identification dependent on biochemical characteristics and culture, then vitek2 compact system was used. Eight Aeromonas hydrophila isolates were gained and revealed various productivity of metalloprotease; the isolate number 8 was the maximum effective in metalloprotease production. The eight isolates were examined with Polymerase Chain Reaction to prove enzyme gene presence, the results revealed that all isolates were positive for ahMP genes; metalloprotease was completely purified via a number of steps, which included ammonium sulphate precipitation, dialysis, ion exchange and gel chromatography. Cytotoxicity effect of metalloprotease studies on cancer and normal cell lines, The results showed the purified metalloprotease as effective cytotoxic effect on liver hepatocellular cancer cells (HepG2) compared with no effect on normal liver cell line (WRL-68) indicating less cytotoxic effect.

Aeromonas hydrophila from marketed mullet (Mugil cephalus) in Egypt: PCR characterization of β-lactam resistance and virulence genes.

Aeromonas hydrophila has been isolated from various fish species in Egypt and is known to carry virulence and antimicrobial resistance genes, which pose a risk for public health. The aim of the present study is to report, for the first time, the infection of mullet (Mugil cephalus) with A. hydrophila and to clarify the potential association between antimicrobial resistance and virulence traits encoded in A. hydrophila. In this study, the occurrence of A. hydrophila in marketed mullet and the antimicrobial resistance phenotypes of these isolates were determined. Aeromonas hydrophila isolates were screened for the presence of virulence and β-lactam resistance genes; the correlation between both gene groups was also investigated. The infection rate of examined mullet with A. hydrophila was 37% (50/135). The highest antimicrobial resistance was detected to cefoxitin (100%), followed by ampicillin (84%), ceftazidime (56%) and cefotaxime (40%). Only 4% of the isolates were resistant to erythromycin; 6% were resistant to both gentamicin and kanamycin with no resistance to ciprofloxacin. Variable frequencies of virulence and β-lactam resistance genes were evident from PCR, where aerA and blaTEM predominated. The study also indicated a general weak positive correlation (R=0·3) between both virulence and β-lactam resistance genes. Some of the studied virulence genes (e.g. aerA:hlyA and hlyA:ast) were found to correlate positively. The presence of virulence and resistance genes in A. hydrophila from food sources poses a serious threat to public health. To our knowledge, this is the first report describing the occurrence of A. hydrophila in mullet and highlighting the coexistence of virulence and β-lactam resistance genes encoded by these bacteria. These data provide insights into the potential association of antimicrobial resistance and virulence genes in A. hydrophila from marketed mullet in Egypt, which could pose threats to humans even if a weak positive correlation exists between both genes.