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Sample records for aeromonas hydrophila escherichia

  1. Cytotoxic activity of Aeromonas hydrophila.

    PubMed Central

    Donta, S T; Haddow, A D

    1978-01-01

    Most strains of Aeromonas hydrophila tested demonstrated cytotoxic activity on several tissue-cultured cell lines. The cytotoxin is heat-labile, non-dialyzable, and immunologically distinct from that of Shigella dysenteriae and Clostridium perfringens. None of the aeromonas isolates was found to be enterotoxigenic by either tissue culture or rabbit ileal loop assays. Images PMID:711344

  2. Molecular cloning and heterologous expression of laccase from Aeromonas hydrophila NIU01 in Escherichia coli with parameters optimization in production.

    PubMed

    Ng, I-Son; Zhang, Xia; Zhang, Yu; Lu, Yinghua

    2013-04-01

    Prior studies disclosed that Aeromonas hydrophila NIU01 was a biodecolorization and bioelectricity bacterium which was isolated from a cross-strait of Taiwan. However, enzymatic function, laccase, involved in this strain had never been reported. This first attempt is to explore its laccase activity, the molecular cloning and heterologous recombinant expression in Escherichia coli. A full-length novel gene of 1,647 bp, LacA, encoding of 549 amino acids was successfully cloned by polymerase chain reaction. The recombinant pET-15b(+)-NIU-LacA expression was compared in different E. coli strains. By applying Taguchi's L9 in culture optimization, the soluble laccase increased to 22.7 %, in which the conditions were obtained at 22 °C with initial shaking speed at 200 rpm, addition of lactose of 0.2 mM and CuSO4 of 0.5 mM to the medium, and shaking off while cell mass reached to OD(600nm) of 1.5. NIU-LacA was strongly inhibited by chloride ion. The optimal temperature was 60 °C and the optimum pH for ABTS (2,2'-azino-bis (3-ethylbenzthiazolinesulfonic acid) and 2,6-DMP (2,6-dimethoxyphenol) were pH 2.1 and pH 7.5 which enzymatic activity was 274.6 and 44.8 U/L, respectively. Further study in structural modeling of NIU-LacA showed the C terminal domain was the major variance in the three most closely A. hydrophila strains.

  3. Virulent Aeromonas hydrophila in channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study, we investigated factors that predisposed catfish to motile aeromonas septicemia (MAS) caused by virulent Aeromonas hydrophila (vAh). Our results revealed that wounding on fish body surface was a prerequisite for vAh infection and disease development. A reproducible waterborne challeng...

  4. Complete genome sequence of Aeromonas hydrophila AL06-06

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila occurs in freshwater environments and infects fish and mammals. In this work, we report the complete genome sequence of Aeromonas hydrophila AL06-06, which was isolated from diseased goldfish and is being used for comparative genomic studies with A. hydrophila strains causing ba...

  5. Aeromonas hydrophila Sepsis Mimicking Vibrio vulnificus Infection.

    PubMed

    Park, Se Young; Nam, Hyun Min; Park, Kun; Park, Seok Don

    2011-09-01

    Aeromonas hydrophila is a facultatively anaerobic, asporogenous gram-negative rod that has often been regarded as an opportunistic pathogen in hosts with impairment of a local or general defense mechanism. A 68-year-old alcoholic woman presented with shock and gangrene on the right arm. At first, her clinical presentations were severe painful erythematous swelling that worsened within a few hours with development of gangrene, edema, and blisters. Bullous fluid and blood cultures yielded A. hydrophila. Histopathological findings of sections obtained from the vesicle revealed subepidermal vesicles; necrosis of the epidermis, papillary dermis, and subcutaneous fat; and massive hemorrhage in the subcutis. Despite all efforts to save the patient, she died 8 hours after admission. Clinical features of A. hydrophila sepsis resemble those of Vibrio vulnificus sepsis. Therefore, in addition to the case report, we compared the cultural, biochemical, and morphological differences between A. hydrophila and V. vulnificus for facilitation of early and accurate identification of the causative agent.

  6. Aeromonas hydrophila Lateral Flagellar Gene Transcriptional Hierarchy

    PubMed Central

    Wilhelms, Markus; Gonzalez, Victor; Merino, Susana

    2013-01-01

    Aeromonas hydrophila AH-3 lateral flagella are not assembled when bacteria grow in liquid media; however, lateral flagellar genes are transcribed. Our results indicate that A. hydrophila lateral flagellar genes are transcribed at three levels (class I to III genes) and share some similarities with, but have many important differences from, genes of Vibrio parahaemolyticus. A. hydrophila lateral flagellum class I gene transcription is σ70 dependent, which is consistent with the fact that lateral flagellum is constitutively transcribed, in contrast to the characteristics of V. parahaemolyticus. The fact that multiple genes are included in class I highlights that lateral flagellar genes are less hierarchically transcribed than polar flagellum genes. The A. hydrophila lafK-fliEJL gene cluster (where the subscript L distinguishes genes for lateral flagella from those for polar flagella) is exclusively from class I and is in V. parahaemolyticus class I and II. Furthermore, the A. hydrophila flgAMNL cluster is not transcribed from the σ54/LafK-dependent promoter and does not contain class II genes. Here, we propose a gene transcriptional hierarchy for the A. hydrophila lateral flagella. PMID:23335410

  7. Draft genome sequence of Aeromonas hydrophila TN97-08

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is an opportunistic Gram-negative species causing disease in fish and mammals. The genus Aeromonas affects a variety of aquatic organisms and lives in diverse aquatic ecosystems (1). There are 39 A. hydrophila genomes currently available in GenBank. In the current study, we repo...

  8. DNA Adenine Methyltransferase Influences the Virulence of Aeromonas hydrophila

    PubMed Central

    Erova, Tatiana E.; Pillai, Lakshmi; Fadl, Amin A.; Sha, Jian; Wang, Shaofei; Galindo, Cristi L.; Chopra, Ashok K.

    2006-01-01

    Among the various virulence factors produced by Aeromonas hydrophila, a type II secretion system (T2SS)-secreted cytotoxic enterotoxin (Act) and the T3SS are crucial in the pathogenesis of Aeromonas-associated infections. Our laboratory molecularly characterized both Act and the T3SS from a diarrheal isolate, SSU of A. hydrophila, and defined the role of some regulatory genes in modulating the biological effects of Act. In this study, we cloned, sequenced, and expressed the DNA adenine methyltransferase gene of A. hydrophila SSU (damAhSSU) in a T7 promoter-based vector system using Escherichia coli ER2566 as a host strain, which could alter the virulence potential of A. hydrophila. Recombinant Dam, designated as M.AhySSUDam, was produced as a histidine-tagged fusion protein and purified from an E. coli cell lysate using nickel affinity chromatography. The purified Dam had methyltransferase activity, based on its ability to transfer a methyl group from S-adenosyl-l-methionine to N6-methyladenine-free lambda DNA and to protect methylated lambda DNA from digestion with DpnII but not against the DpnI restriction enzyme. The dam gene was essential for the viability of the bacterium, and overproduction of Dam in A. hydrophila SSU, using an arabinose-inducible, PBAD promoter-based system, reduced the virulence of this pathogen. Specifically, overproduction of M.AhySSUDam decreased the motility of the bacterium by 58%. Likewise, the T3SS-associated cytotoxicity, as measured by the release of lactate dehydrogenase enzyme in murine macrophages infected with the Dam-overproducing strain, was diminished by 55% compared to that of a control A. hydrophila SSU strain harboring the pBAD vector alone. On the contrary, cytotoxic and hemolytic activities associated with Act as well as the protease activity in the culture supernatant of a Dam-overproducing strain were increased by 10-, 3-, and 2.4-fold, respectively, compared to those of the control A. hydrophila SSU strain. The Dam

  9. [The occurrence of Escherichia coli, Aeromonas hydrophila, Plesiomonas shigelloides and Clostridium perfringens in the intestinal flora of gray herons (Ardea cinerea)].

    PubMed

    Glünder, G

    1989-05-01

    The flora of the large intestine of 92 grey herons was examined for the frequency of aerobic and microaerobic growing bacteria. Clostridium perfringens, Aeromonas hydrophila, Plesiomonas shigelloides and E. coli were isolated from 55%, 48%, 14% and 35% of the birds, respectively. It could be demonstrated that the findings of these bacteria in the intestinal flora are depending on the age of the birds. The percentage of carriers of Clostridium perfringens, Aeromonas hydrophila and Plesiomonas shigelloides was highest in nestlings younger than 18 days, less high in older nestlings and lowest in adult grey herons. Contrary to those bacteria, E. coli was found more often in the intestinal flora at increasing age of the birds. Salmonella spp. were isolated from 6 birds. Two birds yielded positive for Yersinia enterocolitica and Campylobacter spp., respectively. Other aerobic and microaerobic bacteria play a less significant role as part of the intestinal flora.

  10. Seasonal prevalence of nosocomial Aeromonas hydrophila infection related to aeromonas in hospital water.

    PubMed

    Picard, B; Goullet, P

    1987-09-01

    A seasonal variation in nosocomial Aeromonas hydrophila infection was correlated with the number of aeromonas in the hospital water supply. The high summer prevalence of A. hydrophila infection coincided with periods when water counts from storage tanks were highest. The waterborne origin of these infections highlights the importance of maintaining clean water supplies, especially where storage tanks are used. Monitoring A. hydrophila in hospital water, particularly during the summer months, may prove helpful.

  11. Function of a recombinant Chitinase derived from a virulent Aeromonas hydrophila isolated from diseased channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish (Ictalurus punctatus). Bioactive recombinant chitinase (rChi-Ah) was produced in Escherichia coli. Purified rChi-Ah had optimal activity at temperature of 42°C and pH 6.5. T...

  12. Genome Sequence of Hypervirulent Aeromonas hydrophila Strain HZAUAH

    PubMed Central

    Teng, Lin; Deng, Limei; Dong, Xingxing; Wei, Shun; Li, Jinquan

    2017-01-01

    ABSTRACT Aeromonas hydrophila, a zoonotic bacterium found in an expansive range of aquatic ecosystems, has been reported to cause severe diseases in fish, amphibians, reptiles, and mammals, including humans. Herein, we report the draft genome of the hypervirulent A. hydrophila strain HZAUAH isolated from a crucian in China. PMID:28302770

  13. Dynamics of Aeromonas hydrophila, Aeromonas sobria, and Aeromonas caviae in a sewage treatment pond.

    PubMed Central

    Monfort, P; Baleux, B

    1990-01-01

    The spatiotemporal dynamics of Aeromonas spp. and fecal coliforms in the sewage treatment ponds of an urban wastewater center were studied after 20 months of sampling from five stations in these ponds. Isolation and identification of 247 Aeromonas strains were undertaken over four seasons at the inflow and outflow of this pond system. The hemolytic activity of these strains was determined. The Aeromonas spp. and the fecal coliform distributions showed seasonal cycles, the amplitude of which increased at distances further from the wastewater source, so that in the last pond there was an inversion of the Aeromonas spp. cycle in comparison with that of fecal coliforms. The main patterns in these cycles occurred simultaneously at all stations, indicating control of these bacterial populations by seasonal factors (temperature, solar radiation, phytoplankton), the effects of which were different on each bacterial group. The analysis of the Aeromonas spp. population structure showed that, regardless of the season, Aeromonas caviae was the dominant species at the pond system inflow. However at the outflow the Aeromonas spp. population was dominated by A. caviae in winter, whereas Aeromonas sobria was the dominant species in the treated effluent from spring to fall. Among the Aeromonas hydrophila and A. sobria strains, 100% produced hemolysin; whereas among the A. caviae strains, 96% were nonhemolytic. Images PMID:2389929

  14. Aeromonas hydrophila: variability in biochemical characteristics of environmental isolates.

    PubMed

    Okpokwasili, G C

    1991-01-01

    A total of 161 strains, including 144 aeromonads, of which 100 had been isolated from water and sediment samples from the Chesapeake Bay, five clinical and 33 Anacostia River isolates of Aeromonas hydrophila, six Aeromonas sobria, and ten reference cultures were examined in a numerical taxonomy study in which each of the strains were examined for 56 unit characters. Similarity between strains was computed using the simple matching (SSM) and Jaccard (SJ) coefficients. The strains were clustered by unweighted average linkage and single linkage algorithms. Results of all analyses yielded similar clusterings of strains. Of the 161 strains included in the study, 148 were recovered in seven major clusters. A large cluster of Aeromonas hydrophila yielded two sub-clusters. From the results of the taxonomic analysis it is concluded that several biovars of Aeromonas hydrophila can be isolated from the aquatic environment, but geographical or site specific distribution of the biovars could not be detected.

  15. Pan-genome analysis of Aeromonas hydrophila, Aeromonas veronii and Aeromonas caviae indicates phylogenomic diversity and greater pathogenic potential for Aeromonas hydrophila.

    PubMed

    Ghatak, Sandeep; Blom, Jochen; Das, Samir; Sanjukta, Rajkumari; Puro, Kekungu; Mawlong, Michael; Shakuntala, Ingudam; Sen, Arnab; Goesmann, Alexander; Kumar, Ashok; Ngachan, S V

    2016-07-01

    Aeromonas species are important pathogens of fishes and aquatic animals capable of infecting humans and other animals via food. Due to the paucity of pan-genomic studies on aeromonads, the present study was undertaken to analyse the pan-genome of three clinically important Aeromonas species (A. hydrophila, A. veronii, A. caviae). Results of pan-genome analysis revealed an open pan-genome for all three species with pan-genome sizes of 9181, 7214 and 6884 genes for A. hydrophila, A. veronii and A. caviae, respectively. Core-genome: pan-genome ratio (RCP) indicated greater genomic diversity for A. hydrophila and interestingly RCP emerged as an effective indicator to gauge genomic diversity which could possibly be extended to other organisms too. Phylogenomic network analysis highlighted the influence of homologous recombination and lateral gene transfer in the evolution of Aeromonas spp. Prediction of virulence factors indicated no significant difference among the three species though analysis of pathogenic potential and acquired antimicrobial resistance genes revealed greater hazards from A. hydrophila. In conclusion, the present study highlighted the usefulness of whole genome analyses to infer evolutionary cues for Aeromonas species which indicated considerable phylogenomic diversity for A. hydrophila and hitherto unknown genomic evidence for pathogenic potential of A. hydrophila compared to A. veronii and A. caviae.

  16. Prevalence and distribution of Aeromonas hydrophila in the United States.

    PubMed Central

    Hazen, T C; Fliermans, C B; Hirsch, R P; Esch, G W

    1978-01-01

    The abundance of Aeromonas hydrophila was measured in 147 natural aquatic habitats in 30 states and Puerto Rico. Viable cell counts were used to estimate density at all sites by using Rimler-Shotts medium, a differential presumptive medium for A. hydrophila. Temperature, pH, conductivity, salinity, and turbidity were measured simultaneously with water sample collection. The density of A. hydrophila was higher in lotic than in lentic systems. Saline systems had higher densities of A. hydrophila than did freshwater systems. A. hydrophila could not be isolated from extremely saline, thermal, or polluted waters, even though it was found over wide ranges of salinity, conductivity, temperature, pH, and turbidity. Of the water quality parameters measured, only conductivity was significantly regressed with density of A. hydrophila. PMID:31839

  17. Experimental induction of motile Aeromonas septicemia in channel catfish by waterborne challenge with virulent Aeromonas hydrophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Motile Aeromonas septicemia (MAS), caused by virulent clonal isolates of Aeromonas hydrophila (vAh), is emerging as a major disease in catfish aquaculture in the Southeastern United States. Predisposing conditions leading to vAh infection in catfish were however largely unknown. The objective of thi...

  18. Infection and disease progress of motile Aeromonas septicemia caused by virulent Aeromonas hydrophila in channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Motile Aeromonas septicemia (MAS), caused by virulent clonal isolates of Aeromonas hydrophila (vAh), is emerging as a major disease in channel catfish (Ictalurus punctatus) aquaculture in the Southeastern United States. Predisposing conditions leading to vAh infection in catfish were however largely...

  19. Ciprofloxacin-resistant Aeromonas hydrophila cellulitis following leech therapy.

    PubMed

    Giltner, Carmen L; Bobenchik, April M; Uslan, Daniel Z; Deville, Jaime G; Humphries, Romney M

    2013-04-01

    We report a case of surgical site infection with ciprofloxacin-resistant Aeromonas hydrophila following leech therapy. Antimicrobial and genetic analyses of leech and patient isolates demonstrated that the resistant isolates originated from the leech gut microbiota. These data suggest that ciprofloxacin monotherapy as a prophylaxis regimen prior to leech therapy may not be effective in preventing infection.

  20. Ciprofloxacin-Resistant Aeromonas hydrophila Cellulitis following Leech Therapy

    PubMed Central

    Giltner, Carmen L.; Bobenchik, April M.; Uslan, Daniel Z.; Deville, Jaime G.

    2013-01-01

    We report a case of surgical site infection with ciprofloxacin-resistant Aeromonas hydrophila following leech therapy. Antimicrobial and genetic analyses of leech and patient isolates demonstrated that the resistant isolates originated from the leech gut microbiota. These data suggest that ciprofloxacin monotherapy as a prophylaxis regimen prior to leech therapy may not be effective in preventing infection. PMID:23363826

  1. Contribution of nuclease to the pathogenesis of Aeromonas hydrophila

    PubMed Central

    Ji, Yachan; Li, Jinquan; Qin, Zhendong; Li, Aihua; Gu, Zemao; Liu, Xiaoling; Lin, Li; Zhou, Yang

    2015-01-01

    Aeromonas hydrophila is a gram-negative bacterium that is widely distributed in aquatic environments and can cause septicemia in both fish and humans. However, the underlying mechanisms leading to severe infection are not well understood. In this study, an A. hydrophila nuclease (ahn) deletion mutant was constructed to investigate its contribution to pathogenesis. This mutant did not differ from the wild-type strain in terms of its growth or hemolytic phenotype. However, the ahn-deficient mutant was more susceptible to being killed by fish macrophages and mouse blood in vitro. Furthermore, evidence obtained using both fish and murine infection models strongly indicated that the inactivation of Ahn impaired the ability of A. hydrophila to evade innate immune clearance in vivo. More importantly, the virulence of the mutant was attenuated in both fish and mice, with reductions in dissemination capacities and mortality rates. These findings implicate Ahn in A. hydrophila virulence, with important functions in evading innate immune defenses. PMID:26039879

  2. Degradation of chitin and chitosan by a recombinant chitinase derived from a virulent Aeromonas hydrophila isolated from diseased channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish (Ictalurus punctatus). Bioactive recombinant chitinase (rChi-Ah) was produced in Escherichia coli. Purified rChi-Ah had optimal activity at temperature of 42°C and pH 6.5. T...

  3. Effects of cultural conditions on protease production by Aeromonas hydrophila.

    PubMed Central

    O'Reilly, T; Day, D F

    1983-01-01

    Production of extracellular proteolytic activity by Aeromonas hydrophila was influenced by temperature, pH, and aeration. Conditions which produced maximal growth also resulted in maximal protease production. Enzyme production appeared to be modulated by an inducer catabolite repression system whereby NH4+ and glucose repressed enzyme production and complex nitrogen and nonglucose, carbon energy sources promoted it. Under nutritional stress, protease production was high, despite poor growth. PMID:6342534

  4. Virulence and cytotoxicity of seafood borne Aeromonas hydrophila

    PubMed Central

    Illanchezian, Seethalakshmi; Jayaraman, SathishKumar; Manoharan, Muthu Saravanan; Valsalam, Saritha

    2010-01-01

    The present study was conducted to determine the virulence and cytotoxicity of Aeromonas hydrophila strains isolated from seafood samples collected from 5 major fish markets in Chennai, Tamil Nadu, India. Among 73 A. hydrophila strains isolated from fish and shrimp samples, 86.3% exhibited haemolysis, 78.1% produced slime, 98.63% produced protease and also demonstrated cytotoxicity on Vero cells. Cell shrinkage, detachment and rounding of Vero cells were recorded as cytotoxic changes. Only one strain did not show haemolysis, slime production, proteolytic activity and cytotoxicity on treatment with Vero cells. Positive correlation was observed between proteolytic activity and cytotoxicity irrespective of haemolytic activity of the strains. These results demonstrated the presence of wide spread, pathogenically characterized, cytotoxic seafood borne A. hydrophila in Chennai. PMID:24031577

  5. [Cloning and expression of a hemolysin gene of Aeromonas hydrophila and the immunogenicity of the toxoid].

    PubMed

    Zhang, Cuijuan; Yu, Zhouliang; Tian, Liying; Zhao, Baohua

    2009-02-01

    According to the GenBank sequences (GenBank Accession No. AF539467), one pair of primers was designed to amplify hly gene of Aeromonas hydrophila by PCR. After sequencing, homology analysis indicated that a DNA fragment of 1485 bp was amplified from isolated DNA from Aeromonas hydrophila, and it shared more than 99% homology in nucleotide sequence compared with other reference strains in GenBank. The gene was cloned in pET-28a vector to construct a recombinant plasmid pET-28a-hly, which was transformed into Escherichia coli BL21 (DE3), and the recombinant strain BL21(DE3)(pET-28a-hly) was obtained. The hemolysin was highly expressed when the recombinant strain BL21 (DE3) (pET-28a-hly) was induced by IPTG. The expressed protein was 56 kD as estimated by 15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the expressed Hly protein was confirmed by Western blotting. Mice were immunized with inactivated whole bacteria vaccine and the genetic engineering vaccines showing promise that all these vaccines have a high protective ability. The results showed that the recombinant strain BL21 (DE3)(pET-28a-hly) could be candidate of hemolysin toxoid vaccine to provide protective immunity against diseases caused by Aeromonas hydrophila.

  6. Virulence Factors of Aeromonas hydrophila: In the Wake of Reclassification

    PubMed Central

    Rasmussen-Ivey, Cody R.; Figueras, Maria J.; McGarey, Donald; Liles, Mark R.

    2016-01-01

    The ubiquitous “jack-of-all-trades,” Aeromonas hydrophila, is a freshwater, Gram-negative bacterial pathogen under revision in regard to its phylogenetic and functional affiliation with other aeromonads. While virulence factors are expectedly diverse across A. hydrophila strains and closely related species, our mechanistic knowledge of the vast majority of these factors is based on the molecular characterization of the strains A. hydrophila AH-3 and SSU, which were reclassified as A. piscicola AH-3 in 2009 and A. dhakensis SSU in 2013. Individually, these reclassifications raise important questions involving the applicability of previous research on A. hydrophila virulence mechanisms; however, this issue is exacerbated by a lack of genomic data on other research strains. Collectively, these changes represent a fundamental gap in the literature on A. hydrophila and confirm the necessity of biochemical, molecular, and morphological techniques in the classification of research strains that are used as a foundation for future research. This review revisits what is known about virulence in A. hydrophila and the feasibility of using comparative genomics in light of this phylogenetic revision. Conflicting data between virulence factors, secretion systems, quorum sensing, and their effect on A. hydrophila pathogenicity appears to be an artifact of inappropriate taxonomic comparisons and/or be due to the fact that these properties are strain-specific. This review audits emerging data on dominant virulence factors that are present in both A. dhakensis and A. hydrophila in order to synthesize existing data with the aim of locating where future research is needed. PMID:27610107

  7. Effect of copper sulfate on Aeromonas hydrophila infection in channel catfish (Ictalurus punctatus) fingerlings

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Motile Aeromonad Septicemia (MAS) results from primary or secondary infection with bacteria from Gram(-) Aeromonas spp., including Aeromonas hydrophila. Since 2009, an emerging strain of A. hydrophila has been associated, as a primary pathogen, with significant morbidity and mortality in the U.S. c...

  8. Whole-Genome Sequence of Aeromonas hydrophila Strain AH-1 (Serotype O11)

    PubMed Central

    Forn-Cuní, Gabriel; Tomás, Juan M.

    2016-01-01

    Aeromonas hydrophila is an emerging pathogen of aquatic and terrestrial animals, including humans. Here, we report the whole-genome sequence of the septicemic A. hydrophila AH-1 strain, belonging to the serotype O11, and the first mesophilic Aeromonas with surface layer (S-layer) to be sequenced. PMID:27587829

  9. Classification of a hypervirulent Aeromonas hydrophila pathotype responsible for epidemic outbreaks in warm-water fishes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the P...

  10. Aeromonas hydrophila and Aeromonas veronii Predominate among Potentially Pathogenic Ciprofloxacin- and Tetracycline-Resistant Aeromonas Isolates from Lake Erie

    PubMed Central

    Shinko, Jasmine; Augustyniak, Alexander; Gee, Christopher; Andraso, Greg

    2014-01-01

    Members of the genus Aeromonas are ubiquitous in nature and have increasingly been implicated in numerous diseases of humans and other animal taxa. Although some species of aeromonads are human pathogens, their presence, density, and relative abundance are rarely considered in assessing water quality. The objectives of this study were to identify Aeromonas species within Lake Erie, determine their antibiotic resistance patterns, and assess their potential pathogenicity. Aeromonas strains were isolated from Lake Erie water by use of Aeromonas selective agar with and without tetracycline and ciprofloxacin. All isolates were analyzed for hemolytic ability and cytotoxicity against human epithelial cells and were identified to the species level by using 16S rRNA gene restriction fragment length polymorphisms and phylogenetic analysis based on gyrB gene sequences. A molecular virulence profile was identified for each isolate, using multiplex PCR analysis of six virulence genes. We demonstrated that Aeromonas comprised 16% of all culturable bacteria from Lake Erie. Among 119 Aeromonas isolates, six species were identified, though only two species (Aeromonas hydrophila and A. veronii) predominated among tetracycline- and ciprofloxacin-resistant isolates. Additionally, both of these species demonstrated pathogenic phenotypes in vitro. Virulence gene profiles demonstrated a high prevalence of aerolysin and serine protease genes among A. hydrophila and A. veronii isolates, a genetic profile which corresponded with pathogenic phenotypes. Together, our findings demonstrate increased antibiotic resistance among potentially pathogenic strains of aeromonads, illustrating an emerging potential health concern. PMID:24242249

  11. Incidence and toxigenicity of Aeromonas hydrophila in seafood.

    PubMed

    Tsai, G J; Chen, T H

    1996-08-01

    Three selective media, Oxoid Aeromonas agar (OA), blood ampicillin agar (BA) and starch ampicillin agar (SA) were used to evaluate the presence of Aeromonas hydrophila in 66 samples of oyster, shrimp, fish and surimi products. Oyster had the highest incidence, with 50% positive, whilst no A. hydrophila was found in the surimi. Of the three selective media, BA displayed the highest recovery rate of A. hydrophila from seafood. Forty-eight isolates from this survey were tested for their capability to produce hemolysin and cytotoxin. Hemolysin was produced by 79.2% of the isolates and cytotoxin was produced by 91.7% of the isolates in brain heart infusion broth. One of the toxin-producing isolates from oyster, strain 8-169, was further tested for growth and toxin production in oyster, shrimp and fish at various temperatures. This particular isolate grew best and had highest toxin production in oyster. Hemolysin and cytotoxin were produced earlier at 28 degrees C than at 37 degrees C, and titers of hemolysin were also higher at 28 degrees C. At 5 degrees C, it was able to grow and produce hemolysin in oyster.

  12. Enterobacter asburiae and Aeromonas hydrophila: soft tissue infection requiring debridement.

    PubMed

    Koth, Kevin; Boniface, James; Chance, Elisha A; Hanes, Marina C

    2012-06-01

    Enterobacter asburiae and Aeromonas hydrophila are gram-negative bacilli that have been isolated in soil and water. Enterobacter asburiae can cause an array of diseases, and exposure to A hydrophila can cause soft tissue infections, including necrotizing faciitis.A healthy-appearing 22-year-old man presented with an innocuous soft tissue injury to his leg due to an all-terrain vehicle crash. He received intravenous antibiotics and was discharged with prophylactic oral antibiotics. After the rapid onset of high fevers (102°F-103°F) <24 hours postinjury, he returned to the emergency department. Emergent surgical debridement was performed, and broad-spectrum intravenous antibiotics were started. Fevers persisted, and the patient underwent repeat extensive surgical debridement and antibiotic bead placement <30 hours after the initial surgical debridement and broad-spectrum antibiotics. Intraoperative cultures found E asburiae and A hydrophila in the wound. Following a long course of antibiotics and a skin graft, he fully recovered and had no functional deficits 1 year postoperatively.Extensive research revealed that these organisms are rare in soft tissue infections. E asburiae is opportunistic but has not been reported as a primary wound organism, and A hydrophila infections have been reported following motor vehicle crashes involving wound contamination. At presentation, it is challenging to determine rare organisms in a timely fashion; however, emergent extensive surgical intervention of an accelerated aberrant disease process should be considered to avoid catastrophic outcomes.

  13. Cloning, Sequencing, and Role in Serum Susceptibility of Porin II from Mesophilic Aeromonas hydrophila

    PubMed Central

    Nogueras, Maria Mercé; Merino, Susana; Aguilar, Alicia; Benedi, Vicente Javier; Tomás, Juan M.

    2000-01-01

    We cloned and sequenced the structural gene for Aeromonas hydrophila porin II from strain AH-3 (serogroup O:34). The genetic position of this gene, like that of ompF in Escherichia coli, is adjacent to aspC and transcribed in the same direction. However, upstream of the porin II gene no similarities with E. coli were found. We obtained defined insertion mutants in porin II gene either in A. hydrophila (O:34) or A. veronii sobria (serogroup O:11) serum-resistant or -sensitive strains. Furthermore, we complemented these mutants with a plasmid harboring only the porin II gene, which allowed us to define the role of porin II as an important surface molecule involved in serum susceptibility and C1q binding in these strains. PMID:10722573

  14. Novel Aeromonas hydrophila PPD134/91 Genes Involved in O-Antigen and Capsule Biosynthesis

    PubMed Central

    Zhang, Y. L.; Arakawa, E.; Leung, K. Y.

    2002-01-01

    The sequences of the O-antigen and capsule gene clusters of the virulent Aeromonas hydrophila strain PPD134/91 were determined. The O-antigen gene cluster is 17,296 bp long and comprises 17 genes. Seven pathway genes for the synthesis of rhamnose and mannose, six transferase genes, one O unit flippase gene, and one O-antigen chain length determinant gene were identified by amino acid sequence similarity. PCR and Southern blot analysis were performed to survey the distribution of these 17 genes among 11 A. hydrophila strains of different serotypes. A. hydrophila PPD134/91 might belong to serotype O:18, as represented by JCM3980; it contained all the same O-antigen genes as JCM3980 (97 to 100% similarity at the DNA and amino acid levels). The capsule gene cluster of A. hydrophila PPD134/91 is 17,562 bp long and includes 13 genes, which were assembled into three distinct regions similar to those of the group II capsule gene cluster of Escherichia coli and other bacteria. Regions I and III contained four and two capsule transport genes, respectively. Region II had five genes which were highly similar to capsule synthesis pathway genes found in other bacteria. Both the purified O-antigen and capsular polysaccharides increased the ability of the avirulent A. hydrophila strain PPD35/85 to survive in naïve tilapia serum. However, the purified surface polysaccharides had no inhibitory effect on the adhesion of A. hydrophila PPD134/91 to carp epithelial cells. PMID:11953367

  15. Draft genome sequences of four virulent aeromonas hydrophila strains from catfish aquaculture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since 2009, a clonal group of virulent Aeromonas hydrophila (VAh) strains has been causing severe disease in the catfish aquaculture industry in the Southeastern United States. Here, we report draft genomes of four A. hydrophila isolates from catfish aquaculture that represent this clonal group....

  16. Over-expression, purification and immune responses to Aeromonas hydrophila AL09-73 flagellar proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is ubiquitous in aquatic environments worldwide and causes many diseases in fish as well as human. Recent outbreaks of aeromonad diseases in channel catfish prompted us to investigate catfish immune responses during infection of A. hydrophila. In this communication, we report ...

  17. Draft Genome Sequences of Four Virulent Aeromonas hydrophila Strains from Catfish Aquaculture

    PubMed Central

    Tekedar, Hasan C.; Kumru, Salih; Karsi, Attila; Waldbieser, Geoffrey C.; Sonstegard, Tad; Schroeder, Steven G.; Liles, Mark R.; Griffin, Matt J.

    2016-01-01

    Since 2009, a clonal group of virulent Aeromonas hydrophila strains has been causing severe disease in the catfish aquaculture industry in the southeastern United States. Here, we report draft genomes of four A. hydrophila isolates from catfish aquaculture that represent this clonal group. PMID:27540076

  18. Draft Genome Sequences of Three Aeromonas hydrophila Isolates from Catfish and Tilapia

    PubMed Central

    Tekedar, Hasan C.; Kumru, Salih; Kalindamar, Safak; Karsi, Attila; Waldbieser, Geoffrey C.; Sonstegard, Tad; Schroeder, Steven G.; Liles, Mark R.; Griffin, Matt J.

    2017-01-01

    ABSTRACT Aeromonas hydrophila is a Gram-negative bacterium that is particularly adapted to freshwater environments and can cause severe infections in fish and humans. Here, we report the draft genomes of three A. hydrophila catfish and tilapia isolates. PMID:28104665

  19. Genome Sequence of Aeromonas hydrophila Strain AH-3 (Serotype O34)

    PubMed Central

    Forn-Cuní, Gabriel; Tomás, Juan M.

    2016-01-01

    Aeromonas hydrophila is an emerging pathogen of poikilothermic animals, from fish to mammals, including humans. Here, we report the whole-genome sequence of the A. hydrophila AH-3 strain, isolated from a fish farm goldfish septicemia outbreak in Spain, with a characterized polar and lateral flagellum glycosylation pattern. PMID:27587828

  20. Cold Shock Exoribonuclease R(VacB) is involved in Aeromonas hydrophila Virulence

    EPA Science Inventory

    In this study, we cloned and sequenced a virulence-associated gene (vacB) from a clinical isolate SSU of Aeromonas hydrophila. We identified this gene based on our recently annotated genome sequence of the environmental isolate ATCC 7966T of A. hydrophila and the vacB gene of Shi...

  1. Cold Shock Exoribonuclease R (VacB) is Involved in Aeromonas hydrophila Pathogenesis

    EPA Science Inventory

    In this study, we cloned and sequenced a virulence-associated gene (vacB) from a clinical isolate SSU of Aeromonas hydrophila. We identified this gene based on our recently annotated genome sequence of the environmental isolate ATCC 7966T of A. hydrophila and the vacB gene of Shi...

  2. Aeromonas hydrophila flagella glycosylation: involvement of a lipid carrier.

    PubMed

    Merino, Susana; Fulton, Kelly M; Twine, Susan M; Wilhelms, Markus; Molero, Raquel; Tomás, Juan M

    2014-01-01

    Polar flagellin proteins from Aeromonas hydrophila strain AH-3 (serotype O34) were found to be O-glycosylated with a heterogeneous glycan. Mutants unable to produce WecP or Gne enzymes showed altered motility, and the study of their polar flagellin glycosylation showed that the patterns of glycosylation differed from that observed with wild type polar flagellin. This suggested the involvement of a lipid carrier in glycosylation. A gene coding for an enzyme linking sugar to a lipid carrier was identified in strain AH-3 (WecX) and subsequent mutation abolished completely motility, flagella production by EM, and flagellin glycosylation. This is the first report of a lipid carrier involved in flagella O-glycosylation. A molecular model has been proposed. The results obtained suggested that the N-acetylhexosamines are N-acetylgalactosamines and that the heptasaccharide is completely independent of the O34-antigen lipopolysaccharide. Furthermore, by comparing the mutants with differing degrees of polar flagellin glycosylation, we established their importance in A. hydrophila flagella formation and motility.

  3. Physiological control of amonabactin biosynthesis in Aeromonas hydrophila.

    PubMed

    Barghouthi, S; Young, R; Arceneaux, J E; Byers, B R

    1989-01-01

    Amonabactin is a siderophore from Aeromonas hydrophila which is produced in two biologically active forms composed of the phenolate 2,3-dihydroxybenzoic acid (DHB), lysine, glycine, and either trytophan (amonabactin T) or phenylalanine (amonabactin P). Amonabactin biosynthetic mutants (generated by chemical mutagenesis) that either produced no amonabactin or overproduced the siderophore were isolated and identified on chrome azurol S siderophore detection agar. Amonabactin-negative mutants were of two categories. One type produced no phenolates and used exogenous DHB to synthesize amonabactin (both forms) while the other type excreted DHB but not amonabactin. This suggests an amonabactin biosynthetic pathway composed of two segments, one producing DHB and the other assembling amonabactin from DHB and the amino acids. Overproduction mutants used amonabactin poorly or not at all, indicating that they contained lesions in amonabactin utilization. Adding the analog D-tryptophan to wild-type A. hydrophila cultures reduced synthesis of both amonabactin T and amonabactin P and lengthened the lag phase in iron restricted medium. The tryptophan and phenylalanine forms of amonabactin may be synthesized by a single assembly pathway that contains a novel enzyme (sensitive to D-tryptophan) which inserts either tryptophan or phenylalanine into amonabactin.

  4. Carbohydrate-reactive, pore-forming outer membrane proteins of Aeromonas hydrophila.

    PubMed Central

    Quinn, D M; Atkinson, H M; Bretag, A H; Tester, M; Trust, T J; Wong, C Y; Flower, R L

    1994-01-01

    Two outer membrane proteins of Aeromonas hydrophila A6, isolated in a one-step affinity chromatography process based on carbohydrate reactivity, were found to be pore-forming molecules in artificial planar bilayer membranes. These carbohydrate-reactive outer membrane proteins (CROMPs; M(r)s, 40,000 and 43,000) were subjected to amino acid analysis. The amino acid profiles for these two outer membrane proteins were almost identical. A partial protein sequence of a 14-amino-acid fragment of the 40,000-Da protein revealed homology with outer membrane porins of Escherichia coli and A. hydrophila. CROMPs were compared with carbohydrate-reactive porins also extracted from outer membranes of A. hydrophila A6. These porins were isolated by using standard porin purification techniques (insolubility in 2% sodium dodecyl sulfate, solubility in 0.4 M NaCl, and Sephacryl S-200 gel filtration), and then Synsorb H type 2 affinity chromatography was done. The physical and functional properties of the carbohydrate-reactive porins and CROMPs were found to be identical. On the basis of pore-forming properties in planar lipid bilayers and channel inhibition with maltotriose solutions, a nonspecific, general diffusion porin and a LamB-like maltoporin were identified in both CROMP and carbohydrate-reactive porin preparations. To our knowledge, the use of carbohydrate reactivity to isolate channel-forming proteins from bacterial outer membranes has not been reported previously. Images PMID:7520425

  5. In vitro comparisons of the inhibitory activity of florfenicol copper sulfate and potassium permanganate towards Aeromonas hydrophila and Flavobacterium columnare

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila and Flavobacterium columnare, the etiological agents of motile aeromonas septicemia (MAS) and columnaris disease, respectively, have been recently causing crippling moralities to the sunshine bass, Morone chrysops female X Morone saxatilis male (Percichthyidae), industry in the ...

  6. Comparative genomics of Aeromonas hydrophila isolates from an epidemic in channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background Aeromonas hydrophila was identified as the etiologic agent infecting farmed channel catfish in 2009/2010, resulting in higher mortality rates than typical for motile Aeromonas septicemia with over 5 million pounds of catfish lost to this outbreak. The biochemistry, molecular phylogeny, an...

  7. Chitin degradation and utilization by virulent Aeromonas hydrophila strain ML10-51K

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Virulent Aeromonas hydrophila (vAh) is one of the most important bacterial pathogens that causes persistent outbreaks of motile Aeromonas septicemia (MAS) in warm-water fishes. Among factors associated with MAS outbreaks, the survivability of this pathogen in aquatic environments is of great concern...

  8. [Incidence, behavior and control of Aeromonas hydrophila in meat and dairy products].

    PubMed

    García-López, M L; Otero, A; García-Fernández, M C; Santos, J A

    1993-02-01

    This review deals with several aspects of Aeromonas hydrophila and other motile Aeromonas species associated with foodborne illness. Although it is mainly dedicated to the factors affecting growth and survival of this species in foods of animal origin, information on other topics is also provided. This paper includes sections on: Taxonomy, diseases caused by Aeromonas, virulence factors, reservoirs and prevalence in foods and water, factors affecting growth and survival, isolation and identification, and control measures.

  9. Potential of Psidium guajava supplemented fish diets in controlling Aeromonas hydrophila infection in tilapia (Oreochromis niloticus).

    PubMed

    Pachanawan, Adithepchaikarn; Phumkhachorn, Parichat; Rattanachaikunsopon, Pongsak

    2008-11-01

    Fourteen herbs were extracted in water, 95% ethanol, and ether, and tested in vitro for antimicrobial activities against Aeromonas hydrophila, a fish pathogen causing motile Aeromonas septicemia. Using swab paper disc assays and minimal inhibitory concentration (MIC) determinations, we noted that the ethanol extract of Psidium guajava leaf exhibited the strongest antimicrobial activity. The extract allowed growth-inhibited A. hydrophila cells to regrow in fresh BHI broth indicating a bacteriostatic mode of action. In a pathogenicity test, the median lethal dose (LD(50)) of A. hydrophila for tilapia (Oreochromis niloticus) by intraperitoneal injection was 3.44 x 10(6) CFU/ml. In vivo experiments showed that fish diets containing either dry leaf powder of P. guajava or dried ethanol extract of P. guajava leaf reduced mortality of A. hydrophila infected tilapia with no detected adverse effect on the fish. This study suggests that P. guajava leaf has the potential to control fish diseases caused by A. hydrophila.

  10. Implication of lateral genetic transfer in the emergence of Aeromonas hydrophila isolates of epidemic outbreaks in channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: A recent epidemic outbreak of motile Aeromonas septicemia of catfish caused by highly virulent Aeromonas hydrophila is a major threat to the catfish industry in the southeastern United States. The lack of a complete genome sequence for this newly emerged A. hydrophila genotype hampers ef...

  11. Construction, expression and characterization of eleven putative flagellar apparatus genes of Aeromonas hydrophila AL09-73

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is ubiquitous in aquatic environments worldwide and is responsible for many human and fish diseases. The genome sequence of A. hydrophila is available in the GenBank database. Our ultimate goal is to develop the whole genome protein arrays of A. hydrophila. In this study, we ...

  12. The impact of quorum sensing on the virulence of Aeromonas hydrophila and Aeromonas salmonicida towards burbot (Lota lota L.) larvae.

    PubMed

    Natrah, F M I; Alam, Md Iftakharul; Pawar, Sushant; Harzevili, A Shiri; Nevejan, Nancy; Boon, Nico; Sorgeloos, Patrick; Bossier, Peter; Defoirdt, Tom

    2012-09-14

    In this study, the link between quorum sensing in Aeromonas spp. and its virulence towards burbot (Lota lota) was investigated. High mortality occurred in burbot juveniles challenged with Aeromonas salmonicida HN-00, but not in juveniles challenged with Aeromonas hydrophila AH-1N. Meanwhile, both A. hydrophila AH-1N and A. salmonicida HN-00 were virulent towards larvae. The effect of quorum sensing on the virulence of A. hydrophila AH-1N towards burbot larvae was further investigated using quorum sensing mutants (N-(butyryl)-L-homoserine lactone production and receptor mutants). Challenge with these mutants resulted in higher survival of burbot larvae when compared to challenge with the wild type, and the addition of the signal molecule N-butyryl-L-homoserine lactone restored the virulence of the quorum sensing production mutant. Moreover, quorum sensing inhibitors protected the burbot larvae from both Aeromonas strains. Finally, the freshwater micro-algae Chlorella saccharophila and Chlamydomonas reinhardtii, which are able to interfere with quorum sensing, also protected burbot from the pathogens. However, QS interference was unlikely to be the only mechanism. This study revealed that the virulence of Aeromonas spp. towards burbot is regulated by quorum sensing and that quorum sensing inhibitors and micro-algae are promising biocontrol agents.

  13. Biological approach to synthesize TiO2 nanoparticles using Aeromonas hydrophila and its antibacterial activity

    NASA Astrophysics Data System (ADS)

    Jayaseelan, Chidambaram; Rahuman, Abdul Abdul; Roopan, Selvaraj Mohana; Kirthi, Arivarasan Vishnu; Venkatesan, Jayachandran; Kim, Se-Kwon; Iyappan, Moorthy; Siva, Chinnadurai

    2013-04-01

    Nanosized materials have been an important subject in basic and applied sciences. A novel, low-cost, green and reproducible bacteria, Aeromonas hydrophila mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs) was reported. The resulting nanoparticles were characterized by FTIR, XRD, AFM and FESEM with EDX. FTIR showed characteristic bands (1643 and 3430 cm-1) finds the role of carboxyl group Osbnd H stretching amine Nsbnd H stretch in the formation of TiO2 NPs. The XRD spectrum confirmed that the synthesized TiO2 NPs were in the form of nanocrystals, as evidenced by the peaks at 2θ values of 27.47°, 31.77°, 36.11°, 41.25°, 54.39°, 56.64° and 69.54° were identified as 110, 100, 101, 111, 211, 220 and 301 reflections, respectively. The crystallite sizes were calculated using Scherrer's formula applied to the major intense peaks and found to be the size of 40.50 nm. The morphological characterization was analyzed by FESEM and the analysis showed the NPs smooth shaped, spherical and uneven. GC-MS analysis showed the main compounds found in A. hydrophila were uric acid (2.95%), glycyl-L-glutamic acid (6.90%), glycyl-L-proline (74.41%) and l-Leucyl-d-leucine (15.74%). The potential glycyl-L-proline could have played an important role as a capping agent. A possible mechanism for the biosynthesis of TiO2 NPs has been proposed. The antibacterial activity of the synthesized TiO2 NPs was assessed by well diffusion method toward A. hydrophila, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes and Enterococcus faecalis and showed effective inhibitory activity against S. aureus (33 mm) and S. pyogenes (31 mm).

  14. Role of MshQ in MSHA pili biosynthesis and biofilm formation of Aeromonas hydrophila.

    PubMed

    Qin, Y X; Yan, Q P; Mao, X X; Chen, Z; Su, Y Q

    2014-10-31

    Biofilm formation of pathogen bacterium is currently one of the most widely studied topics; however, little is known regarding pathogen bacteria biofilms in aquaculture. Aeromonas hydrophila is a representative species of the genus Aeromonas, which has been recognized as a common pathogen, is associated with many diseases in aquatic animals, and causes significant mortality. The objectives of this study are i) to confirm that A. hydrophila can form biofilms on abiotic substrates and construct a biofilm growth curve for this bacterium; ii) to identify the genes that play crucial roles in A. hydrophila biofilm formation. The biofilm growth curve of A. hydrophila was constructed using a crystal violet assay, which showed that biofilm formation for this bacterium is a dynamic process. Next, a mutant library of pathogenic A. hydrophila B11 was constructed using the mini-Tn10 transposon mutagenesis system. A total of 861 mutants were screened, and 5 mutants were stably deficient in biofilm formation. Molecular analysis of the mutant B112 revealed that the open reading frame that encodes the protein MshQ was disrupted. Comparison of biological characteristics including growth, motility, and adhesion between the mutant B112 and the wild-type strain B11 suggested that MshQ is necessary for mannose-sensitive hemagglutinin pilus biosynthesis of A. hydrophila, and that these pili play crucial roles in A.hydrophila adherence to a solid surface during the early stages of biofilm formation.

  15. Survival and recovery of Aeromonas hydrophila in water: development of methodology for testing bottled water in Canada.

    PubMed

    Warburton, D W; McCormick, J K; Bowen, B

    1994-02-01

    Proposed changes to the Regulations for bottled water in the Food and Drugs Act of Canada include criteria for Aeromonas hydrophila (0 colony-forming units/100 mL water). The development of the methodology used to support these proposed Regulations and the survival of A. hydrophila in inoculated water are described. The methodology used in the isolation of A. hydrophila includes the use of hydrophobic grid membrane filters (HGMF), a resuscitation step on tryptic soy agar, and selective plating on membrane-Aeromonas-trehalose agar and Aeromonas medium. Aeromonas hydrophila proliferated and survived in inoculated water for up to 60 days or longer depending on the other contaminating bacteria. The presence of Pseudomonas aeruginosa enhanced the survival of A. hydrophila and enabled this bacteria to survive for more than 60 days.

  16. Bacteriocin-like substance (BLS) production in Aeromonas hydrophila water isolates.

    PubMed

    Messi, Patrizia; Guerrieri, Elisa; Bondi, Moreno

    2003-03-14

    30 Aeromonas hydrophila water isolates were tested for bacteriocin-like substance (BLS) production using a target panel of closely related microorganisms and other Gram-positive and Gram-negative bacteria, including food-borne pathogens. A. hydrophila showed antibacterial activity against one or more indicator microorganisms, but the activity emerged only with non-phylogenetically related genera or species. In particular all A. hydrophila showed antibacterial activity against one or more of the tested Staphylococcus strains, five against Listeria spp. (Listeria seeligeri, Listeria welshimeri and Listeria ivanovii), and eight presented a weak antagonistic activity towards Streptococcus agalactiae and Lactobacillus spp. Inhibitory activity was not observed against the other Gram-positive (Listeria monocytogenes, Listeria innocua and Enterococcus spp.) and Gram-negative tested strains, including Aeromonas sobria, Aeromonas caviae and the same A. hydrophila, when used as indicator. Anti-staphylococcal activity was observed with a gradual increase of the inhibition zone during incubation and seemed to be influenced by A. hydrophila hemolytic expression. Extrachromosomal analysis showed the presence, in 70% of the strains, of one to five plasmids with molecular masses ranging from 2.1 to 41.5 MDa, but it was not possible to relate this result with BLS production.

  17. Rapid quantitative detection of Aeromonas hydrophila strains associated with disease outbreaks in catfish aquaculture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the summer of 2009, a new strain of Aeromonas hydrophila was implicated in severe disease outbreaks in farm-raised catfish in Alabama, Arkansas and Mississippi. These outbreaks mostly afflicted large fish and resulted in considerable losses in short periods. Given the rapid onset and biosecurity ...

  18. Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila

    EPA Science Inventory

    Our laboratory recently molecularly characterized the type II secretion system (T2SS)-associated cytotoxic enterotoxin (Act) and the T3SS-secreted AexU effector from a diarrheal isolate SSU of Aeromonas hydrophila. The role of these toxin proteins in the pathogenesis of A. hydrop...

  19. Flagellar apparatus gene sequences of Aeromonas hydrophila AL09-73 isolate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Flagellar apparatus genes of recent outbreak Aeromonas hydrophila AL09-73 isolate were sequenced and characterized. Total 28 flagellar genes were identified. The sizes of the genes range from 318 to 2001 nucleotides, which potentially encode different complex flagellar proteins. At nucleotide and...

  20. Survival of Aeromonas hydrophila and Listeria monocytogenes on fresh vegetables stored under moderate vacuum.

    PubMed

    Aytac, S A; Gorris, L G

    1994-11-01

    Storage at 6.5°C under moderate vacuum effectively prevented growth of Aeromonas hydrophila on chicory endive, but had only a limited inhibitory effect on the growth of the organism on mung bean sprouts. Growth of Listeria monocytogenes on chicory endive was strongly stimulated under these conditions, whereas it was decreased on mung-bean sprouts.

  1. [Aeromonas hydrophila in the drinking water in Djibouti: commensal germ or diarrhea-causing agent?].

    PubMed

    Fox, E; Mikhail, I A; Haberberger, R L; Abbatte, E A; Ahmed, M H

    1990-04-01

    To investigate the bacteriological quality of drinking water used by inhabitants of the Republic of Djibouti who were not supplied with piped running water, we analysed 16 fresh-water samples from various sources. Only 3 samples were sterile; they were taken from village pumps and from a water-truck. Eleven samples yielded colonies of Aeromonas hydrophila too numerous to be counted; they were taken from water tanks, metal barrels, or wells dug in either dry river beds or along the seashore. We speculate that this high isolation frequency of Aeromonas hydrophila in fresh water samples may be related to conditions that are exceptionally favourable for the growth of the bacterium (e.g. high temperature and elevated concentrations of certain salts and minerals in the fresh water of Djibouti). We wonder nevertheless whether the infected water supplies were a source of diarrhoea for humans. Indeed, antibiotic resistance patterns were dissimilar when the 11 environmental strains were compared to 7 strains of Aeromonas hydrophila isolated from diarrhoeal patients in Djibouti during the same period. More studies are needed to determine if Aeromonas hydrophila is always a commensal inhabitant of fresh water in Djibouti, or if it can be a cause of infectious diarrhoea. Accordingly, Public Health authorities in Djibouti will be able to decide if water from wells and tanks is safe for drinking, or if it needs disinfection before consumption.

  2. Detection and quantification of virulent Aeromonas hydrophila in channel catfish tissues following waterborne challenge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this study was to understand the pathogenesis of motile aeromonas septicemia caused by virulent A. hydrophila (vAh) in channel catfish, Ictalurus punctatus. Adipose fin clipped catfish were challenged with vAh using waterborne challenge method and the distribution of vAh in catfish tissue...

  3. Immunization with recombinant aerolysin and hemolysin protected channel catfish against virulent Aeromonas hydrophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is emerging as one of the major concerns in catfish aquaculture in the Southeastern United States due to recent outbreaks of motile aeromonad septicemia (MAS) caused by virulent clonal isolates. There is no effective vaccine currently available for the prevention of MAS. In this...

  4. Humoral immune response of catfish immunized with extracellular products of Aeromonas hydrophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is emerging as one of the major concerns in catfish aquaculture in the Southeastern United States due to recent outbreaks of motile aeromonad septicemia (MAS) caused by the pathogen. Prophylactic treatment is being sought to prevent MAS. Since multiple virulence associated facto...

  5. Diagnosis of Aeromonas hydrophila, Mycobacterium species, and Batrachochytrium dendrobatidis in an African Clawed Frog (Xenopus laevis)

    PubMed Central

    Hill, William A; Newman, Shelley J; Craig, Linden; Carter, Christopher; Czarra, Jane; Brown, J Paige

    2010-01-01

    Here we describe diagnosis of concurrent infection with Aeromonas hydrophila, Mycobacterium spp., and Batrachochytrium dendrobatidis in a wild female Xenopus laevis captured in Chile and transported to the United States. After approximately 130 d in the laboratory, the frog was presented for dysecdysis and obtundation. After euthanasia, tissues were submitted for histopathologic evaluation and PCR analysis for B. dendrobatidis and Ranavirus. Clinically significant gross lesions included cutaneous ulcerations on the lip, right forelimb, and ventral chest. Microscopic findings included regionally extensive splenic necrosis, diffuse pneumonia, and fibrinous coelomitis all containing intralesional bacteria. PCR analysis yielded positive results for B. dendrobatidis only. Bacterial culture of the ulcerated skin and liver yielded A. hydrophila. Infection with Contracaecum spp. was diagnosed as an incidental finding. To our knowledge, this case is the first report of simultaneous infection with Aeromonas hydrophila, Mycobacterium spp., and Batrachochytrium dendrobatidis in a laboratory-maintained X. laevis captured from the wild. PMID:20353698

  6. Experimental challenge studies in Vietnamese catfish, Pangasianodon hypophthalmus (Sauvage), exposed to Edwardsiella ictaluri and Aeromonas hydrophila.

    PubMed

    Crumlish, M; Thanh, P C; Koesling, J; Tung, V T; Gravningen, K

    2010-09-01

    The two main diseases in the pangasius catfish industry are bacillary necrosis of Pangasianodon (BNP) and motile aeromonas septicaemia (MAS), where the aetiological agents have been identified as Edwardsiella ictaluri and Aeromonas hydrophila, respectively. In this study, apparently healthy Pangasianodon hypophthalmus were exposed to E. ictaluri, A. hydrophila or both bacterial species by intraperitoneal injection or immersion. There were 20 fish per treatment group, and the bacterial isolates used for the study were recovered from natural infections of BNP or MAS in farmed Vietnamese P. hypophthalmus. The results of the experimental infections mimicked the natural disease outbreaks reported from these pathogens in P. hypophthalmus. Furthermore, it was clearly demonstrated that E. ictaluri was only recovered from the fish exposed to the bacterium and not recovered from the animals receiving A. hydrophila.

  7. Pathogenicity of Aeromonas hydrophila, Klebsiella pneumoniae, and Proteus mirabilis to brown tree frogs (Litoria ewingii).

    PubMed

    Schadich, Ermin; Cole, Anthony L J

    2010-04-01

    Bacterial dermatosepticemia, a systemic infectious bacterial disease of frogs, can be caused by several opportunistic gram-negative bacterial species including Aeromonas hydrophila, Chryseobacterium indologenes, Chryseobacterium meningosepticum, Citrobacter freundii, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and Serratia liquifaciens. Here we determined the pathogenicity of 3 bacterial species (Aeromonas hydrophila, Klebsiella pneumoniae, and Proteus mirabilis) associated with an outbreak of fatal dermatosepticemia in New Zealand Litoria ewingii frogs. A bath challenge method was used to expose test frogs to individual bacterial species (2 x 10(7) cfu/mL in pond water); control frogs were exposed to uninfected pond water. None of the control frogs or those exposed to A. hydrophila or P. mirabilis showed any morbidity or mortality. Morbidity and mortality was 40% among frogs exposed to K. pneumonia, and the organism was reisolated from the hearts, spleens, and livers of affected animals.

  8. Aeromonas hydrophila subsp. dhakensis Isolated from Feces, Water and Fish in Mediterranean Spain

    PubMed Central

    Esteve, Consuelo; Alcaide, Elena; Blasco, María Dolores

    2012-01-01

    Eight Aeromonas hydrophila-like arabinose-negative isolates from diverse sources (i.e., river freshwater, cooling-system water pond, diseased wild European eels, and human stools) sampled in Valencia (Spain) during 2004–2005, were characterized by 16S rRNA gene sequencing and extensive biochemical testing along with reference strains of most Aeromonas species. These isolates and all reference strains of A. hydrophila subsp. dhakensis and A. aquariorum showed a 16S rRNA sequence similarity of 99.8–100%, and they all shared an identical phenotype. This matched exactly with that of A. hydrophila subsp. dhakensis since all strains displayed positive responses to the Voges-Prokauer test and to the use of dl-lactate. This is the first report of A. hydrophila subsp. dhakensis recovered from environmental samples, and further, from its original isolation in India during 1993–1994. This was accurately identified and segregated from other clinical aeromonads (A. hydrophila subsp. hydrophila, A. caviae, A. veronii biovars veronii and sobria, A. trota, A. schubertii and A. jandaei) by using biochemical key tests. The API 20 E profile for all strains included in A. hydrophila subsp. dhakensis was 7047125. The prevalence of this species in Spanish sources was higher for water (9.4%) than for feces (6%) or eels (1.3%). Isolates recovered as pure cultures from diseased eels were moderately virulent (LD50 of 3.3×106 CFU fish−1) to challenged eels in experimental trials. They were all resistant to ticarcillin, amoxicillin-clavuranic acid, cefoxitin, and imipenem, regardless of its source. Our data point to A. hydrophila subsp. dhakensis as an emerging pathogen for humans and fish in temperate countries. PMID:22472298

  9. Aeromonas hydrophila-associated septicemia in captive crocodiles (Crocodylus johnstoni and Crocodylus porosus).

    PubMed

    Roh, Yoon-Seok; Park, Heejin; Cho, Hyun-Ung; Cho, Ara; Islam, Mohammad Rafiqul; Cho, Ho-Seong; Lim, Chae Woong; Kim, Bumseok

    2011-12-01

    Five 25-yr-old crocodiles (Crocodylus johnstoni and Crocodylus porosus) were diagnosed with Aeromonas hydrophila-associated septicemia accelerated by improper thermoregulation. At necropsy, pulmonary congestion and pleural effusion were the main lesions in the thorax. Necrotizing enteritis, intestinal hemorrhage, fibrinous serositis, hepatitis, and pancreatitis were observed in the abdominal cavities of all five crocodiles. Aeromonas hydrophila was identified in the pleural effusions and abdominal ascites of all necropsied crocodiles by using an API system 20NE. Aeromonas hydrophila infection and evaluation of virulence were confirmed by polymerase chain reaction targeting the 16S rRNA and extracellular hemolysin gene. The crocodiles in the present case were housed in an indoor facility at a private zoo that failed to optimize land and water portions of the enclosure, exposing the animals to impeded thermoregulation, and it is suggested that the pathogenesis was accelerated by the improper thermoregulation-induced stress. This is the first description of A. hydrophila pathogenicity associated with impeded thermoregulation in reptiles.

  10. Inactivation of Aeromonas hydrophila by Fe(II)-related-radical generation in oxidizing groundwaters.

    PubMed Central

    Kersters, I; Verstraete, W

    1996-01-01

    The survival of Aeromonas hydrophila AWWX1 in filter-sterilized phreatic groundwaters was studied by using viable counts. Aeromonas counts rapidly decreased 2 to 3 log units in oxidizing raw groundwaters from Snellegem and Beernem, Belgium (Snellegem-raw and Beernem-raw, respectively), containing high concentrations of Fe2+ (460 to 1,070 microM). The rapid decline in viable counts of Aeromonas cells in the oxidizing raw groundwater of Snellegem was prevented by the addition of an Fe2+ chelator (2,2'-dipyridyl) or compounds (i.e., ascorbic acid and catalase) that act on toxic oxygen species. The results suggest that free radicals, generated spontaneously in oxidizing Fe2+-containing groundwaters, caused the inactivation of A. hydrophila AWWX1. Evidence that free radicals are generated under the given conditions was provided by the observation that propylphosphonic acid, a compound which is very susceptible to radicals, was degraded upon addition to these waters. A. hydrophila PWBS, Pseudomonas fluorescens P17, Spirillum strain NOX, and heterotrophs showed decreases in culturability in filter-sterilized Snellegem-raw water similar to that shown by A. hydrophila AWWX1. These findings indicate that free radicals generated in Fe2+-containing groundwaters upon aeration are capable of inactivating various bacterial species. PMID:8795217

  11. Recombinant goose-type lysozyme in channel catfish: lysozyme activity and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection.

    PubMed

    Pridgeon, Julia W; Klesius, Phillip H; Dominowski, Paul J; Yancey, Robert J; Kievit, Michele S

    2013-10-01

    The objectives of this study were: 1) to investigate whether recombinant channel catfish lysozyme-g (CC-Lys-g) produced in Escherichia coli expression system possesses any lysozyme activity; and 2) to evaluate whether channel catfish lysozyme-g plasmid DNA could be used as an immunostimulant to protect channel catfish against Aeromonas hydrophila infection. Recombinant CC-Lys-g produced in E. coli expression system exhibited significant (P < 0.05) lytic activity against Gram-positive Micrococcus lysodeikticus and Gram-negative A. hydrophila. When pcDNA3.2-vectored recombinant channel catfish lysozyme-g (pcDNA-Lys-g) was transfected in channel catfish gill cells G1B, the over-expression of pcDNA-Lys-g offered significant (P < 0.05) protection to G1B cells against A. hydrophila infection. When channel catfish were intraperitoneally injected with pcDNA-Lys-g along with an adjuvant QCDCR, the transcriptional level of Lys-g was significantly (P < 0.05) increased. When pcDNA-Lys-g injected fish was challenged with a highly virulent A. hydrophila strain AL-09-71, pcDNA-Lys-g offered 100% protection to channel catfish at two days post DNA injection. Macrophages of fish injected with pcDNA-Lys-g produced significantly (P < 0.05) higher amounts of reactive oxygen species and nitric oxide than that of fish injected with pcDNA vector alone at two days post DNA injection. Taken together, our results suggest that pcDNA-Lys-g could be used as a novel immunostimulant to offer immediate protection to channel catfish against A. hydrophila infection.

  12. Bacillus amyloliquefaciens G1: A Potential Antagonistic Bacterium against Eel-Pathogenic Aeromonas hydrophila

    PubMed Central

    Cao, Haipeng; He, Shan; Wei, Ruopeng; Diong, Marek; Lu, Liqun

    2011-01-01

    Recent studies have revealed that the use of probiotics is an alternative to control marine aeromonas. However, few probiotics are available against Aeromonas hydrophila infections in eels. In the present study, a potential antagonistic strain G1 against the eel-pathogenic A. hydrophila was isolated from sediment underlying brackish water. Its extracellular products with antibacterial activities were shown to be stable under wide range of pH, temperature, and proteinase K. It was initially identified as Bacillus amyloliquefaciens using API identification kits and confirmed to be B. amyloliquefaciens strain (GenBank accession number DQ422953) by phylogenetic analysis. In addition, it was shown to be safe for mammalians, had a wide anti-A. hydrophila spectrum, and exhibited significant effects on inhibiting the growth of the eel-pathogenic A. hydrophila both in vitro and in vivo. To the best of our knowledge, this is the first report on a promising antagonistic Bacillus amyloliquefaciens strain from brackish water sediment against eel-pathogenic A. hydrophila. PMID:21754944

  13. Evidence for bacterial chemotaxis to cyanobacteria from a radioassay technique. [Lyngbya birgei; Aphanizomenon flos-aquae; Aeromonas hydrophila

    SciTech Connect

    Kangatharalingam, N.; Wang, Lizhu; Priscu, J.C. )

    1991-08-01

    Lyngbya birgei and Aphanizomenon flos-aquae elicited a significant chemotactic attraction of Aeromonas hydrophila compared with controls lacking cyanobacteria. There was a positive exponential relationship between biomass (chlorophyll a) of L. birgei and A. flos-aquae and chemotactic attraction of A. hydrophila. The assay equipment was simple and reliable and could be used to study bacterial chemotaxis in other species in situ.

  14. Aeromonas hydrophila meningitis and fulminant sepsis in preterm newborn: A case report and review of literature.

    PubMed

    Kali, A; Kalaivani, R; Charles, Pmv; Seetha, K S

    2016-01-01

    Neonatal meningitis is a lethal infection occurring in the 1st month of life. The risk of developing permanent neurological sequels is high among the neonates who survive. Bacterial pathogens are commonly associated with this condition. Aeromonas is a Gram-negative bacteria of aquatic habitat. Although isolation of Aeromonas species from neonates with blood stream infection is infrequently reported, neonatal meningitis caused by Aeromonas is exceedingly rare. We present a case of fulminant sepsis and meningitis caused by Aeromonas hydrophila in a preterm newborn male. The bacteria was isolated in culture from blood and cerebrospinal fluid. In spite of targeted antibiotics and supportive therapy, the baby failed to respond and died on the 12th day of life.

  15. Dropped head syndrome in a patient with Aeromonas hydrophila-induced septic arthritis of the shoulder.

    PubMed

    Wang, Sheng-Huei; Juan, Chun-Jung; Lin, Te-Yu

    2014-12-01

    Septic arthritis of the shoulder is very rarely caused by Aeromonas hydrophila. We present the case of a 45-year-old woman who presented with symptoms of painful shoulder for 2 weeks and dropped head for 1 week prior to admission. A. hydrophila was isolated from a culture of purulent synovial fluid. Magnetic resonance imaging revealed profuse abscess collection between the right infraspinatus muscle and trapezius muscle and swelling of the right and left paraspinatus muscles, which suggested myositis-related dropped head syndrome. After surgery with arthrotomy, function of the shoulder and neck extensors was significantly improved.

  16. Effects of refrigeration and alcohol on the load of Aeromonas hydrophila in oysters.

    PubMed

    Birkenhauer, J M; Oliver, J D

    2002-03-01

    Members of the bacterial genus Aeromonas are widely distributed throughout the environment and are readily cultured from a variety of foods. One member of this genus, Aeromonas hydrophila, has been reputed to be a significant cause of gastrointestinal disease. In this study, we examined the effects of refrigeration and alcohol on the level of A. hydrophila in oysters. Specifically, vodka was examined because it is used by the food service industry in preparation of Oysters Romanoff. One set of oysters was shucked on receipt, whereas others were refrigerated intact for 7 days at 5 degrees C. The oysters were blended and the numbers of A. hydrophila present determined using starch ampicillin agar. Oysters were also shucked and placed on the half shell with 5 ml of vodka for 10 min. The oysters were then washed and presumptive A. hydrophila levels determined in both the washate and homogenate. On the day of purchase, the average number of presumptive A. hydrophila found was 7.6 x 10(4) CFU/g of oyster meat. After 7 days of refrigeration, the average number had increased to 3.2 x 10(5) CFU/g of oyster meat. In the oysters treated with vodka, the average number of A. hydrophila present internally was 9.9 x 10(4) with high numbers (10(3) to 10(4)) isolated from the oyster surface. From these data, it is clear that refrigeration and alcohol treatment are not sufficient to reduce loads of A. hydrophila in or on oysters.

  17. Evaluation of an in vitro cell assay to select attenuated bacterial mutants of Aeromonas hydrophila and Edwardsiella tarda to channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Both Aeromonas hydrophila (causative agent of motile aeromonas septicemia) and Edwardsiella tarda (causative agent of enteric septicemia) are Gram-negative bacteria widely distributed in aquatic environments, affecting many fish species worldwide, including channel catfish and tilapia. To control ba...

  18. Aeromonas Hydrophila Endocarditis with Ruptured Mycotic Aneurysm of Right Renal Artery

    PubMed Central

    Pugliese, Maria Elena; Falcone, Marco; Oliva, Alessandra; Faccenna, Federico; D’Aluisio, Denise; Morelli, Sergio

    2016-01-01

    Aeromonas hydrophila has been infrequently reported as a cause of infection in humans. It has been associated with a variety of clinical syndromes but Aeromonas-related endocarditis is extremely rare. We present the case of a 76-year-old diabetic patient who was admitted to our hospital due to severe lumbar pain resistant to nonsteroidal anti-inflammatory drugs accompanied by fever (up to 38.5°C). The vital signs were normal and the physical examination was unremarkable except for tenderness over right flank. Laboratory investigation showed a mild leukocytosis (white blood cell count of 11,360×106/L) with elevation of inflammatory markers. Cardiac ultrasound showed a large vegetation on the mitral valve. Abdominal computed tomogrpahy revealed a ruptured aneurysm of the right renal artery. Multiple sets of blood culture grew A. hydrophila.

  19. Preparation of Metabolically Active Staphylococcus aureus Protoplasts by Use of the Aeromonas hydrophila Lytic Enzyme

    PubMed Central

    Coles, N. W.; Gross, R.

    1973-01-01

    Stable, metabolically active protoplasts of Staphylococcus aureus have been prepared by the use of a staphylolytic enzyme produced by Aeromonas hydrophila. Respiratory and glycolytic rates and synthesis of nucleic acids, protein, and lipid in these protoplasts, stabilized variously in 1.1 M sucrose, 0.37 M sodium succinate, or 0.37 M sodium sulfate, have been shown to be comparable with the same parameters in intact cells under the same conditions. Images PMID:4728270

  20. Aeromonas hydrophila typing scheme based on patterns of agglutination with erythrocytes and yeast cells.

    PubMed Central

    Adams, D; Atkinson, H M; Woods, W H

    1983-01-01

    An agglutination typing scheme has been developed for strains of Aeromonas hydrophila. Primary agglutination typing is based on testing agar-grown A. hydrophila cells with human, horse, rat, and guinea pig erythrocytes and Saccharomyces cerevisiae cells. Further subdivision of primary groups is based firstly on whether yeast cell agglutination is inhibited by a D-mannose polymer, yeast mannan, and secondly on patterns of inhibition of hemagglutination by yeast mannan and the monomeric sugars L-fucose, D-galactose, and D-mannose. A total of 320 isolates were tested, and these were divisible into 39 distinct types on the basis of this scheme. Application of this typing scheme in the future to isolates of A. hydrophila known to be associated with human infection may enable correlations to be made between particular agglutination types and human pathogenicity. PMID:6841579

  1. Classification of a Hypervirulent Aeromonas hydrophila Pathotype Responsible for Epidemic Outbreaks in Warm-Water Fishes.

    PubMed

    Rasmussen-Ivey, Cody R; Hossain, Mohammad J; Odom, Sara E; Terhune, Jeffery S; Hemstreet, William G; Shoemaker, Craig A; Zhang, Dunhua; Xu, De-Hai; Griffin, Matt J; Liu, Yong-Jie; Figueras, Maria J; Santos, Scott R; Newton, Joseph C; Liles, Mark R

    2016-01-01

    Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the People's Republic of China and the United States (US). Multiple lines of evidence indicate US catfish and Asian carp isolates of A. hydrophila affiliated with sequence type 251 (ST251) share a recent common ancestor. To address the genomic context for the putative intercontinental transfer and subsequent geographic spread of this pathogen, we conducted a core genome phylogenetic analysis on 61 Aeromonas spp. genomes, of which 40 were affiliated with A. hydrophila, with 26 identified as epidemic strains. Phylogenetic analyses indicate all ST251 strains form a coherent lineage affiliated with A. hydrophila. Within this lineage, conserved genetic loci unique to A. hydrophila were identified, with some genes present in consistently higher copy numbers than in non-epidemic A. hydrophila isolates. In addition, results from analyses of representative ST251 isolates support the conclusion that multiple lineages are present within US vAh isolated from Mississippi, whereas vAh isolated from Alabama appear clonal. This is the first report of genomic heterogeneity within US vAh isolates, with some Mississippi isolates showing closer affiliation with the Asian grass carp isolate ZC1 than other vAh isolated in the US. To evaluate the biological significance of the identified heterogeneity, comparative disease challenges were conducted with representatives of different vAh genotypes. These studies revealed that isolate ZC1 yielded significantly lower mortality in channel catfish, relative to Alabama and Mississippi vAh isolates. Like other Asian vAh isolates, the ZC1 lineage contains all core genes for a complete type VI secretion system (T6SS). In contrast, more virulent US isolates

  2. Classification of a Hypervirulent Aeromonas hydrophila Pathotype Responsible for Epidemic Outbreaks in Warm-Water Fishes

    PubMed Central

    Rasmussen-Ivey, Cody R.; Hossain, Mohammad J.; Odom, Sara E.; Terhune, Jeffery S.; Hemstreet, William G.; Shoemaker, Craig A.; Zhang, Dunhua; Xu, De-Hai; Griffin, Matt J.; Liu, Yong-Jie; Figueras, Maria J.; Santos, Scott R.; Newton, Joseph C.; Liles, Mark R.

    2016-01-01

    Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the People's Republic of China and the United States (US). Multiple lines of evidence indicate US catfish and Asian carp isolates of A. hydrophila affiliated with sequence type 251 (ST251) share a recent common ancestor. To address the genomic context for the putative intercontinental transfer and subsequent geographic spread of this pathogen, we conducted a core genome phylogenetic analysis on 61 Aeromonas spp. genomes, of which 40 were affiliated with A. hydrophila, with 26 identified as epidemic strains. Phylogenetic analyses indicate all ST251 strains form a coherent lineage affiliated with A. hydrophila. Within this lineage, conserved genetic loci unique to A. hydrophila were identified, with some genes present in consistently higher copy numbers than in non-epidemic A. hydrophila isolates. In addition, results from analyses of representative ST251 isolates support the conclusion that multiple lineages are present within US vAh isolated from Mississippi, whereas vAh isolated from Alabama appear clonal. This is the first report of genomic heterogeneity within US vAh isolates, with some Mississippi isolates showing closer affiliation with the Asian grass carp isolate ZC1 than other vAh isolated in the US. To evaluate the biological significance of the identified heterogeneity, comparative disease challenges were conducted with representatives of different vAh genotypes. These studies revealed that isolate ZC1 yielded significantly lower mortality in channel catfish, relative to Alabama and Mississippi vAh isolates. Like other Asian vAh isolates, the ZC1 lineage contains all core genes for a complete type VI secretion system (T6SS). In contrast, more virulent US isolates

  3. The LBP gene and its association with resistance to Aeromonas hydrophila in tilapia.

    PubMed

    Fu, Gui Hong; Liu, Feng; Xia, Jun Hong; Yue, Gen Hua

    2014-12-01

    Resistance to pathogens is important for the sustainability and profitability of food fish production. In immune-related genes, the lipopolysaccharide-binding protein (LBP) gene is an important mediator of the inflammatory reaction. We analyzed the cDNA and genomic structure of the LBP gene in tilapia. The full-length cDNA (1901 bp) of the gene contained a 1416 bp open reading frame, encoding 471 amino acid residues. Its genomic sequence was 5577 bp, comprising 15 exons and 14 introns. Under normal conditions, the gene was constitutively expressed in all examined tissues. The highest expression was detected in intestine and kidney. We examined the responses of the gene to challenges with two bacterial pathogens Streptcoccus agalactiae and Aeromonas hydrophila. The gene was significantly upregulated in kidney and spleen post-infection with S. agalactiae and A. hydrophila, respectively. However, the expression profiles of the gene after the challenge with the two pathogens were different. Furthermore, we identified three SNPs in the gene. There were significant associations (p < 0.05) of two of the three SNPs with the resistance to A. hydrophila, but not with the resistance to S. agalactiae or growth performance. These results suggest that the LBP gene is involved in the acute-phase immunologic response to the bacterial infections, and the responses to the two bacterial pathogens are different. The two SNPs associated with the resistance to A. hydrophila may be useful in the selection of tilapia resistant to A. hydrophila.

  4. Novel insights into the pathogenicity of epidemic Aeromonas hydrophila ST251 clones from comparative genomics

    PubMed Central

    Pang, Maoda; Jiang, Jingwei; Xie, Xing; Wu, Yafeng; Dong, Yuhao; Kwok, Amy H. Y.; Zhang, Wei; Yao, Huochun; Lu, Chengping; Leung, Frederick C.; Liu, Yongjie

    2015-01-01

    Outbreaks in fish of motile Aeromonad septicemia (MAS) caused by Aeromonas hydrophila have caused a great concern worldwide. Here, for the first time, we provide two complete genomes of epidemic A. hydrophila strains isolated in China. To gain an insight into the pathogenicity of epidemic A. hydrophila, we performed comparative genomic analyses of five epidemic strains belonging to sequence type (ST) 251, together with the environmental strain ATCC 7966T. We found that the known virulence factors, including a type III secretion system, a type VI secretion system and lateral flagella, are not required for the high virulence of the ST251 clonal group. Additionally, our work identifies three utilization pathways for myo-inositol, sialic acid and L-fucose providing clues regarding the factors that underlie the epidemic and virulent nature of ST251 A. hydrophila. Based on the geographical distribution and biological resources of the ST251 clonal group, we conclude that ST251 is a high-risk clonal group of A. hydrophila which may be responsible for the MAS outbreaks in China and the southeastern United States. PMID:26014286

  5. Establishment of Infection Models in Zebrafish Larvae (Danio rerio) to Study the Pathogenesis of Aeromonas hydrophila

    PubMed Central

    Saraceni, Paolo R.; Romero, Alejandro; Figueras, Antonio; Novoa, Beatriz

    2016-01-01

    Aeromonas hydrophila is a Gram-negative opportunistic pathogen of fish and terrestrial animals. In humans, A. hydrophila mainly causes gastroenteritis, septicaemia, and tissue infections. The mechanisms of infection, the main virulence factors and the host immune response triggered by A. hydrophila have been studied in detail using murine models and adult fish. However, the great limitation of studying adult animals is that the animal must be sacrificed and its tissues/organs extracted, which prevents the study of the infectious processes in the whole living animal. Zebrafish larvae are being used for the analysis of several infectious diseases, but their use for studying the pathogenesis of A. hydrophila has never been explored. The great advantage of zebrafish larvae is their transparency during the first week after fertilization, which allows detailed descriptions of the infectious processes using in vivo imaging techniques such as differential interferential contrast (DIC) and fluorescence microscopy. Moreover, the availability of fluorescent pathogens and transgenic reporter zebrafish lines expressing fluorescent immune cells, immune marker genes or cytokines/chemokines allows the host–pathogen interactions to be characterized. The present study explores the suitability of zebrafish larvae to study the pathogenesis of A. hydrophila and the interaction mechanisms between the bacterium and the innate immune responses through an infection model using different routes for infection. We used an early-embryo infection model at 3 days post-fertilization (dpf) through the microinjection of A. hydrophila into the duct of Cuvier, caudal vein, notochord, or muscle and two bath infection models using 4 dpf healthy and injured larvae. The latter resembled the natural conditions under which A. hydrophila produces infectious diseases in animals. We compared the cellular processes after infection in each anatomical site by confocal fluorescence imaging and determined the

  6. Hemolytic and proteolytic activities of Aeromonas hydrophila and Aeromonas veronii biovar sobria in broth and salmon extract at different temperatures.

    PubMed

    González-Rodríguez, María-Nieves; Santos, Jesús A; Otero, Andrés; García-López, Maria-Luisa

    2004-02-01

    Expression of hemolytic and proteolytic activities throughout the growth cycle was investigated with two enterotoxic aeromonad strains assigned to the species Aeromonas hydrophila and Aeromonas veronii biovar sobria. Although growth kinetic data were dependent on strain, temperature, and substrate, maximum populations attained were higher than 9 log CFU/ml in aerated tryptone soya broth plus yeast extract (TSBYE) and salmon extract within the range 4 to 28 degrees C. For both strains in TSBYE, variable amounts of hemolytic activity were first detected at any temperature when aeromonad counts were over 9 log CFU/ml. Afterwards, this activity increased up to similar levels (109 to 112 hemolytic units per ml) without a significant increase in populations. Salmon extract supported hemolysin synthesis at 28 but not 4 degrees C. Proteolytic activity of the A. hydrophila strain was only expressed in salmon extract at 28 degrees C, whereas A. veronii biovar sobria did at 28 degrees C in both substrates and at 10 degrees C in TSBYE.

  7. Clinical and biochemical significance of toxin production by Aeromonas hydrophila.

    PubMed

    Kindschuh, M; Pickering, L K; Cleary, T G; Ruiz-Palacios, G

    1987-05-01

    Production of cytotoxin and enterotoxin by Aeromonas strains obtained from stools of 50 children in Mexico and Texas and from blood of 9 children with sepsis was determined. Results were correlated with clinical features of infected children as well as with biochemical traits of Aeromonas strains. Cytotoxin was produced by 40 of 42 Aeromonas strains (95%) isolated from stools of children with diarrhea, by all 8 isolates from stools of well children, and by all 9 isolates from children with sepsis. There was no difference in the quantities (amount of cytotoxin per milligram of protein required to kill 50% of the cells) of cytotoxin produced and in clinical manifestations among the groups. None of the isolates produced a toxin that could be neutralized by antiserum raised against Shiga toxin produced by Shigella dysenteriae 1 60R. Heat-labile-like enterotoxin (LT) was produced by 26 of 42 stool isolates (62%), while only 1 of the 42 isolates (2%) produced enterotoxinlike activity in suckling mice; 65% of the cytotoxin-producing strains also produced an LT-like material. All strains from blood produced LT-like material, and 2 of 6 (33%) produced activity in suckling mice. All strains produced hemolysin; 37 of 57 (65%) were Voges-Proskauer positive; 27 of 57 (47%) were lysine decarboxylase positive by API 20E strips, none were positive for lysine decarboxylose production by lysin-iron agar slants at 24 h, but 17 of 54 (31%) were positive at 48 h. There was no correlation between biochemical reactions and enterotoxin or cytotoxin production. There appears to be no correlation between toxin production by Aeromonas spp. and gastroenteritis.

  8. Orally administered thermostable N-acyl homoserine lactonase from Bacillus sp. strain AI96 attenuates Aeromonas hydrophila infection in zebrafish.

    PubMed

    Cao, Yanan; He, Suxu; Zhou, Zhigang; Zhang, Meichao; Mao, Wei; Zhang, Huitu; Yao, Bin

    2012-03-01

    N-Acylated homoserine lactone (AHL) lactonases are capable of degrading signal molecules involved in bacterial quorum sensing and therefore represent a new approach to control bacterial infection. Here a gene responsible for the AHL lactonase activity of Bacillus sp. strain AI96, 753 bp in length, was cloned and then expressed in Escherichia coli. The deduced amino acid sequence of Bacillus sp. AI96 AiiA (AiiA(AI96)) is most similar to those of other Bacillus sp. AHL lactonases (~80% sequence identity) and was consequently categorized as a member of the metallo-β-lactamase superfamily. AiiA(AI96) maintains ~100% of its activity at 10°C to 40°C at pH 8.0, and it is very stable at 70°C at pH 8.0 for at least 1 h; no other Bacillus AHL lactonase has been found to be stable under these conditions. AiiA(AI96) resists digestion by proteases and carp intestinal juice, and it has broad-spectrum substrate specificity. The supplementation of AiiA(AI96) into fish feed by oral administration significantly attenuated Aeromonas hydrophila infection in zebrafish. This is the first report of the oral administration of an AHL lactonase for the efficient control of A. hydrophila.

  9. Outbreak of Abortions and Infertility in Thoroughbred Mares Associated with Waterborne Aeromonas hydrophila.

    PubMed

    Singh, B R; Gulati, B R; Virmani, Nitin; Chauhan, Mamta

    2011-06-01

    At a thoroughbred equine breeding farm near Hissar (Haryana), three mares aborted in their seventh month of pregnancy. The vaginal swabs of all aborted mares, and stomach contents, heart blood, liver, spleen and placenta of aborted fetuses yielded pure culture of Aeromonas hydrophila. In addition, A. hydrophila was also isolated from the vaginal swabs of three repeat breeding mares and faecal sample of a diarrheic foal. The source of infection was possibly water supply as all the water samples collected from taps, mother tank and storage tank were found to be positive for A. hydrophila. The antibiogram of all the isolates was similar showing resistance to ampicillin, carbenicillin, gentamicin, kanamycin and amikacin but sensitive to chloramphenicol, ciprofloxacin, cefuroxime, ceftriaxone, cotrimoxazole, cotrimazine, nitrofurantoin, streptomycin and tetracycline. All the 20 sera samples collected from three aborted and three repeat breeding, and eight in-contact mares, a diarrheic foal, three cows and two male buffaloes maintained at the same farm contained antibodies against A. hydrophila with titres ranging from 80 to 640. The water supply was instantly chlorinated using 0.05% sodium hypochlorite for three consecutive days and all the culturally positive mares were treated with intravaginal administration of 1 g ciprofloxacin, while the foal was given nitrofurantoin for three days. After one month, A. hydrophila could not be isolated either from mares or from their environment and antibody titre in all the seropositive animals showed a declining trend. Later, all the aborted and repeat breeding mares were confirmed to be pregnant. Thus, the present study indicated that water-borne A. hydrophila might be associated with equine abortions and infertility, and diarrhea in newborn foals.

  10. Proteomic Analysis Reveals That Metabolic Flows Affect the Susceptibility of Aeromonas hydrophila to Antibiotics

    PubMed Central

    Yao, Zujie; Li, Wanxin; Lin, Yi; Wu, Qian; Yu, Feifei; Lin, Wenxiong; Lin, Xiangmin

    2016-01-01

    The overuse of antibiotics results in the development of antibiotic resistance and limits the useful life of these drugs in fighting bacteria, including Aeromonas hydrophila, a well-known opportunistic pathogen that causes serious infections in fish and other animals. In this study, we investigated the adaptive resistance mechanism in A. hydrophila by multiple proteomic methods. Dimethyl labeling and label-free methods were performed to compare the differential expression of proteins in response to various doses of oxytetracycline (OXY). The results point to the conclusions that, in response to OXY stress, translational processes increase the abundance of these proteins whereas largely central metabolic pathways decrease their abundance. To confirm our hypothesis, various exogenous metabolites were compounded with OXY, and the resulting survival capabilities were measured. Results show that 7 metabolites (malic acid, serine, methionine, etc.) significantly decreased the survival capabilities of A. hydrophila in the presence of OXY, whereas 4 metabolites (arginine, lysine, tyrosine, etc.) did the opposite. Further investigation suggests that a compound comprising exogenous metabolites in combination with various antibiotics could have a significant bactericidal effect and might come into widespread use, especially together with tetracycline antibiotics. These findings may provide new clues to the antimicrobial treatment of A. hydrophila infection. PMID:27991550

  11. An Asian Origin of Virulent Aeromonas hydrophila Responsible for Disease Epidemics in United States-Farmed Catfish

    PubMed Central

    Hossain, Mohammad J.; Sun, Dawei; McGarey, Donald J.; Wrenn, Shannon; Alexander, Laura M.; Martino, Maria Elena; Xing, Ye; Terhune, Jeffery S.

    2014-01-01

    ABSTRACT Since 2009, catfish farming in the southeastern United States has been severely impacted by a highly virulent and clonal population of Aeromonas hydrophila causing motile Aeromonas septicemia (MAS) in catfish. The possible origin of this newly emerged highly virulent A. hydrophila strain is unknown. In this study, we show using whole-genome sequencing and comparative genomics that A. hydrophila isolates from diseased grass carp in China and catfish in the United States have highly similar genomes. Our phylogenomic analyses suggest that U.S. catfish isolates emerged from A. hydrophila populations of Asian origin. Furthermore, we identified an A. hydrophila strain isolated in 2004 from a diseased catfish in Mississippi, prior to the onset of the major epidemic outbreaks in Alabama starting in 2009, with genomic characteristics that are intermediate between those of the Asian and Alabama fish isolates. Investigation of A. hydrophila strain virulence demonstrated that the isolate from the U.S. catfish epidemic is significantly more virulent to both channel catfish and grass carp than is the Chinese carp isolate. This study implicates the importation of fish or fishery products into the United States as the source of highly virulent A. hydrophila that has caused severe epidemic outbreaks in United States-farmed catfish and further demonstrates the potential for invasive animal species to disseminate bacterial pathogens worldwide. PMID:24895303

  12. Molecular Characterization of a Functional Type VI Secretion System from a Clinical Isolate of Aeromonas hydrophila

    PubMed Central

    Suarez, Giovanni; Sierra, Johanna C.; Sha, Jian; Wang, Shaofei; Erova, Tatiana E.; Fadl, Amin A.; Foltz, Sheri M.; Horneman, Amy J.; Chopra, Ashok K.

    2008-01-01

    Our laboratory recently molecularly characterized the type II secretion system (T2SS)- associated cytotoxic enterotoxin (Act) and the T3SS-secreted AexU effector from a diarrheal isolate SSU of Aeromonas hydrophila. The role of these toxin proteins in the pathogenesis of A. hydrophila infections was subsequently delineated in in vitro and in vivo models. In this study, we characterized the new type 6 secretion system (T6SS) from isolate SSU of A. hydrophila and demonstrated its role in bacterial virulence. Study of the role of T6SS in bacterial virulence is in its infancy, and there are, accordingly, only limited, recent reports directed toward a better understanding its role in bacterial pathogenesis. We have provided evidence that the virulence-associated secretion (vas) genes vasH (Sigma 54-dependent transcriptional regulator) and vasK (encoding protein of unknown function) are essential for expression of the genes encoding the T6SS and/or they constituted important components of the T6SS. Deletion of the vasH gene prevented expression of the potential translocon hemolysin coregulated protein (Hcp) encoding gene from bacteria, while the vasK gene deletion prevented secretion but not translocation of Hcp into host cells. The secretion of Hcp was independent of the T3SS and the flagellar system. We demonstrated that secreted Hcp could bind to the murine RAW 264.7 macrophages from outside, in addition to its ability to be translocated into host cells. Further, the vasH and vasK mutants were less toxic to murine macrophages and human epithelial HeLa cells, and these mutants were more efficiently phagocytosed by macrophages. We also provided evidence that the expression of the hcp gene in the HeLa cell resulted in apoptosis of the host cells. Finally, the vasH and vasK mutants of A. hydrophila were less virulent in a septicemic mouse model of infection, and animals immunized with recombinant Hcp were protected from subsequent challenge with the wild-type (WT

  13. Antibacterial Activity of Various Plants Extracts Against Antibiotic-resistant Aeromonas hydrophila

    PubMed Central

    Al Laham, Shaza Anwar; Al Fadel, Frdoos Mohammad

    2014-01-01

    Background: Aeromonas hydrophila cause one of the most important diseases in fishes and lead to economic losses, and may be contaminated human beings. Objectives: The current research aimed to investigate the anti-bacterial activity shown by the extracts prepared from different parts of Olea europea, Myrtus communis, Thymus vulgaris, Rosmarinuis officinalis, and Achillea falcata that grow in Syria against A. hydrophila that causes the most dangerous bacterial diseases in fish. Materials and Methods: The study was performed in four stages: First of all, the presence of A. hydrophila was investigated in 450 Samples of Cyprinus Carpio fish using blood agar, Trypticase soya agar, and Analytical Profile Index (API20E). Secondly, the plants extract was obtained using water, absolute alcohol, then ether using Soxhlet extraction apparatus and rotary vacuum evaporator. Thirdly, the antibacterial activity of some antibiotics on these bacteria was evaluated by disk diffusion method. Finally, the antibacterial effect of the extracts was determined by disk diffusion method. Results: The studied antibiotics showed no antibacterial activity against these bacteria, except amikacin which had an acceptable effectiveness. However, the ethanol extracts of the studied plants revealed different antibacterial effects against A. hydrophila which showed antibiotic resistant. T. vulgaris extract had the strongest effect, whereas O. europea extract had the weakest activity. The water and ether petroleum extracts had no antibacterial activities. Conclusions: Ethanol extracts of the studied plants had different antibacterial effects against antibiotic-resistant A. hydrophila. T. vulgaris had the highest activity, R. officinalis had the second, and M. communis and A. falcate were in the third place, while the O. europea had the weakest antibacterial activity. PMID:25368797

  14. Involvement of peroxinectin in the defence of red swamp crayfish Procambarus clarkii against pathogenic Aeromonas hydrophila.

    PubMed

    Dong, Chaohua; Wei, Zhu; Yang, Guanpin

    2011-06-01

    Cell adhesion factors are important immune components for invertebrate to immobilize, phagocytose or encapsulate invasive microorganisms and foreign particles. In this study, a new cell adhesion factor, peroxinectin (refered as Pcpxin) was isolated from hemocytes of red swamp crayfish (Procambarus clarkii). The full-length cDNA of Pcpxin was 3014 bp encoding a protein of 819 amino acid residues with a predicted molecular weight of 89.0 kDa and a calculational isoelectric point of 6.93. The putative amino acid sequence contained a peroxidase domain and a signal peptide of 21 amino acid residues, and exhibited high identity to peroxinectin from Pacifastacus leniusculus (85%), Fenneropenaeus chinensis (62%) and Scylla serrata (58%), as well as peroxidase from Camponotus floridanus (40%), Pediculus humanus corporis (39%), and Culex quinquefasciatus (38%). Quantitative real time PCR revealed that mRNA expression of Pcpxin in hemocytes could be inhibited by challenge with heat-killed Aeromonas hydrophila, suggesting that Pcpxin was involved in immune responses to A. hydrophila. RNA interference (RNAi) experiment demonstrated that silencing Pcpxin significantly reduced the survival rate of red swamp crayfishes after challenge with A. hydrophila, which indicated that Pcpxin was important for P. clarkii to survive A. hydrophila infection. Moreover, silencing Pcpxin inhibited the up-regulation of crustin1 and lysozyme expression in response to challenge with heat-killed A. hydrophila. This result suggested that Pcpxin might participate in antibacterial peptide gene expression and thereby might be involved in signal transduction pathway regulating the expression of antibacterial peptide gene.

  15. Bioremediation and Detoxification of Synthetic Wastewater Containing Triarylmethane Dyes by Aeromonas hydrophila Isolated from Industrial Effluent

    PubMed Central

    Ogugbue, Chimezie Jason; Sawidis, Thomas

    2011-01-01

    Economical and bio-friendly approaches are needed to remediate dye-contaminated wastewater from various industries. In this study, a novel bacterial strain capable of decolorizing triarylmethane dyes was isolated from a textile wastewater treatment plant in Greece. The bacterial isolate was identified as Aeromonas hydrophila and was shown to decolorize three triarylmethane dyes tested within 24 h with color removal in the range of 72% to 96%. Decolorization efficiency of the bacterium was a function of operational parameters (aeration, dye concentration, temperature, and pH) and the optimal operational conditions obtained for decolorization of the dyes were: pH 7-8, 35°C and culture agitation. Effective color removal within 24 h was obtained at a maximum dye concentration of 50 mg/L. Dye decolorization was monitored using a scanning UV/visible spectrophotometer which indicated that decolorization was due to the degradation of dyes into non-colored intermediates. Phytotoxicity studies carried out using Triticum aestivum, Hordeum vulgare, and Lens esculenta revealed the triarylmethane dyes exerted toxic effects on plant growth parameters monitored. However, significant reduction in toxicity was obtained with the decolorized dye metabolites thus, indicating the detoxification of the dyes following degradation by Aeromonas hydrophila. PMID:21808740

  16. Chicken-type lysozyme in channel catfish: expression analysis, lysozyme activity, and efficacy as immunostimulant against Aeromonas hydrophila infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To understand whether chicken-type lysozyme (Lys-c) in channel catfish was induced by infection of Aeromonas hydrophila, the transcriptional levels of Lys-c in skin, gut, liver, spleen, posterior kidney, and blood cells in healthy channel catfish was compared to that in channel catfish infected with...

  17. Draft Genome Sequence of Pseudomonas mosselii Gil3, Isolated from Catfish and Antagonistic against Hypervirulent Aeromonas hydrophila

    PubMed Central

    Xu, De-Hai; Qiu, Junqiang; Rasmussen-Ivey, Cody R.; Liles, Mark R.; Beck, Benjamin H.

    2016-01-01

    Pseudomonas mosselii Gil3 was isolated from a catfish that survived from lethal challenge with hypervirulent Aeromonas hydrophila (vAh). When assayed in vitro, the bacterium showed antagonism against vAh. Sequence analysis revealed that the genome of P. mosselii Gil3 encodes numerous aromatic metabolism pathways and proteins for biosynthesis of antimicrobial compounds. PMID:27856595

  18. Draft genome sequence of Pseudomonas mosselii Gil3, isolated from catfish and antagonistic against hypervirulent Aeromonas hydrophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pseudomonas mosselii Gil3 was isolated from a catfish that survived from lethal challenge with hypervirulent Aeromonas hydrophila (vAh). When assayed in vitro, the bacterium showed antagonism against vAh. Sequence analysis revealed that the genome of P. mosselii Gil3 encodes numerous aromatic metabo...

  19. Chicken-type lysozyme in channel catfish: Expression analysis, lysozyme activity and efficacy as immunostimulant against Aeromonas hydrophila infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To understand whether chicken-type lysozyme (Lys-c) in channel catfish was induced by infection of Aeromonas hydrophila, the transcriptional levels of Lys-c in skin, gut, liver, spleen, posterior kidney, and blood cells in healthy channel catfish was compared to that in channel catfish infected with...

  20. Development and efficacy of novobiocin and rifampicin-resistant Aeromonas hydrophila as novel vaccines in channel catfish and Nile tilapia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three attenuated Aeromonas hydrophila vaccines were developed from the virulent 2009 West Alabama isolates through selection for resistance to both novobiocin and rifampicin. When channel catfish (Ictalurus punctatus) were IP injected with 4×105 colony-forming units (CFU) of the mutants, no fish die...

  1. An experimental vaccine against Aeromonas hydrophila can induce protection in rainbow trout, Oncorhynchus mykiss (Walbaum)

    USGS Publications Warehouse

    LaPatra, S.E.; Plant, K.P.; Alcorn, S.; Ostland, V.; Winton, J.

    2010-01-01

    A candidate vaccine against Aeromonas hydrophila in rainbow trout, Oncorhynchus mykiss, was developed using a bacterial lysate. To test the strength of protection, A. hydrophila challenge models were compared using injection into both the intraperitoneal (IP) cavity and the dorsal sinus (DS) with selected doses of live bacteria washed in saline or left untreated. Unlike the IP route, injection into the DS with either saline washed or unwashed cells resulted in consistent cumulative mortality and a dose response that could be used to establish a standard challenge having an LD50 of approximately 3 × 107 colony forming units per fish. Survivors of the challenge suffered significantly lower mortality upon re-challenge than naïve fish, suggesting a high level of acquired resistance was elicited by infection. Passive immunization using serum from hyper-immunized fish also resulted in significantly reduced mortality indicating protection can be transferred and that some portion of resistance may be antibody mediated. Vaccination of groups of rainbow trout with A. hydrophila lysate resulted in significant protection against a high challenge dose but only when injected along with Freund’s complete adjuvant. At a low challenge dose, mortality in all groups was low, but the bacterial lysate alone appeared to offer some protection.

  2. Quorum quenching bacteria Bacillus sp. QSI-1 protect zebrafish (Danio rerio) from Aeromonas hydrophila infection

    PubMed Central

    Chu, Weihua; Zhou, Shuxin; Zhu, Wei; Zhuang, Xiyi

    2014-01-01

    Quorum Sensing (QS) is a bacterial regulatory mechanism, which is responsible for controlling the expression of various biological macromolecules such as the virulence factors in a cell density-dependent manner. Disruption of the QS system of pathogens has been proposed as a new anti-infective strategy. Biodegradation of AHLs proves to be an efficient way to interrupt QS, since AHLs are the main family of QS autoinducers used in Gram negative bacteria. In this study, the effect of Bacillus sp. QSI-1 as an efficient quorum quencher on virulence factors production and biofilm formation of fish pathogen Aeromonas hydrophila was investigated. QSI-1 reduced the accumulation of AHLs but did not affect the growth of A. hydrophila YJ-1 when cocultured. In the result, the supernatant of QSI-1 showed significant inhibition of protease production (83.9%), hemolytic activity (77.6%) and biofilm formation (77.3%) in YJ-1. In biocontrol experiment, QSI-1 significantly reduced the pathogenicity of A. hydrophila strain YJ-1 in zebrafish (Danio rerio). The fish fed with QSI-1 was observed to have a relative percentage survival of 80.8%. Our results indicate that AHLs degrading bacteria should be considered as an alternative for antibiotics in aquaculture for the biocontrol of bacterial fish diseases. PMID:24962441

  3. Influence of glucose concentrations on biofilm formation, motility, exoprotease production, and quorum sensing in Aeromonas hydrophila.

    PubMed

    Jahid, Iqbal Kabir; Lee, Na-Young; Kim, Anna; Ha, Sang-Do

    2013-02-01

    Aeromonas hydrophila recently has received increased attention because it is opportunistic and a primary human pathogen. A. hydrophila biofilm formation and its control are a major concern for food safety because biofilms are related to virulence. Therefore, we investigated biofilm formation, motility inhibition, quorum sensing, and exoprotease production of this opportunistic pathogen in response to various glucose concentrations from 0.05 to 2.5% (wt/vol). More than 0.05% glucose significantly impaired (P < 0.05) quorum sensing, biofilm formation, protease production, and swarming and swimming motility, whereas bacteria treated with 0.05% glucose had activity similar to that of the control (0% glucose). A stage shift biofilm assay revealed that the addition of glucose (2.5%) inhibited initial biofilm formation but not later stages. However, addition of quorum sensing molecules N-3-butanoyl-DL-homoserine lactone and N-3-hexanoyl homoserine lactone partially restored protease production, indicating that quorum sensing is controlled by glucose concentrations. Thus, glucose present in food or added as a preservative could regulate acyl-homoserine lactone quorum sensing molecules, which mediate biofilm formation and virulence in A. hydrophila.

  4. Global protein expression profile response of planktonic Aeromonas hydrophila exposed to chlortetracycline.

    PubMed

    Li, Wanxin; Yao, Zujie; Zhang, Xiangyu; Huang, Fang; Lin, Wenxiong; Lin, Xiangmin

    2017-04-01

    The antibiotics resistance phenomena of Aeromonas hydrophila has become serious economic and public health problems for the world aquaculture industry and human health care. In this study, to investigate the instinct antibiotics adaptive mechanism of this pathogen, iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) based quantitative proteomics technologies were performed to compare the differential expression of A. hydrophila in planktonic status in response to chlortetracycline (CTC) stress and then identified total 1552 proteins including 285 altered proteins with 90 increasing and 195 decreasing abundance proteins. The following bioinformatics analysis showed that many metabolic metabolism pathways such as carbon metabolism, pyruvate metabolism, and glycolysis/gluconeogenesis were trend to down-regulated whereas β-Lactam resistance, RNA degradation, and amino acids biosynthesis processes were more likely to increase in CTC stress. The related pyruvate metabolism and β-Lactam resistance processes in mRNA level were further measured using the q-PCR method. Thus, an understanding of the behaviors of A. hydrophila in response to CTC would be helpful to reveal the antibiotics adaptive mechanism and for the development of novel antibiotics therapy.

  5. Non-specific immune response of bullfrog Rana catesbeiana to intraperitoneal injection of bacterium Aeromonas hydrophila

    NASA Astrophysics Data System (ADS)

    Zhang, Junjie; Zou, Wenzheng; Yan, Qingpi

    2008-08-01

    Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution (0.85%, w/v). Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection (dpi) for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase (SOD) activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection.

  6. Transcriptome signatures in common carp spleen in response to Aeromonas hydrophila infection.

    PubMed

    Jiang, Yanliang; Feng, Shuaisheng; Zhang, Songhao; Liu, Hong; Feng, Jianxin; Mu, Xidong; Sun, Xiaowen; Xu, Peng

    2016-10-01

    The common carp is an important aquaculture species that is worldwide distributed. Nowadays, intensive rearing in aquaculture increases the susceptibility of fish to various pathogens such as Aeromonas hydrophila, which has caused severe damage to carp production. However, systematic analysis on the host response of common carp against A. hydrophila is less studied. In order to better understand the common carp immune response process against bacteria at the global gene expression level, we examined transcriptional profiles of the common carp spleen at three timepoints following experimental infection with A. hydrophila. A total of 545 million 125-bp paired end reads were generated, and all trimmed clean reads were mapped onto the common carp whole genome sequence. Comparison of the transcriptomes between the treatment and control group fish revealed 2900 unigenes with significantly differential expression, including 732, 936, 928 genes up-regulated, and 248, 475, 700 genes down-regulated at 4 h, 12 h, 24 h post infection respectively. The captured significantly differentially expressed genes are mainly involved in the pathways including junction/adhesion, pathogen recognition, cell surface receptor signaling, and immune system process/defense response. Our study will provide fundamental information on molecular mechanism underlying the immune response of teleost against bacterial infection and might suggest strategies for selection of resistant strains of common carp in aquaculture.

  7. Aeromonas hydrophila induced head kidney macrophage apoptosis in Clarias batrachus involves the activation of calpain and is caspase-3 mediated.

    PubMed

    Banerjee, Chaitali; Goswami, Ramansu; Verma, Gaurav; Datta, Malabika; Mazumder, Shibnath

    2012-07-01

    The mechanism of macrophage cytotoxicity induced by Aeromonas hydrophila is yet unresolved. We observed A. hydrophila induces Head Kidney Macrophage (HKM) apoptosis in Clarias batrachus, as evident from Hoechst 33342 and AnnexinV-Propidium Iodide staining and presence of oligonucleosomal DNA ladder. Initiation of apoptosis required the bacteria to be alive, be actively phagocytosed into HKM and was dependent on host proteins. Elevated cytosolic calcium and consequent calpain activity that declined following pre-incubation with EGTA, verapamil and nifedipine implicates the role of calcium influx through voltage gated calcium channels and calpain in A. hydrophila-induced HKM apoptosis. Though, calpain-1 and -2 were involved, calpain-2 appeared to be more important in the process. EGTA, verapamil, nifedipine and calpain-2 inhibitor reduced caspase-3 activity and apoptosis. We conclude that A. hydrophila alters cytosolic calcium homeostasis initiating the activation of calpains, more specifically calpain-2, which leads to caspase-3 mediated HKM apoptosis in C. batrachus.

  8. Comparative study of semi-specific Aeromonas hydrophila and universal Pseudomonas fluorescens biosensors for BOD measurements in meat industry wastewaters.

    PubMed

    Raud, Merlin; Tenno, Toomas; Jõgi, Eerik; Kikas, Timo

    2012-04-05

    Aeromonas hydrophila P69.1 (A. hydrophila) was used to construct a semi-specific biosensor to estimate biochemical oxygen demand (BOD) in high fat and grease content wastewaters. A. hydrophila cells were grown in fat containing medium to induce necessary enzymes for transport and degradation of fatty substances. Universal biosensor based on non-specific Pseudomonas fluorescens P75 (P. fluorescens) was used to conduct comparison experiments. Biosensors were calibrated using OECD synthetic wastewater and steady-state method, subsequently several experiments with synthetic and industrial wastewaters were conducted. A linear range up to 45 mg l(-1) BOD(7) was gained using A. hydrophila biosensor, in comparison to 40 mg l(-1) BOD(7) obtained using P. fluorescens biosensors. The lower limit of detection was 5 mg l(-1) BOD(7). Service life of A. hydrophila and P. fluorescens biosensors were 110 and 115 days, respectively. The response time of the biosensors depended on the BOD(7) of measuring solution and was up to 20 min when analyzing different wastewaters. Both biosensors underestimated BOD in meat industry wastewater from 43% up to 71%, but more accurate results could be obtained with A. hydrophila biosensor. Semi-specific A. hydrophila biosensor was able to measure proportion of fat found in wastewater sample, while other refractory compounds remained undetectable to both biosensors.

  9. Effect of electron irradiation and packaging atmosphere on the survival of aeromonas hydrophila in minced poultry meat

    NASA Astrophysics Data System (ADS)

    Stecchini, M. L.; Sarais, I.; Del Torre, M.; Fuochi, P. G.

    1995-02-01

    Resistance to electron irradiation of Aeromonas hydrophila inoculated in minced poultry meat packed in presence of air or under vacuum was examined. Surviving bacteria were counted on starch ampicillin agar containing 100 μg/ml of ampicillin. Radiation resistance, expressed as D 10 values, was calculated from the survival curves and found to be 0.12 and 0.12 kGy in poultry meat packed in air or under vacuum respectively. Storage at 2°C of meat samples irradiated at 0.5 kGy further reduced the number of A. hydrophila.

  10. Protective immunity of grass carp immunized with DNA vaccine against Aeromonas hydrophila by using carbon nanotubes as a carrier molecule.

    PubMed

    Liu, Lei; Gong, Yu-Xin; Liu, Guang-Lu; Zhu, Bin; Wang, Gao-Xue

    2016-08-01

    To reduce the economic losses caused by diseases in aquaculture industry, more efficient and economic prophylactic measures should be urgently investigated. In this research, the effects of a novel functionalized single-walled carbon nanotubes (SWCNTs) applied as a delivery vehicle for DNA vaccine administration in juvenile grass carp against Aeromonas hydrophila were studied. Our results showed that SWCNTs loaded with DNA vaccine induced a better protection to juvenile grass carp against A. hydrophila. Moreover, SWCNTs conjugated with DNA vaccine provided significantly protective immunity compared with free DNA vaccine. Thereby, SWCNTs may be considered as a potential efficient DNA vaccine carrier to enhance the immunological activity.

  11. Aeromonas hydrophila and its relation with drinking water indicators of microbiological quality in Argentine.

    PubMed

    Fernández, M C; Giampaolo, B N; Ibañez, S B; Guagliardo, M V; Esnaola, M M; Conca, L; Valdivia, P; Stagnaro, S M; Chiale, C; Frade, H

    2000-01-01

    In Argentine, water municipal supplies disinfection is carried out by chlorine. We have isolated Aeromonas hydrophila from a chlorinated water supply in Buenos Aires that fulfilled Argentinean microbiological quality standards. It is an aquatic organism that could produce cytotoxins and enterotoxins associated with acute gastroenteritis and wound infections in human and hemorrhagic septicaemia of fish, reptiles and amphibians. The isolated strain produced protein bands at isoelectric point in the range of 4.6-5.3 lightly labile at 56 degrees C and heat labile at 100 degrees C non cholera antitoxin reactive in electrofocusing assay (IEF) and showed hemolytic activity thermolabile at 56 degrees C and 100 degrees C in 5% rabbit erythrocytes in phosphate buffered saline. A cytotoxic effect thermolabile at 56 degrees C and 100 degrees C and a cytotonic activity were demonstrated in Vero cell cultures. Survival assay of A. hydrophila and indicator organisms would show no correlation between their contamination sources. Biofilms production could explain the persistence of this gram negative pathogen organism in chlorinated tap water. A water system supply maintenance program and pasteurisation may be performed in spite of water chlorination.

  12. A 45-kDa acetylcholinesterase protoxin of Aeromonas hydrophila: purification and immunogenicity in fish.

    PubMed

    Pérez, M J; Rodríguez, L A; Fernández-Briera, A; Nieto, T P

    2002-05-21

    A rabbit antiserum to the 15-kDa acetylcholinesterase toxin neutralised the lethal effect of the 15-kDa toxin of Aeromonas hydrophila when injected into trout. However, immunisation of fish with the 15-kDa toxoid failed to induce an antibody response, and a higher molecular mass form of this toxin was purified from the extracellular products with the aim of inducing an immune response in fish. The optimal conditions for production of extracellular products by A. hydrophila strain B32 were studied to increase the concentration of this protoxin. The extracellular products were fractionated by molecular exclusion chromatography to yield a purified protoxin with an estimated molecular mass of 45 kDa by SDS-PAGE and which gave a positive reaction in Western blotting with the rabbit anti-15-kDa toxin serum. Since the 45-kDa protoxin showed lower specific acetylcholinesterase activity than the active 15-kDa toxin, the behaviour of the active site was studied using specific inhibitors. This 45-kDa protoxin was 13.3-fold less toxic than the 15-kDa toxin and induced antibody production in fish.

  13. Chitin degradation and electricity generation by Aeromonas hydrophila in microbial fuel cells.

    PubMed

    Li, Shan-Wei; He, Hui; Zeng, Raymond J; Sheng, Guo-Ping

    2017-02-01

    Chitin is one of the most abundant biopolymers in nature and the main composition of shrimp and crab shells (usually as food wastes). Thus it is essential to investigate the potential of degrading chitin for energy recovery. This study investigated the anaerobic degradation of chitin by Aeromonas hydrophila, a chitinolytic and popular electroactive bacterium, in both fermentation and microbial fuel cell (MFC) systems. The primary chitin metabolites produced in MFC were succinate, lactate, acetate, formate, and ethanol. The total metabolite concentration from chitin degradation increased seven-fold in MFC compared to the fermentation system, as well as additional electricity generation. Moreover, A. hydrophila degraded GlcNAc (the intermediate of chitin hydrolysis) significantly faster (0.97 and 0.94 mM C/d/mM-GlcNAc) than chitin (0.13 and 0.03 mM C/d/mM-GlcNAc) in MFC and fermentation systems, indicating that extracellular hydrolysis of chitin was the rate-limiting step and this step could be accelerated in MFC. Furthermore, more chemicals produced by the addition of exogenous mediators in MFC. This study proves that the chitin could be degraded effectively by an electroactive bacterium in MFC, and our results suggest that this bioelectrochemical system might be useful for the degradation of recalcitrant biomass to recover energy.

  14. Characterization of Aeromonas hydrophila Wound Pathotypes by Comparative Genomic and Functional Analyses of Virulence Genes

    PubMed Central

    Grim, Christopher J.; Kozlova, Elena V.; Sha, Jian; Fitts, Eric C.; van Lier, Christina J.; Kirtley, Michelle L.; Joseph, Sandeep J.; Read, Timothy D.; Burd, Eileen M.; Tall, Ben D.; Joseph, Sam W.; Horneman, Amy J.; Chopra, Ashok K.; Shak, Joshua R.

    2013-01-01

    ABSTRACT Aeromonas hydrophila has increasingly been implicated as a virulent and antibiotic-resistant etiologic agent in various human diseases. In a previously published case report, we described a subject with a polymicrobial wound infection that included a persistent and aggressive strain of A. hydrophila (E1), as well as a more antibiotic-resistant strain of A. hydrophila (E2). To better understand the differences between pathogenic and environmental strains of A. hydrophila, we conducted comparative genomic and functional analyses of virulence-associated genes of these two wound isolates (E1 and E2), the environmental type strain A. hydrophila ATCC 7966T, and four other isolates belonging to A. aquariorum, A. veronii, A. salmonicida, and A. caviae. Full-genome sequencing of strains E1 and E2 revealed extensive differences between the two and strain ATCC 7966T. The more persistent wound infection strain, E1, harbored coding sequences for a cytotoxic enterotoxin (Act), a type 3 secretion system (T3SS), flagella, hemolysins, and a homolog of exotoxin A found in Pseudomonas aeruginosa. Corresponding phenotypic analyses with A. hydrophila ATCC 7966T and SSU as reference strains demonstrated the functionality of these virulence genes, with strain E1 displaying enhanced swimming and swarming motility, lateral flagella on electron microscopy, the presence of T3SS effector AexU, and enhanced lethality in a mouse model of Aeromonas infection. By combining sequence-based analysis and functional assays, we characterized an A. hydrophila pathotype, exemplified by strain E1, that exhibited increased virulence in a mouse model of infection, likely because of encapsulation, enhanced motility, toxin secretion, and cellular toxicity. PMID:23611906

  15. Phytoextracts-Synthesized Silver Nanoparticles Inhibit Bacterial Fish Pathogen Aeromonas hydrophila.

    PubMed

    Mahanty, Arabinda; Mishra, Snehasish; Bosu, Ranadhir; Maurya, Uk; Netam, Surya Prakash; Sarkar, Biplab

    2013-12-01

    Fish disease is a major stumbling block towards sustainable growth of the fisheries sector. Aeromonas hydrophila, which is a major infectious aquatic pathogen is reportedly the causative agent of ulcers, fin-rot, tail-rot, hemorrhagic septicemia in fish, and has reportedly developed resistance against many of the available antibiotics. In this context, the inhibitory function of silver nanoparticles (AgNPs) against A. hydrophila was studied to evaluate its possible application in aquaculture as alternative to antibiotics. AgNPs were synthesized using the leaf extracts of subtropical plants Mangifera indica (Mango), Eucalyptus terticornis (Eucalyptus), Carica papaya (Papaya) and Musa paradisiaca (Banana). The absorbance maxima, size range and shape of the AgNPs as characterized by the UV-Vis spectroscopy, high resolution transmission electron microscopy (HR-TEM), and energy dispersive X-ray spectroscopy (EDX) were, Mangifera-442, 50-65 nm, ovular; Eucalyptus-465, 60-150 nm, oval; Carica-442, 25-40 nm, round, irregular; and Musa-454, 10-50 nm, round, irregular, respectively. Well-diffusion of these AgNPs for their antimicrobial characteristics exhibited that, the papaya leaf extract synthesized AgNPs had maximum antimicrobial activity at 153.6 μg/ml concentrations, and that from the eucalyptus leaves was least effective. As observed, the potency of the nanoparticles enhanced with the decrease in particle size, from 60-150 nm in eucalyptus to 25-40 nm in papaya. Due to its purely natural sourcing, phytosynthesized AgNPs can be applied as alternative to antibiotics and other biocides as a cost-effective and eco-friendly therapeutic agent against A. hydrophila stimulated diseases in aquatic animals.

  16. Mutation in the S-ribosylhomocysteinase (luxS) gene involved in quorum sensing affects biofilm formation and virulence in a clinical isolate of Aeromonas hydrophila

    EPA Science Inventory

    A diarrheal isolate SSU of Aeromonas hydrophila produces a cytotoxic enterotoxin (Act) with cytotoxic, enterotoxic, and hemolytic activities. Our laboratory has characterized from the above Aeromonas strain, in addition to Act, the type 3- and T6-secretion systems and their effec...

  17. A simple fluorogenic method to detect Vibrio cholerae and Aeromonas hydrophila in well water for areas impacted by catastrophic disasters.

    PubMed

    Richards, Gary P; Watson, Michael A

    2006-09-01

    The colony overlay procedure for peptidases (COPP) is a simple, fluorogenic assay that can rapidly detect and quantify Vibrio cholerae and Aeromonas hydrophila in well water. Cleavage of the substrate L-lysyl-7-amino-4-trifluoromethylcoumarin by enzymes present in Vibrio and Aeromonas species produces fluorescent foci on cellulose acetate membranes exposed to long-wave ultraviolet light. Vibrio cholerae O1, O139, O155, and A. hydrophila were readily detected using this procedure, whereas Enterobacteriaceae and other non-Vibrionaceae pathogens did not produce fluorescence. The assay is practical for assessing the relative safety of well water in areas that have experienced catastrophic devastation from natural disasters, acts of war, or civil strife and may help curb outbreaks of cholera and other enteric illnesses in affected areas. In tropical climates, the procedure may be adapted for use in areas without electricity.

  18. Functional Genomic Characterization of Virulence Factors from Necrotizing Fasciitis-Causing Strains of Aeromonas hydrophila

    PubMed Central

    Grim, Christopher J.; Kozlova, Elena V.; Ponnusamy, Duraisamy; Fitts, Eric C.; Sha, Jian; Kirtley, Michelle L.; van Lier, Christina J.; Tiner, Bethany L.; Erova, Tatiana E.; Joseph, Sandeep J.; Read, Timothy D.; Shak, Joshua R.; Joseph, Sam W.; Singletary, Ed; Felland, Tracy; Baze, Wallace B.; Horneman, Amy J.

    2014-01-01

    The genomes of 10 Aeromonas isolates identified and designated Aeromonas hydrophila WI, Riv3, and NF1 to NF4; A. dhakensis SSU; A. jandaei Riv2; and A. caviae NM22 and NM33 were sequenced and annotated. Isolates NF1 to NF4 were from a patient with necrotizing fasciitis (NF). Two environmental isolates (Riv2 and -3) were from the river water from which the NF patient acquired the infection. While isolates NF2 to NF4 were clonal, NF1 was genetically distinct. Outside the conserved core genomes of these 10 isolates, several unique genomic features were identified. The most virulent strains possessed one of the following four virulence factors or a combination of them: cytotoxic enterotoxin, exotoxin A, and type 3 and 6 secretion system effectors AexU and Hcp. In a septicemic-mouse model, SSU, NF1, and Riv2 were the most virulent, while NF2 was moderately virulent. These data correlated with high motility and biofilm formation by the former three isolates. Conversely, in a mouse model of intramuscular infection, NF2 was much more virulent than NF1. Isolates NF2, SSU, and Riv2 disseminated in high numbers from the muscular tissue to the visceral organs of mice, while NF1 reached the liver and spleen in relatively lower numbers on the basis of colony counting and tracking of bioluminescent strains in real time by in vivo imaging. Histopathologically, degeneration of myofibers with significant infiltration of polymorphonuclear cells due to the highly virulent strains was noted. Functional genomic analysis provided data that allowed us to correlate the highly infectious nature of Aeromonas pathotypes belonging to several different species with virulence signatures and their potential ability to cause NF. PMID:24795370

  19. The use of eugenol against Aeromonas hydrophila and its effect on hematological and immunological parameters in silver catfish (Rhamdia quelen).

    PubMed

    Sutili, Fernando Jonas; Kreutz, Luiz Carlos; Noro, Mirela; Gressler, Leticia Trevisan; Heinzmann, Berta Maria; de Vargas, Agueda Castagna; Baldisserotto, Bernardo

    2014-02-15

    The aim of this study was to evaluate the activity of eugenol against the fish pathogen Aeromonas hydrophila and eugenol's effect on hematological and natural immune parameters in silver catfish (Rhamdia quelen). In vitro, eugenol showed weak activity against A. hydrophila, but in vivo, at a subinhibitory concentration (10 mg L(-1)), it promoted survival in infected silver catfish. Eugenol (50 μg mL(-1)) reduced the hemolytic activity of A. hydrophila supernatant in vitro in fish erythrocytes. Subjecting catfish to eugenol baths (5 and 10 mg L(-1)) for five days did not alter the hematological and immunological parameters studied in this work. Based on these results, eugenol can be used to treat or prevent bacterial diseases in fish.

  20. Aloysia triphylla essential oil as additive in silver catfish diet: Blood response and resistance against Aeromonas hydrophila infection.

    PubMed

    Dos Santos, Alessandro C; Sutili, Fernando J; Heinzmann, Berta M; Cunha, Mauro A; Brusque, Isabel C M; Baldisserotto, Bernardo; Zeppenfeld, Carla C

    2017-03-01

    The essential oil of Aloysia triphylla (EOAT) is a promising product with potential use in aquaculture systems. This study evaluated hematological/biochemical responses and survival of silver catfish (Rhamdia quelen) fed a diet containing EOAT and infected by Aeromonas hydrophila. After 21 days of feeding trial, fish were infected with A. hydrophila following a 10-day period of observation. Blood collection was performed before and after the bacterial challenge. Dietary EOAT by itself seems to affect some blood parameters, decreasing total leukocyte, lymphocyte, and neutrophil counts and increasing total protein values. However, 2.0 mL EOAT/kg diet showed a possible potential protective effect after A. hydrophila infection, maintaining the evaluated parameters similar to basal values (from healthy fish before the feeding trial) and promoting survival of silver catfish.

  1. The FlgT Protein Is Involved in Aeromonas hydrophila Polar Flagella Stability and Not Affects Anchorage of Lateral Flagella

    PubMed Central

    Merino, Susana; Tomás, Juan M.

    2016-01-01

    Aeromonas hydrophila sodium-driven polar flagellum has a complex stator-motor. Consist of two sets of redundant and non-exchangeable proteins (PomA/PomB and PomA2/PomB2), which are homologs to other sodium-conducting polar flagellum stator motors; and also two essential proteins (MotX and MotY), that they interact with one of those two redundant pairs of proteins and form the T-ring. In this work, we described an essential protein for polar flagellum stability and rotation which is orthologs to Vibrio spp. FlgT and it is encoded outside of the A. hydrophila polar flagellum regions. The flgT was present in all mesophilic Aeromonas strains tested and also in the non-motile Aeromonas salmonicida. The A. hydrophila ΔflgT mutant is able to assemble the polar flagellum but is more unstable and released into the culture supernatant from the cell upon completion assembly. Presence of FlgT in purified polar hook-basal bodies (HBB) of wild-type strain was confirmed by Western blotting and electron microscopy observations showed an outer ring of the T-ring (H-ring) which is not present in the ΔflgT mutant. Anchoring and motility of proton-driven lateral flagella was not affected in the ΔflgT mutant and specific antibodies did not detect FlgT in purified lateral HBB of wild type strain. PMID:27507965

  2. Nutritional versatility and growth kinetics of an Aeromonas hydrophila strain isolated from drinking water.

    PubMed Central

    van der Kooij, D; Hijnen, W A

    1988-01-01

    The nutritional versatility and growth kinetics of Aeromonas hydrophila were studied to determine the nature and the growth-promoting properties of organic compounds which may serve as substrates for the growth of this organism in drinking water during treatment and distribution. As an initial screening, a total of 69 different organic compounds were tested at a concentration of 2.5 g/liter as growth substrates for 10 A. hydrophila strains. Of these strains, strain M800 attained the highest maximum colony counts in various types of drinking water and river water and was therefore used in further measurements of growth at low substrate concentrations. A mixture of 21 amino acids and a mixture of 10 long-chain fatty acids, when added to drinking water, promoted growth of strain M800 at individual compound concentrations as low as 0.1 microgram of C per liter. Mixtures of 18 carbohydrates and 18 carboxylic acids clearly enhanced growth of the organism at individual compound concentrations above 1 microgram of C per liter. Growth measurements with 63 individual substrates at a concentration of 10 micrograms of C per liter gave growth rates of greater than or equal to 0.1/h with two amino acids, nine carbohydrates, and six long-chain fatty acids. Ks values were determined for arginine (less than or equal to 0.3 micrograms of C per liter), glucose (15.9 micrograms of C per liter), acetate (11.1 micrograms of C per liter), and oleate (2.1 micrograms of C per liter). The data obtained indicate that biomass components, such as amino acids and long-chain fatty acids, can promote multiplication of aeromonads in drinking water distribution systems at concentrations as low as a few micrograms per liter. PMID:3214162

  3. Cross-Talk between the Aeromonas hydrophila Type III Secretion System and Lateral Flagella System

    PubMed Central

    Zhao, Yu-Hang; Shaw, Jonathan G.

    2016-01-01

    Aeromonas hydrophila is responsible for aeromonad septicaemia in fish, and gastroenteritis and wound infections in humans. The type III secretion system (T3SS) is utilized by aeromonads to inject protein effectors directly into host cells. One of the major genetic regulators of the T3SS in several bacterial species is the AraC-like protein ExsA. Previous studies have suggested a link between T3SS regulation and lateral flagella expression. The aim of this study was to determine the genetic regulation of the T3SS and its potential interaction with the lateral flagella system in A. hydrophila. To investigate the genes encoding the T3SS regulatory components exsA, exsD, exsC, and exsE were mutated and the activities of the T3SS promoters were measured in wild type and mutant backgrounds demonstrating a regulatory network. The Exs proteins were shown to interact with each other by BACTH assay and Far-Western Blot. The findings suggested a regulatory cascade in which ExsE was bound to the chaperone protein ExsC. When ExsC was free it sequestered the anti-activator ExsD thus stopping the inhibition of the T3SS master regulator ExsA allowing T3SS expression. The T3SS regulatory components were also shown to affect the expression of the lateral flagella system. The activities of the lateral flagella promoters were shown to be repressed by the absence of ExsD and ExsE, suggesting that the T3SS master regulator ExsA was a negative regulator of the lateral flagella system. PMID:27656180

  4. Ecofriendly biosorption of dyes and metals by bacterial biomass of Aeromonas hydrophila RC1.

    PubMed

    Busi, Siddhardha; Chatterjee, Rahul; Rajkumari, Jobina; Hnamte, Sairengpuii

    2016-03-01

    The ability of dried bacterial biomass in azo dye and heavy metal removal from aqueous solution was explored. Biosorption of three textile dyes, Eriochrome black T (EBT), Acid Red 26 (AR) and Trypan blue (TB) and heavy metals (Pb and Cr) by dried biomass of Aeromonas hydrophila RC1, was investigated in a batch system under various parameters such as dye concentration, contact time, concentration of biomass, pH, and temperature. The experimental results showed that the extent of biosorption for dyes increased with increase in initial concentration of dyes, biomass concentration, contact time, temperature and decreased with increase in pH. The experimental isotherms data were analyzed using Langmuir and Freundlich isotherm equations. The Langmuir model yielded good fit to the experimental data (R² approximately 0.794, 0.844 and 0.969 for the dyes, EBT, AR and TB, respectively) with maximum monolayer adsorption capacity of 58.8 mg g⁻¹ for AR. Similarly results were obtained for heavy metals and the data fit in Langmuir model (R² value of 0.849 and 0.787) with q(m) value of 40 mg g⁻¹ for Pb. The results fit in pseudo first order kinetics with removal upto 96.67 % for Pb. Involvement of the surface characteristics of the biomass in biosorption was studied using scanning electron micrographs, FTIR, EDX and XRD analysis. Thus, use ofA. hydrophila RC1 biomass can be extensively employed in water treatment plants in order to get desired water quality in the most economical way.

  5. Protective efficacy of recombinant hemolysin co-regulated protein (Hcp) of Aeromonas hydrophila in common carp (Cyprinus carpio).

    PubMed

    Wang, Nannan; Wu, Yafeng; Pang, Maoda; Liu, Jin; Lu, Chengping; Liu, Yongjie

    2015-10-01

    Motile aeromonad septicemia (MAS) caused by Aeromonas hydrophila is one of the common bacterial causes of fish mortalities. Prophylactic vaccination against this and other diseases is essential for continued growth of aquaculture. The type VI secretion system (T6SS) plays a crucial role in the virulence of A. hydrophila. The hemolysin co-regulated protein (Hcp) is an integral component of the T6SS apparatus and is considered a hallmark of T6SS function. Here, the T6SS effector Hcp was expressed and characterized, and its immunogenicity and protective efficacy were evaluated in common carp (Cyprinus carpio). Hcp secretion was found to be strongly induced by low temperature in A. hydrophila. Immunoblot analysis demonstrated that Hcp is conserved among A. hydrophila strains of different origins. The vaccination with recombinant Hcp resulted in an increased survival (46.67%) in common carp during a 10-day challenge time compared to non-vaccinated fish (7.14%). The vaccinated fish also showed the significantly increased levels of IgM antibody in serum and cytokines such as inerleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in kidney, spleen and gills. The recombinant Hcp shows promise as a vaccine candidate against A. hydrophila.

  6. Lethal dose and clinical signs of Aeromonas hydrophila in Arapaima gigas (Arapaimidae), the giant fish from Amazon.

    PubMed

    Dias, Marcia K R; Sampaio, Luciana S; Proietti-Junior, Aldo A; Yoshioka, Eliane T O; Rodrigues, Dália P; Rodriguez, Anselmo F R; Ribeiro, Ricardo A; Faria, Fernando S E D V; Ozório, Rodrigo O A; Tavares-Dias, Marcos

    2016-05-30

    Aeromonas hydrophila is causing substantial economic losses in world aquaculture. This study determined the tolerance limit (LD50-96h) of A. hydrophila in Arapaima gigas, and also investigated the clinical signs after intradermal inoculation. Arapaima gigas fingerlings were inoculated intraperitoneally with 0 (control), 1.0×10(5), 1.0×10(6), 1.0×10(7), 1.0×10(9) and 1.0×10(10)CFU/mL of A. hydrophila for the determination of LD50-96h, which was 1.8×10(8)CFU/mL. In another trial with intradermal inoculation of 1.8×10(8)CFU/mL A. hydrophila, there was a 91.6% of mortality between 8 and 23h, and several clinical signs were found. As follows: depigmentation in the tegument, lesions in the tail and fins, loss of balance, reduction of respiratory movements, hemorrhagic foci, necrotic hemorrhages in the kidney, liver and swim bladder, splenomegaly, ascites in the abdominal cavity and hyperemia, enlargement of the gall bladder, among other clinical signs observed. The results showed that A. gigas has a relative tolerance to A. hydrophila when compared to other Neotropical fish species.

  7. Growth of Aeromonas hydrophila in the whey cheeses Myzithra, Anthotyros, and Manouri during storage at 4 and 12 degrees C.

    PubMed

    Papageorgiou, Demetrios K; Melas, Dimitrios S; Abrahim, Amin; Angelidis, Apostolos S

    2006-02-01

    The fresh whey cheeses Myzithra, Anthotyros, and Manouri were inoculated with Aeromonas hydrophila strain NTCC 8049 (type strain) or with an A. hydrophila strain isolated from food (food isolate) at levels of 3.0 to 5.0 x 10(2) CFU/g of cheese and stored at 4 or 12 degrees C. Duplicate samples of cheeses were tested for levels of A. hydrophila and pH after up to 29 days of storage. At 4 degrees C, A. hydrophila grew in Myzithra and Anthotyros with a generation time of ca. 19 h, but no growth was observed in Manouri. In Myzithra, average maximum populations of 8.87 log CFU/g (type strain) and 8.79 log CFU/g (food isolate) were recorded after 20 and 22 days of storage at 4 degrees C, respectively. The average maximum populations observed in Anthotyros stored at 4 degrees C were 6.72 log CFU/g (food isolate) and 6.13 log CFU/g (type strain) and were observed after 15 and 16 days of storage, respectively. A. hydrophila grew rapidly and reached high numbers in cheeses stored at 12 degrees C. The average generation times were 3.7 and 3.9 h (Myzithra), 4.1 and 6.1 h (Anthotyros), and 8.0 and 9.2 h (Manouri) for the type strain and the food isolate, respectively. Among the different whey cheese trials, the highest A. hydrophila population recorded (10.13 log CFU/g) was in Myzithra that had been inoculated with the food isolate after 8 days of storage at 12 degrees C. To prevent A. hydrophila growth in whey cheeses, efforts must be focused on preventing postprocessing contamination and temperature abuse during transportation and storage.

  8. Implication of lateral genetic transfer in the emergence of Aeromonas hydrophila isolates of epidemic outbreaks in channel catfish.

    PubMed

    Hossain, Mohammad J; Waldbieser, Geoffrey C; Sun, Dawei; Capps, Nancy K; Hemstreet, William B; Carlisle, Kristen; Griffin, Matt J; Khoo, Lester; Goodwin, Andrew E; Sonstegard, Tad S; Schroeder, Steven; Hayden, Karl; Newton, Joseph C; Terhune, Jeffery S; Liles, Mark R

    2013-01-01

    To investigate the molecular basis of the emergence of Aeromonas hydrophila responsible for an epidemic outbreak of motile aeromonad septicemia of catfish in the Southeastern United States, we sequenced 11 A. hydrophila isolates that includes five reference and six recent epidemic isolates. Comparative genomics revealed that recent epidemic A. hydrophila isolates are highly clonal, whereas reference isolates are greatly diverse. We identified 55 epidemic-associated genetic regions with 313 predicted genes that are present in epidemic isolates but absent from reference isolates and 35% of these regions are located within genomic islands, suggesting their acquisition through lateral gene transfer. The epidemic-associated regions encode predicted prophage elements, pathogenicity islands, metabolic islands, fitness islands and genes of unknown functions, and 34 of the genes encoded in these regions were predicted as virulence factors. We found two pilus biogenesis gene clusters encoded within predicted pathogenicity islands. A functional metabolic island that encodes a complete pathway for myo-inositol catabolism was evident by the ability of epidemic A. hydrophila isolates to use myo-inositol as a sole carbon source. Testing of A. hydrophila field isolates found a consistent correlation between myo-inositol utilization as a sole carbon source and the presence of an epidemic-specific genetic marker. All epidemic isolates and one reference isolate shared a novel O-antigen cluster. Altogether we identified four different O-antigen biosynthesis gene clusters within the 11 sequenced A. hydrophila genomes. Our study reveals new insights into the evolutionary changes that have resulted in the emergence of recent epidemic A. hydrophila strains.

  9. Implication of Lateral Genetic Transfer in the Emergence of Aeromonas hydrophila Isolates of Epidemic Outbreaks in Channel Catfish

    PubMed Central

    Hossain, Mohammad J.; Waldbieser, Geoffrey C.; Sun, Dawei; Capps, Nancy K.; Hemstreet, William B.; Carlisle, Kristen; Griffin, Matt J.; Khoo, Lester; Goodwin, Andrew E.; Sonstegard, Tad S.; Schroeder, Steven; Hayden, Karl; Newton, Joseph C.; Terhune, Jeffery S.; Liles, Mark R.

    2013-01-01

    To investigate the molecular basis of the emergence of Aeromonas hydrophila responsible for an epidemic outbreak of motile aeromonad septicemia of catfish in the Southeastern United States, we sequenced 11 A. hydrophila isolates that includes five reference and six recent epidemic isolates. Comparative genomics revealed that recent epidemic A. hydrophila isolates are highly clonal, whereas reference isolates are greatly diverse. We identified 55 epidemic-associated genetic regions with 313 predicted genes that are present in epidemic isolates but absent from reference isolates and 35% of these regions are located within genomic islands, suggesting their acquisition through lateral gene transfer. The epidemic-associated regions encode predicted prophage elements, pathogenicity islands, metabolic islands, fitness islands and genes of unknown functions, and 34 of the genes encoded in these regions were predicted as virulence factors. We found two pilus biogenesis gene clusters encoded within predicted pathogenicity islands. A functional metabolic island that encodes a complete pathway for myo-inositol catabolism was evident by the ability of epidemic A. hydrophila isolates to use myo-inositol as a sole carbon source. Testing of A. hydrophila field isolates found a consistent correlation between myo-inositol utilization as a sole carbon source and the presence of an epidemic-specific genetic marker. All epidemic isolates and one reference isolate shared a novel O-antigen cluster. Altogether we identified four different O-antigen biosynthesis gene clusters within the 11 sequenced A. hydrophila genomes. Our study reveals new insights into the evolutionary changes that have resulted in the emergence of recent epidemic A. hydrophila strains. PMID:24278351

  10. Quorum sensing in Aeromonas hydrophila and Aeromonas salmonicida: identification of the LuxRI homologs AhyRI and AsaRI and their cognate N-acylhomoserine lactone signal molecules.

    PubMed Central

    Swift, S; Karlyshev, A V; Fish, L; Durant, E L; Winson, M K; Chhabra, S R; Williams, P; Macintyre, S; Stewart, G S

    1997-01-01

    Spent culture supernatants from both Aeromonas hydrophila and Aeromonas salmonicida activate a range of biosensors responsive to N-acylhomoserine lactones (AHLs). The genes for a quorum sensing signal generator and a response regulator were cloned from each Aeromonas species and termed ahyRI and asaRI, respectively. Protein sequence homology analysis places the gene products within the growing family of LuxRI homologs. ahyR and asaR are transcribed divergently from ahyI and asaI, respectively, and in both Aeromonas species, the genes downstream have been identified by DNA sequence and PCR analysis. Downstream of both ahyI and asaI is a gene with close homology to iciA, an inhibitor of chromosome replication in Escherichia coli, a finding which implies that in Aeromonas, cell division may be linked to quorum sensing. The major signal molecule synthesized via both AhyI and AsaI was purified from spent culture supernatants and identified as N-(butanoyl)-L-homoserine lactone (BHL) by thin-layer chromatography, high-pressure liquid chromatography analysis, and mass spectrometry. In addition, a second, minor AHL, N-hexanoyl-L-homoserine lactone, was identified. Transcriptional reporter studies with ahyI::luxCDABE fusions indicate that AhyR and BHL are both required for ahyI transcription. For A. salmonicida, although the addition of exogenous BHL gives only a small stimulation of the production of serine protease with comparison to the control culture, the incorporation of a longer-chain AHL, N-(3-oxodecanoyl)-L-homoserine lactone, reduced the final level (by approximately 50%) and delayed the appearance (from an A650 of 0.9 in the control to an A650 of 1.2 in the test) of protease in the culture supernatant. These data add A. hydrophila and A. salmonicida to the growing family of gram-negative bacteria now known to control gene expression through quorum sensing. PMID:9286976

  11. Thin-film fixed-bed reactor for solar photocatalytic inactivation of Aeromonas hydrophila: influence of water quality

    PubMed Central

    2012-01-01

    Background Controlling fish disease is one of the major concerns in contemporary aquaculture. The use of antibiotics or chemical disinfection cannot provide a healthy aquaculture system without residual effects. Water quality is also important in determining the success or failure of fish production. Several solar photocatalytic reactors have been used to treat drinking water or waste water without leaving chemical residues. This study has investigated the impact of several key aspects of water quality on the inactivation of the pathogenic bacterium Aeromonas hydrophila using a pilot-scale thin-film fixed-bed reactor (TFFBR) system. Results The level of inactivation of Aeromonas hydrophila ATCC 35654 was determined using a TFFBR with a photocatalytic area of 0.47 m2 under the influence of various water quality variables (pH, conductivity, turbidity and colour) under high solar irradiance conditions (980–1100 W m-2), at a flow rate of 4.8 L h-1 through the reactor. Bacterial enumeration were obtained through conventional plate count using trypticase soy agar media, cultured in conventional aerobic conditions to detect healthy cells and under ROS-neutralised conditions to detect both healthy and sub-lethally injured (oxygen-sensitive) cells. The results showed that turbidity has a major influence on solar photocatalytic inactivation of A. hydrophila. Humic acids appear to decrease TiO2 effectiveness under full sunlight and reduce microbial inactivation. pH in the range 7–9 and salinity both have no major effect on the extent of photoinactivation or sub-lethal injury. Conclusions This study demonstrates the effectiveness of the TFFBR in the inactivation of Aeromonas hydrophila under the influence of several water quality variables at high solar irradiance, providing an opportunity for the application of solar photocatalysis in aquaculture systems, as long as turbidity remains low. PMID:23194331

  12. Chemical modification of polyvinyl chloride and silicone elastomer in inhibiting adhesion of Aeromonas hydrophila.

    PubMed

    Kregiel, Dorota; Berlowska, Joanna; Mizerska, Urszula; Fortuniak, Witold; Chojnowski, Julian; Ambroziak, Wojciech

    2013-07-01

    Disease-causing bacteria of the genus Aeromonas are able to adhere to pipe materials, colonizing the surfaces and forming biofilms in water distribution systems. The aim of our research was to study how the modification of materials used commonly in the water industry can reduce bacterial cell attachment. Polyvinyl chloride and silicone elastomer surfaces were activated and modified with reactive organo-silanes by coupling or co-crosslinking silanes with the native material. Both the native and modified surfaces were tested using the bacterial strain Aeromonas hydrophila, which was isolated from the Polish water distribution system. The surface tension of both the native and modified surfaces was measured. To determine cell viability and bacterial adhesion two methods were used, namely plate count and luminometry. Results were expressed in colony-forming units (c.f.u.) and in relative light units (RLU) per cm(2). Almost all the chemically modified surfaces exhibited higher anti-adhesive and anti-microbial properties in comparison to the native surfaces. Among the modifying agents examined, poly[dimethylsiloxane-co-(N,N-dimethyl-N-n-octylammoniopropyl chloride) methylsiloxane)] terminated with hydroxydimethylsilyl groups (20 %) in silicone elastomer gave the most desirable results. The surface tension of this modifier, was comparable to the non-polar native surface. However, almost half of this value was due to the result of polar forces. In this case, in an adhesion analysis, only 1 RLU cm(-2) and less than 1 c.f.u. cm(-2) were noted. For the native gumosil, the results were 9,375 RLU cm(-2) and 2.5 × 10(8) c.f.u. cm(-2), respectively. The antibacterial activity of active organo-silanes was associated only with the carrier surface because no antibacterial compounds were detected in liquid culture media, in concentrations that were able to inhibit cell growth.

  13. Effect of Euphorbia hirta plant leaf extract on immunostimulant response of Aeromonas hydrophila infected Cyprinus carpio

    PubMed Central

    Sukumaran, NatarajaPillai

    2014-01-01

    The main objective of the present study is to improve the immune power of Cyprinus carpio by using Euphorbia hirta plant leaf extract as immunostimulants. The haematological, immunological and enzymatic studies were conducted on the medicated fish infected with Aeromonas hydrophila pathogen. The results obtained from the haematological studies show that the RBC count, WBC count and haemoglobin content were increased in the infected fish at higher concentration of leaf extract. The feeds with leaf extract of Euphorbia hirta were able to stimulate the specific immune response by increasing the titre value of antibody. It was able to stimulate the antibody production only up to the 5th day, when fed with higher concentrations of (25 g and 50 g) plant leaf extract. The plant extract showed non-specific immune responses such as lysozyme activity, phagocytic ratio, NBT assay, etc. at higher concentration (50 g) and in the same concentration (50 g), the leaf extract of Euphorbia hirta significantly eliminated the pathogen in blood and kidney. It was observed that fish have survival percentage significantly at higher concentration (50 g) of Euphorbia hirta, when compared with the control. The obtained results are statistically significant at P < 0.05 and P < 0.01 levels. This research work suggests that the plant Euphorbia hirta has immunostimulant activity by stimulating both specific and non-specific immunity at higher concentrations. PMID:25405077

  14. Extractive biodecolorization of triphenylmethane dyes in cloud point system by Aeromonas hydrophila DN322p.

    PubMed

    Pan, Tao; Ren, Suizhou; Xu, Meiying; Sun, Guoping; Guo, Jun

    2013-07-01

    The biological treatment of triphenylmethane dyes is an important issue. Most microbes have limited practical application because they cannot completely detoxicate these dyes. In this study, the extractive biodecolorization of triphenylmethane dyes by Aeromonas hydrophila DN322p was carried out by introducing the cloud point system. The cloud point system is composed of a mixture of nonionic surfactants (20 g/L) Brij 30 and Tergitol TMN-3 in equal proportions. After the decolorization of crystal violet, a higher wet cell weight was obtained in the cloud point system than that of the control system. Based on the results of thin-layer chromatography, the residual crystal violet and its decolorized product, leuco crystal violet, preferred to partition into the coacervate phase. Therefore, the detoxification of the dilute phase was achieved, which indicated that the dilute phase could be discharged without causing dye pollution. The extractive biodecolorization of three other triphenylmethane dyes was also examined in this system. The decolorization of malachite green and brilliant green was similar to that of crystal violet. Only ethyl violet achieved a poor decolorization rate because DN322p decolorized it via adsorption but did not convert it into its leuco form. This study provides potential application of biological treatment in triphenylmethane dye wastewater.

  15. Effect of plasma processing and organosilane modifications of polyethylene on Aeromonas hydrophila biofilm formation.

    PubMed

    Kregiel, Dorota; Niedzielska, Kamila

    2014-01-01

    The aim of our research was to study how the modifications of polyethylene--a material commonly used in medicine and water industry--influence bacterial cell attachment and biofilm formation. The native surface was activated and modified using two-step process consisting in the activation of native surface with a H2O vapor plasma followed by its treatment with various organosilanes, namely, [3(tertbutylamine-2hydroxy) propyloxypropyl] diethoxymethylsilane, 1H,1H,2H,2H-perfluorooctylmethyldimethoxysilane, dimethoxydimethylsilane, and isobutylmethyldimethoxysilane. The effect of polyethylene modification after chemical treatment was analyzed using surface tension measurement. The adhesive properties of Aeromonas hydrophila LOCK0968 were studied in water with a low concentration of organic compounds, using luminometric and microscopic methods, and the viability of the adhered bacterial cells was evaluated using the colony forming units method. After two-week incubation the chemically modified materials exhibited better antiadhesive and antibacterial characteristics in comparison to the native surface. Among the examined modifying agents, dimethoxydimethylsilane showed the best desired properties.

  16. Anti-biofouling property of vanillin on Aeromonas hydrophila initial biofilm on various membrane surfaces.

    PubMed

    Ponnusamy, K; Kappachery, S; Thekeettle, M; Song, J H; Kweon, J H

    2013-09-01

    Biofouling is a serious problem on filter membranes of water purification systems due to formation of bacterial biofilms, which can be detrimental to the membrane performance. Biofouling occurs on membrane surface and therefore greatly influences the physical and chemical aspects of the surface. Several membranes including microfiltration, ultrafiltration, and reverse osmosis (RO) membranes were used to learn about the anti-biofouling properties of vanillin affecting the membrane performances. Vanillin has been recognized as a potential quorum quenching compound for Aeromonas hydrophila biofilms. The initial attachment and dynamics of biofilm growth were monitored using scanning electron microscopy and confocal laser scanning microscopy. Biofilm quantities were measured using a plate count method and total protein determinations. Vanillin addition was effective in the prevention of biofilm formation on the tested membrane surfaces. Among the membranes, RO membranes made with cellulose acetate showed the most substantial reduction of biofilm formation by addition of vanillin. The biofilm reduction was confirmed by the results of surface coverage, biomass and protein accumulation. The HPLC spectrum of the spent culture with vanillin addition showed that vanillin may interfere with quorum sensing molecules and thus prevent the formation of the biofilms.

  17. Structural Insights on PHA Binding Protein PhaP from Aeromonas hydrophila

    PubMed Central

    Zhao, Hongyu; Wei, Hui; Liu, Xi; Yao, Zhenyu; Xu, Manyu; Wei, Daixu; Wang, Jiawei; Wang, Xinquan; Chen, Guo-Qiang

    2016-01-01

    Phasins or PhaPs are a group of amphiphilic proteins that are found attached to the surface of microbial polyhydroxyalkanoate (PHA) granules. They have both structural and regulatory functions and can affect intracellular PHA accumulation and mediate protein folding. The molecular basis for the diverse functions of the PhaPs has not been fully understood due to the lack of the structural knowledge. Here we report the structural and biochemical studies of the PhaP cloned from Aeromonas hydrophila (PhaPAh), which is utilized in protein and tissue engineering. The crystal structure of PhaPAh was revealed to be a tetramer with 8 α-helices adopting a coiled-coil structure. Each monomer has a hydrophobic and a hydrophilic surface, rendering the surfactant properties of the PhaPAh monomer. Based on the crystal structure, we predicted three key amino acid residues and obtained mutants with enhanced stability and improved emulsification properties. The first PhaP crystal structure, as reported in this study, is an important step towards a mechanistic understanding of how PHA is formed in vivo and why PhaP has such unique surfactant properties. At the same time, it will facilitate the study of other PhaP members that may have significant biotechnological potential as bio-surfactants and amphipathic coatings. PMID:28009010

  18. A tandem-repeat galectin-9 involved in immune response of yellow catfish, Pelteobagrus fulvidraco, against Aeromonas hydrophila.

    PubMed

    Wang, Yun; Ke, Fei; Ma, Jingjing; Zhou, Shuaibang

    2016-04-01

    Galectins exclusively recognize and bind β-galactoside on cell surface by carbohydrate recognition domain (CRD). In spite of extensive study of mammalian galectin importance in immune system, little is known about that of fish. To study the immune response of yellow catfish to pathogens, a tandem-repeat galectin-9 from yellow catfish was identified and named PfGAL9. Its full-length cDNA was 1314 bp, including a 117 bp of 5' untranslated region (UTR), a 951 bp of open reading frame (ORF), and a 246 bp of 3' UTR. The ORF encoded 316 amino acids (35.12 KDa), shared the highest 78% identity with the predicted galectin-9 of Ictalurus punctatus. This protein possessed two distinct CRDs with two highly conserved sugar binding motifs. Quantitative PCR showed that PfGAL9 was lowly expressed in skin, gill, fin, muscle, heart, and intestine, highly expressed in tested immune tissues (head kidney, trunk kidney, liver, spleen, and blood) in normal body. After inactivated Aeromonas hydrophila challenge, PfGAL9 was remarkably increased in head kidney and liver in a time-dependent manner. The recombinant protein was expressed in Escherichia coli, which not only agglutinated but also bond all examined bacteria. The binding activities are consistent with the size of aggregates formed by agglutinated bacteria. The agglutination must depend on its direct interaction with bacteria. These results suggested that PfGAL9 was involved in the innate immune response against bacterial infection and clearance of pathogens in yellow catfish.

  19. A study on genetic variability of pathogenic Aeromonas hydrophila strains and the varied responses of the strains towards phyto-extracts.

    PubMed

    Balasundaram, A; Kumari, P Rathna; Kolanchinathan, P; Masilamani, V; John, George

    2013-11-01

    The present study evaluated genetic variation in Aeromonas hydrophila strains using PCR-RAPD and their varied susceptibility to phyto-extract. Four strains of Aeromonas hydrophila isolated from skin infections of common freshwater fish, Cyprinus carpio were characterized by various biochemical methods, physiological tests and PCR- RAPD. Antimicrobial activity of the leaf extracts of three medicinal plants, Ocimum sanctum, Adathoda vasica and Calendula officinalis were tested against the four strains of A. hydrophila by disc diffusion (Kirby-Bauer) method. Antagonistic effects of leaf extracts against A. hydrophila strains were assessed by co-culture method. RAPD analysis showed that all the microbes isolated from skin infection belong to the same species but there was no 100% genetic similarity among them Dendrogram constructed by UPGMA clearly supported the PCR pattern of genetic variability among the strains. This study revealed that Aeromonas hydophila exhibits genetic variability and varied susceptibility towards phyto-extracts. Results indicated that phyto-extracts offers a promising alternative to the use of antibiotics in controlling Aeromonas hydrophila.

  20. Identification of Aeromonas hydrophila Genes Preferentially Expressed after Phagocytosis by Tetrahymena and Involvement of Methionine Sulfoxide Reductases.

    PubMed

    Pang, Maoda; Lin, Xiaoqin; Liu, Jin; Guo, Changming; Gao, Shanshan; Du, Hechao; Lu, Chengping; Liu, Yongjie

    2016-01-01

    Free-living protozoa affect the survival and virulence evolution of pathogens in the environment. In this study, we explored the fate of Aeromonas hydrophila when co-cultured with the bacteriovorous ciliate Tetrahymena thermophila and investigated bacterial gene expression associated with the co-culture. Virulent A. hydrophila strains were found to have ability to evade digestion in the vacuoles of this protozoan. In A. hydrophila, a total of 116 genes were identified as up-regulated following co-culture with T. thermophila by selective capture of transcribed sequences (SCOTS) and comparative dot-blot analysis. A large proportion of these genes (42/116) play a role in metabolism, and some of the genes have previously been characterized as required for bacterial survival and replication within macrophages. Then, we inactivated the genes encoding methionine sulfoxide reductases, msrA, and msrB, in A. hydrophila. Compared to the wild-type, the mutants ΔmsrA and ΔmsrAB displayed significantly reduced resistance to predation by T. thermophila, and 50% lethal dose (LD50) determinations in zebrafish demonstrated that both mutants were highly attenuated. This study forms a solid foundation for the study of mechanisms and implications of bacterial defenses.

  1. A transcriptome analysis focusing on inflammation-related genes of grass carp intestines following infection with Aeromonas hydrophila

    PubMed Central

    Song, Xuehong; Hu, Xiaolong; Sun, Bingyao; Bo, Yunxuan; Wu, Kang; Xiao, Lanying; Gong, Chengliang

    2017-01-01

    Inflammation is a protective response that is implicated in bacterial enteritis and other fish diseases. The inflammatory mechanisms behind Aeromonas hydrophila infections in fish remain poorly understood. In this study, we performed a de novo grass carp transcriptome assembly using Illumina’s Solexa sequencing technique. On this basis we carried out a comparative analysis of intestinal transcriptomes from A. hydrophila-challenged and physiological saline solution (PSS/mock) -challenged fish, and 315 genes were up-regulated and 234 were down-regulated in the intestines infected with A. hydrophila. The GO enrichment analysis indicated that the differentially expressed genes were enriched to 12, 4, and 8 GO terms in biological process, molecular function, and cellular component, respectively. A KEGG analysis showed that 549 DEGs were involved in 165 pathways. Moreover, 15 DEGs were selected for quantitative real-time PCR analysis to validate the RNA-seq data. The results confirmed the consistency of the expression levels between RNA-seq and qPCR data. In addition, a time-course analysis of the mRNA expression of 12 inflammatory genes further demonstrated that the intestinal inflammatory responses to A. hydrophila infection simultaneously modulated gene expression variations. The present study provides intestine-specific transcriptome data, allowing us to unravel the mechanisms of intestinal inflammation triggered by bacterial pathogens. PMID:28094307

  2. Identification and Characterization of MicroRNAs in the Liver of Blunt Snout Bream (Megalobrama amblycephala) Infected by Aeromonas hydrophila

    PubMed Central

    Cui, Lei; Hu, Hongtao; Wei, Wei; Wang, Weimin; Liu, Hong

    2016-01-01

    MicroRNAs (miRNAs) are small RNA molecules that play key roles in regulation of various biological processes. In order to better understand the biological significance of miRNAs in the context of Aeromonas hydrophila infection in Megalobrama amblycephala, small RNA libraries obtained from fish liver at 0 (non-infection), 4, and 24 h post infection (poi) were sequenced using Illumina deep sequencing technology. A total of 11,244,207, 9,212,958, and 7,939,157 clean reads were obtained from these three RNA libraries, respectively. Bioinformatics analysis identified 171 conserved miRNAs and 62 putative novel miRNAs. The existence of ten randomly selected novel miRNAs was validated by RT-PCR. Pairwise comparison suggested that 61 and 44 miRNAs were differentially expressed at 4 and 24 h poi, respectively. Furthermore, the expression profiles of nine randomly selected miRNAs were validated by qRT-PCR. MicroRNA target prediction, gene ontology (GO) annotation, and Kyoto Encylopedia of Genes and Genomes (KEGG) analysis indicated that a variety of biological pathways could be affected by A. hydrophila infection. Additionally, transferrin (TF) and transferrin receptor (TFR) genes were confirmed to be direct targets of miR-375. These results will expand our knowledge of the role of miRNAs in the immune response of M. amblycephala to A. hydrophila infection, and facilitate the development of effective strategies against A. hydrophila infection in M. amblycephala. PMID:27898025

  3. Identification of Aeromonas hydrophila Genes Preferentially Expressed after Phagocytosis by Tetrahymena and Involvement of Methionine Sulfoxide Reductases

    PubMed Central

    Pang, Maoda; Lin, Xiaoqin; Liu, Jin; Guo, Changming; Gao, Shanshan; Du, Hechao; Lu, Chengping; Liu, Yongjie

    2016-01-01

    Free-living protozoa affect the survival and virulence evolution of pathogens in the environment. In this study, we explored the fate of Aeromonas hydrophila when co-cultured with the bacteriovorous ciliate Tetrahymena thermophila and investigated bacterial gene expression associated with the co-culture. Virulent A. hydrophila strains were found to have ability to evade digestion in the vacuoles of this protozoan. In A. hydrophila, a total of 116 genes were identified as up-regulated following co-culture with T. thermophila by selective capture of transcribed sequences (SCOTS) and comparative dot-blot analysis. A large proportion of these genes (42/116) play a role in metabolism, and some of the genes have previously been characterized as required for bacterial survival and replication within macrophages. Then, we inactivated the genes encoding methionine sulfoxide reductases, msrA, and msrB, in A. hydrophila. Compared to the wild-type, the mutants ΔmsrA and ΔmsrAB displayed significantly reduced resistance to predation by T. thermophila, and 50% lethal dose (LD50) determinations in zebrafish demonstrated that both mutants were highly attenuated. This study forms a solid foundation for the study of mechanisms and implications of bacterial defenses. PMID:28083518

  4. Aeromonas hydrophila isolated from food and drinking water: hemagglutination, hemolysis, and cytotoxicity for a human intestinal cell line (HT-29).

    PubMed Central

    Handfield, M; Simard, P; Couillard, M; Letarte, R

    1996-01-01

    Aeromonas hydrophila isolated from food and drinking water was tested for pathogenicity by studying its hemolysis, hemagglutination, and cytotoxicity. Hemolysis, tested on erythrocytes from six different species, was more frequently seen with water isolates (64%) than with food isolates (48%). Hemagglutination was more frequently encountered with food isolates (92%) than with water isolates (73%). Cytotoxicity, evaluated on seven cell lines, was frequently observed with food isolates (92%) and with water isolates (73%). Heat treatment (56 degrees C for 10 min) of culture supernatant fluids inhibited the toxicity of some but not all toxin-producing isolates. Our results suggest that the human intestinal cell line HT-29 could be a useful complement for testing A. hydrophila exotoxins and for studying the enteropathogenicity of this species for humans. PMID:8795237

  5. Role of Aeromonas hydrophila flagella glycosylation in adhesion to Hep-2 cells, biofilm formation and immune stimulation.

    PubMed

    Merino, Susana; Wilhelms, Markus; Tomás, Juan M

    2014-11-28

    Polar flagellin proteins from Aeromonas hydrophila strain AH-3 (serotype O34) were found to be O-glycosylated with a heterogeneous heptasaccharide glycan. Two mutants with altered (light and strong) polar flagella glycosylation still able to produce flagella were previously obtained, as well as mutants lacking the O34-antigen lipopolysaccharide (LPS) but with unaltered polar flagella glycosylation. We compared these mutants, altogether with the wild type strain, in different studies to conclude that polar flagella glycosylation is extremely important for A. hydrophila adhesion to Hep-2 cells and biofilm formation. Furthermore, the polar flagella glycosylation is an important factor for the immune stimulation of IL-8 production via toll receptor 5 (TLR5).

  6. Protective efficacy of PLGA microspheres loaded with divalent DNA vaccine encoding the ompA gene of Aeromonas veronii and the hly gene of Aeromonas hydrophila in mice.

    PubMed

    Gao, Shanshan; Zhao, Na; Amer, Said; Qian, Mingming; Lv, Mengxi; Zhao, Yuliang; Su, Xin; Cao, Jieying; He, Hongxuan; Zhao, Baohua

    2013-11-19

    In the present study, poly (lactic-co-glycolic) acid (PLGA) was used as a carrier for a divalent fusion DNA vaccine encoding the Aeromonas veronii outer membrane protein A (ompA) and Aeromonas hydrophila hemolysins (hly) protein. The recombinant pET-28a-ompA-hly was constructed by inserting the ompA gene and hly gene into a pET-28a expression vector. Loading of ompA-hly antigen module on PLGA microspheres were accomplished by water-in-oil-in-water (W/O/W) encapsulation. The molecular weight and specificity of pET-28a-ompA-hly were detected by dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. The microspheres showed an average particle size of 100-150 μm and a loading efficiency (LE) of 68.8%. Mice received ompA-hly antigen-loaded PLGA microspheres by intraperitoneal or intragastric administration mounted strong and sustained IgG response, which was significantly higher (p<0.05) than those achieved by pET-28a-ompA-hly antigen alone. OmpA-hly antigen-loaded PLGA microsphere vaccine uniquely conferred a long lasting (30 days) sterile immunity against challenge infection. Results indicated that ompA-hly antigen-loaded PLGA microsphere vaccine is a qualified candidate vector system for sterile protective immunity against A. hydrophila and A. veronii infections.

  7. Skin and subcutaneous mycoses in tilapia (Oreochromis niloticus) caused by Fusarium oxysporum in coinfection with Aeromonas hydrophila.

    PubMed

    Cutuli, M Teresa; Gibello, Alicia; Rodriguez-Bertos, Antonio; Blanco, M Mar; Villarroel, Morris; Giraldo, Alejandra; Guarro, Josep

    2015-09-01

    Subcutaneous mycoses in freshwater fish are rare infections usually caused by oomycetes of the genus Saprolegnia and some filamentous fungi. To date, Fusarium infections in farmed fish have only been described in marine fish. Here, we report the presence of Fusarium oxysporum in subcutaneous lesions of Nile tilapia (Oreochromis niloticus). Histopathologic evaluation revealed granuloma formation with fungal structures, and the identity of the etiological agent was demonstrated by morphological and molecular analyses. Some of the animals died as a result of systemic coinfection with Aeromonas hydrophila.

  8. Immune effects of the vaccine of live attenuated Aeromonas hydrophila screened by rifampicin on common carp (Cyprinus carpio L).

    PubMed

    Jiang, Xinyu; Zhang, Chao; Zhao, Yanjing; Kong, Xianghui; Pei, Chao; Li, Li; Nie, Guoxing; Li, Xuejun

    2016-06-08

    Aeromonas hydrophila, as a strong Gram-negative bacterium, can infect a wide range of freshwater fish, including common carp Cyprinus carpio, and cause the huge economic loss. To create the effective vaccine is the best way to control the outbreak of the disease caused by A. hydrophila. In this study, a live attenuated A. hydrophila strain, XX1LA, was screened from the pathogenic A. hydrophila strain XX1 cultured on medium containing the antibiotic rifampicin, which was used as a live attenuated vaccine candidate. The immune protection of XX1LA against the pathogen A. hydrophila in common carp was evaluated by the relative percent survival (RPS), the specific IgM antibody titers, serum lysozyme activity and the expression profiles of multiple immune-related genes at the different time points following immunization. The results showed that the variable up-regulations of the immune-related genes, such as the pro-inflammatory cytokine IL-1β, the chemokine IL-10 and IgM, were observed in spleen and liver of common carp injected in the vaccines with the formalin-killed A. hydrophila (FKA) and the live attenuated XX1LA. Specific antibody to A. hydrophila was found to gradually increase during 28 days post-vaccination (dpv), and the RPS (83.7%) in fish vaccinated with XX1LA, was significant higher than that (37.2%) in fish vaccinated with FKA (P<0.05) on Day 28 after challenged by pathogen. It was demonstrated that the remarkable immune protection presented in the group vaccinated with XX1LA. During the late stage of 4-week immunization phase, compared with FKA and the control, specific IgM antibody titers significantly increased (P<0.05) in the XX1LA group. The activity of the lysozyme in serum indicated no significant change among three groups. In summary, the live attenuated bacterial vaccine XX1LA, screened in this study, indicates the better protect effect on common carp against A. hydrophila, which can be applied in aquaculture of common carp to prevent from the

  9. Proteomics Analysis Reveals a Potential Antibiotic Cocktail Therapy Strategy for Aeromonas hydrophila Infection in Biofilm.

    PubMed

    Li, Wanxin; Yao, Zujie; Sun, Lina; Hu, Wenjie; Cao, Jijuan; Lin, Wenxiong; Lin, Xiangmin

    2016-06-03

    Antibiotic fitness and acquired resistance are the two critical factors when bacteria respond to antibiotics, and the correlations and mechanisms between these two factors remain largely unknown. In this study, a TMT-labeling-based quantitative proteomics method was used to compare the differential expression of proteins between the fitness and acquired resistance to chlortetracycline in Aeromonas hydrophila biofilm. Bioinformatics analysis showed that translation-related ribosomal proteins, such as 30s ribosome subunits, increased in both factors; fatty acid biosynthesis related proteins, such as FabB, FabD, FabG, AccA, and AccD, increased in biofilm fitness, and some pathways (including propanoate-metabolism-related protein, such as PrpB, AtoB, PflB, AcsA, PrpD, and GabT) displayed decreased abundance in acquired resistance biofilm. The varieties of selected proteins involved in fatty acid biosynthesis and propanoate metabolism were further validated by q-PCR assay or Western blotting. Furthermore, the antibiotic-resistance-function assays showed that fatty-acid biosynthesis should be a protective antibiotics-resistance mechanism and a cocktail of chlortetracycline and triclosan, a fatty-acid-biosynthesis inhibitor, exhibited more efficient antimicrobial capability than did each antibiotic individually on biofilm, specifically on chlortetracycline-sensitive biofilm. We therefore demonstrate that the up-regulation of fatty acid biosynthesis may play an important role in antibiotic resistance and suggest that a cocktail of chlortetracycline and triclosan may be a potential cocktail therapy for pathogenic infections in biofilm.

  10. White Mulberry (Morus alba) Foliage Methanolic Extract Can Alleviate Aeromonas hydrophila Infection in African Catfish (Clarias gariepinus)

    PubMed Central

    Sheikhlar, Atefeh; Alimon, Abd Razk; Daud, Hassan; Saad, Chee R.; Webster, Carl D.; Meng, Goh Yong

    2014-01-01

    Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish. PMID:25574488

  11. Bacillus sp. QSI-1 Modulate Quorum Sensing Signals Reduce Aeromonas hydrophila Level and Alter Gut Microbial Community Structure in Fish.

    PubMed

    Zhou, Shuxin; Zhang, An; Yin, Hongping; Chu, Weihua

    2016-01-01

    Quorum sensing (QS) is a cell density dependent process that enables bacteria to communicate with each other based on the production, secretion and sensing of the auto-inducer molecules and then subsequently regulate virulence associated gene expression. Interrupting quorum sensing may represent a novel alternative approach to combat bacterial pathogen. Several bacteria can produce quorum quenching (QQ) enzymes. However, the role of QQ bacteria in shaping the microbiota and the level of N-acyl-homoserine lactones (AHLs, a prevalent type of QS molecules) producing bacteria remains largely unknown. The objective of this study was to examine the presence of AHLs in the fish intestine and investigate the modulation of gut microbiota and its effect on Aeromonas hydrophila level by a QQ enzyme producing probiotic Bacillus sp. QSI-1. AHLs were found in fish gut content and were confirmed in Aeromonas species using Chromobacterium violaceum CV026 and Agrobacterium tumefaciens AT 136 (pZLR4) as reporter strains. We demonstrated that the composition of fish gut microbiota was affected by quenching bacteria QSI-1, and the percentage of A. hydrophila was decreased significantly. Taken together, these results provide valuable insights into QQ enzyme producing probiotics can modulate the microbiota structure and decrease the percentage of AHL-producing pathogenic bacteria in the gut. These data strongly suggest that QQ probiotics may serve as non-antibiotic feed additive in aquaculture to control bacterial diseases.

  12. White mulberry (Morus alba) foliage methanolic extract can alleviate Aeromonas hydrophila infection in African catfish (Clarias gariepinus).

    PubMed

    Sheikhlar, Atefeh; Alimon, Abd Razk; Daud, Hassan; Saad, Chee R; Webster, Carl D; Meng, Goh Yong; Ebrahimi, Mahdi

    2014-01-01

    Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish.

  13. Bacillus sp. QSI-1 Modulate Quorum Sensing Signals Reduce Aeromonas hydrophila Level and Alter Gut Microbial Community Structure in Fish

    PubMed Central

    Zhou, Shuxin; Zhang, An; Yin, Hongping; Chu, Weihua

    2016-01-01

    Quorum sensing (QS) is a cell density dependent process that enables bacteria to communicate with each other based on the production, secretion and sensing of the auto-inducer molecules and then subsequently regulate virulence associated gene expression. Interrupting quorum sensing may represent a novel alternative approach to combat bacterial pathogen. Several bacteria can produce quorum quenching (QQ) enzymes. However, the role of QQ bacteria in shaping the microbiota and the level of N-acyl-homoserine lactones (AHLs, a prevalent type of QS molecules) producing bacteria remains largely unknown. The objective of this study was to examine the presence of AHLs in the fish intestine and investigate the modulation of gut microbiota and its effect on Aeromonas hydrophila level by a QQ enzyme producing probiotic Bacillus sp. QSI-1. AHLs were found in fish gut content and were confirmed in Aeromonas species using Chromobacterium violaceum CV026 and Agrobacterium tumefaciens AT 136 (pZLR4) as reporter strains. We demonstrated that the composition of fish gut microbiota was affected by quenching bacteria QSI-1, and the percentage of A. hydrophila was decreased significantly. Taken together, these results provide valuable insights into QQ enzyme producing probiotics can modulate the microbiota structure and decrease the percentage of AHL-producing pathogenic bacteria in the gut. These data strongly suggest that QQ probiotics may serve as non-antibiotic feed additive in aquaculture to control bacterial diseases. PMID:28018866

  14. Comparative transcriptional analysis reveals distinct expression patterns of channel catfish genes after the first infection and re-infection with Aeromonas hydrophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To determine whether transcriptional levels of channel catfish (Ictalurus punctatus) genes are differentially regulated between a first infection with Aeromonas hydrophila and a re-infection, suppression subtractive hybridization (SSH) was performed in this study using anterior kidney cDNA after the...

  15. Identification, virulence, and mass spectrometry of toxic ECP fractions of West Alabama isolates of Aeromonas hydrophila obtained from a 2010 disease outbreak

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In West Alabama, disease outbreaks in 2009 caused by Aeromonas hydrophila have led to an estimated loss of more than $3 million. In 2010, disease outbreak occurred again in West Alabama, causing losses of hundreds of thousands of pounds of market size channel catfish. During the 2010 disease outbrea...

  16. A Case of Extrusion of a Solid Silicone Tire Migrating through the Superior Rectus Muscle with Aeromonas hydrophila Infection following a Scleral Buckling Procedure

    PubMed Central

    Makino, Shinji; Sato, Yukihiro

    2012-01-01

    To our knowledge, there are no reports of Aeromonas hydrophila infection after a scleral buckling procedure. Also, migration of a silicone explant element through the rectus muscles is extremely rare. Herein, we describe a case experiencing extrusion of a solid silicone tire migrating through the superior rectus muscle with Aeromonas hydrophila infection following a scleral buckling procedure. A 42-year-old man was referred to our hospital complaining of ocular pain and purulent discharge in his left eye which has persisted for several months. He had a history of bilateral rhegmatogenous retinal detachment which had been treated with scleral buckling. The left eye showed extrusion of the solid silicone buckle which had migrated through the superior rectus muscle and an infection in the upper quadrant of the sclera. The buckle was removed, and the patient was treated with antibiotics. After the removal of the buckle, the symptoms showed rapid amelioration and there was no recurrence of retinal detachment. Aeromonas hydrophila was isolated from the discharge and the removed explant. He used well water in daily life. In this case, the Aeromonas hydrophila infection of the extruded buckle might have originated from contaminated well water. PMID:23213585

  17. Effect of Mangifera indica kernel as a feed additive on immunity and resistance to Aeromonas hydrophila in Labeo rohita fingerlings.

    PubMed

    Sahu, Swagatika; Das, Basanta Kumar; Pradhan, Jyotirmayee; Mohapatra, B C; Mishra, B K; Sarangi, Niranjan

    2007-07-01

    The study evaluated the efficacy of dietary doses of Mangifera indica (mango) kernel on the immune response and disease resistance of Labeo rohita fingerlings against the bacterial pathogen Aeromonas hydrophila infections in. L. rohita fingerlings fed diet containing 0 (Control), 1g, 5 g, 10 g mango kernel kg(-1) dry diet for 60 days. Biochemical (serum total protein, albumin, globulin, albumin:globulin ratio, blood glucose), haematological (WBC, RBC, haemoglobin content) and immunological (superoxide anion production, lysozyme, serum bactericidal activity) parameters of fish were examined at 20, 40 and 60 days of feeding. Fish were challenged with A. hydrophila 60 days post feeding and mortalities were recorded over 10 days post-infection. The results demonstrate that fish fed with mango kernel showed enhanced superoxide anion production, lysozyme, serum bactericidal, serum protein, albumin (P<0.05) compared with the control group. The mortality (%) was recorded up to 10 th day post-challenge. Less survivability was observed in control group (50%) up to day 10 after infection. The survivability was higher in experimental diets. The group fed 5 g kernel kg(-1) dry diet showed highest percentage survival (98%). These results indicate that mango kernel stimulates the immunity and makes L. rohita more resistant to A. hydrophila infection.

  18. Ameliorating ER-stress attenuates Aeromonas hydrophila-induced mitochondrial dysfunctioning and caspase mediated HKM apoptosis in Clarias batrachus

    PubMed Central

    Banerjee, Chaitali; Singh, Ambika; Das, Taposh Kumar; Raman, Rajagopal; Shrivastava, Anju; Mazumder, Shibnath

    2014-01-01

    Endoplasmic reticulum (ER)-stress and unfolding protein response (UPR) has not been implied in Aeromonas hydrophila-pathogenicity. We report increased expression of the ER-stress markers: CHOP, BiP and phospho-eIF2α in A. hydrophila-infected headkidney macrophages (HKM) in Clarias batrachus. Pre-treatment with ER-stress inhibitor, 4-PBA alleviated ER-stress and HKM apoptosis suggesting ER-UPR critical for the process. The ER-Ca2+ released via inositol-triphosphate and ryanodine receptors induced calpain-2 mediated superoxide ion generation and consequent NF-κB activation. Inhibiting NF-κB activation attenuated NO production suggesting the pro-apoptotic role of NF-κB on HKM pathology. Calpain-2 activated caspase-12 to intensify the apoptotic cascade through mitochondrial-membrane potential (ψm) dissipation and caspase-9 activation. Altered mitochondrial ultra-structure consequent to ER-Ca2+ uptake via uniporters reduced ψm and released cytochrome C. Nitric oxide induced the cGMP/PKG-dependent activation of caspase-8 and truncated-Bid formation. Both the caspases converge onto caspase-3 to execute HKM apoptosis. These findings offer a possible molecular explanation for A. hydrophila pathogenicity. PMID:25059203

  19. Detection of Aeromonas hydrophila in Liquid Media by Volatile Production Similarity Patterns, Using a FF-2A Electronic Nose

    PubMed Central

    Fujioka, Kouki; Arakawa, Eiji; Kita, Jun-ichi; Aoyama, Yoshihiro; Manome, Yoshinobu; Ikeda, Keiichi; Yamamoto, Kenji

    2013-01-01

    A technique for rapid detection of pathogenic microorganisms is essential for the diagnosis of associated infections and for food safety analysis. Aeromonas hydrophila is one such food contaminant. Several methods for rapid detection of this pathogen have been developed; these include multiplex polymerase chain reaction assays and the colony overlay procedure for peptidases. However, these conventional methods can only be used to detect the microorganisms at high accuracy after symptomatic onset of the disease. Therefore, in the future, simple pre-screening methods may be useful for preventing food poisoning and disease. In this paper, we present a novel system for the rapid detection of the microorganism A. hydrophila in cultured media (in <2 h), with the use of an electronic nose (FF-2A). With this electronic nose, we detected the changes of volatile patterns produced by A. hydrophila after 30 min culture. Our calculations revealed that the increased volatiles were similar to the odours of organic acids and esters. In future, distinctive volatile production patterns of microorganisms identified with the electronic nose may have the potential in microorganism detection. PMID:23296330

  20. Plant polysaccharides used as immunostimulants enhance innate immune response and disease resistance against Aeromonas hydrophila infection in fish.

    PubMed

    Wang, Erlong; Chen, Xia; Wang, Kaiyu; Wang, Jun; Chen, Defang; Geng, Yi; Lai, Weimin; Wei, Xianchao

    2016-12-01

    Plant polysaccharides (PPS) are an important medicinal plant product, and play a major role in preventing and controlling infectious microbes in aquaculture. The present study investigated the effect of three PPS; Ficus carica polysaccharides (FCPS), Radix isatidis polysaccharides (RIPS), and Schisandra chinensis polysaccharides (SCPS), used as feed additives, on innate immune responses and disease resistance against Aeromonas hydrophila in crucian carp. Results show that crucian carp fed with these PPS showed significant (p < 0.05) enhancement of their innate immune response including leukocyte phagocytosis activity, serum bactericidal activity, lysozyme activity, total protein level, complement C3, and superoxide dismutase activity compared with the control group. Their degree of influence on these immune parameters was in the order of FCPS > RIPS > SCPS, except for lysozyme activity (RIPS > FCPS > SCPS). In addition, fish cumulative mortalities in the three treatment groups were remarkably lower than in the control group (95%) when challenged with A. hydrophila, relative percent survivals were 57.9%, 47.4%, and 42.1% in FCPS, RIPS, and SCPS groups, respectively. These results suggest that FCPS, RIPS, and SCPS used as immunostimulants are capable of enhancing immune responses and disease resistance against A. hydrophila in crucian carp, and that FCPS was the most effective. The findings from this study will help accelerate research of this topic, and promote the application and development of immunostimulants, such as Chinese herbs, in aquaculture.

  1. Molecular analysis of type II topoisomerases of Aeromonas hydrophila isolated from fish and levofloxacin-induced resistant isolates in vitro.

    PubMed

    Hu, Ruixue; Du, Na; Chen, Nan; Lin, Li; Zhai, Yanhua; Gu, Zemao

    2016-05-01

    The mechanisms of resistance to levofloxacin for Aeromonas hydrophila isolated from diseased fish and selected in vitro were examined in this study. Levofloxacin-resistant mutants were obtained by selection of A. hydrophila in vitro. The quinolone resistance-determining regions (QRDRs) of the gyrA and parC genes were sequenced in Lev(R) strains and reverse mutation strains. All Lev(R) strains carried a point mutation at codon 83 (Ser → Ile), and one strain (25 %) harbored a mutation at position 92 (Leu → Met) in the GyrA-QRDR. After being transferred in a levofloxacin-free medium, one strain of the mutants was successfully reversed and the reversion was related with mutations of GyrA-QRDR at positions 81 (Gly → Asp) and 83 (Ile → Ser). No amino acid alteration was found in the ParC-QRDR. In addition, the minimum inhibitory concentration (MIC) of levofloxacin for the mutants was lower in the presence of reserpine, and all mutants were also resistant to some of the other quinolones. It was found that the mechanism of levofloxacin resistance of A. hydrophila selected in vitro was related to gyrA of type II topoisomerase, and an efflux mechanism was involved in the resistance as well.

  2. Effect of a new recombinant Aeromonas hydrophila vaccine on the grass carp intestinal microbiota and correlations with immunological responses.

    PubMed

    Liu, Lei; Gong, Yu-Xin; Zhu, Bin; Liu, Guang-Lu; Wang, Gao-Xue; Ling, Fei

    2015-07-01

    Intestinal microbiota has become an integral component of the fish, and plays a key role in host metabolism, immunity and health maintenance. However, information on the immune responses after vaccine administration in relation to the intestinal microbiota is absent in fish. The present study focused on the effect of a new recombinant Aeromonas hydrophila vaccine (Aera) by using a novel functionalized, single-walled carbon nanotubes (SWCNTs) as a delivery vehicle on the intestinal microbiota of grass carp (Ctenopharyngodon idella) through the bath immunization, and further explored the immunological responses in intestine, kidney and spleen. By performing deep sequencing, a total of 81,979 valid reads and 609 OTUs obtained from 4 intestine samples were analyzed. We detected 141 genuses, most of which belonged to Firmicutes, Fusobacteria and Proteobacteria. Of note, the quantity of Aeromonas in library Aera (after 6 h Aera vaccine pretreatment, fish were transferred to tanks without SWCNTs-Aera for 28 d) and Aera-GD (6 h Aera vaccine pretreated prior to the group injected by A. hydrophila) was declined 6.5% and 14.6% compared with the control, respectively. Moreover, the expression of seven immune-related genes (IFN-I, TNF-α, CRP, IL-8, IgM, MHC I and CD8α) in the intestine, kidney and spleen of Aera treated fish was significantly enhanced, which indicated that a better tissue immune response in grass carp was induced by the SWCNTs-Aera vaccine. Therefore, a new recombinant SWCNT-Aera vaccine may represent potentially efficient and immunological role in grass carp intestine to resist A. hydrophila infection.

  3. Isolation and Identification of Aeromonas hydrophila and Saprolegnia sp. on Catfish (Clarias gariepinus) in Floating cages in Bozem Moro Krembangan Surabaya

    NASA Astrophysics Data System (ADS)

    Kusdarwati, Rahayu; Kismiyati; Sudarno; Kurniawan, Hendi; Teguh Prayogi, Yudha

    2017-02-01

    Catfish (Clarias gariepinus) is one of the familiar freshwater fish cultured in Indonesia farmer. One of the reason is the high mortality of the catfish infected by Aeromonas hydrophila and Saprolegnia sp. Motile Aeromonas septicemia (MAS) is a common bacterial disease, caused by Aeromonas hydrophila, which affects freshwater fish. In Southeast Asia, the outbreak of this disease was firstly reported from West Java in 1980, when a total of 82.5 tons a month of catfish were lost, while in Central Java in 1984, the total loss was 1.6 tons. Saprolegniosis can cause economic loss due to high mortality from its case reaching 10% to 50%. This research aimed to identify and determine the percentage of A. hydrophila and Saprolegnia sp. the catfish farmed in bozem Moro Krembangan, Surabaya, East Java. Meanwhile, a supporting parameter in this research is the value of water quality parameter including pH, temperature, ammonia and dissolved oxygen that were measured during sampling. The results showed that of the 20 samples taken from the two cages, 19 fish were positively infected by A. drophila. percentage of infections of A. hydrophila that infect umbo catfish in Moro Krembangan, was 95%, while the percentage Saprolegnia sp. was 90%.

  4. Experimental infections of Rana esculenta with Aeromonas hydrophila: a molecular mechanism for the control of the normal flora.

    PubMed

    Simmaco, M; Mangoni, M L; Boman, A; Barra, D; Boman, H G

    1998-10-01

    Frogs can be useful models for studying the mechanisms that may regulate their natural microbial flora. Their skin glands produce a secretion containing 20-30 different peptides, some antimicrobial some neurotrophic. As they often live in soil or silt that is rich in microbes, they can be expected to be able to prevent or eliminate infections in very short periods of time. The bacterium Aeromonas hydrophila is widely distributed in nature and is considered as part of the natural flora of frogs and many animals, including humans. From an alternative frog strain of A. hydrophila, Bo-3, we isolated a spontaneous and stable mutant (Bo-3N), resistant to nalidixic acid, here used to follow the host-microbe interactions in experimental infection of mouth and skin of Rana esculenta. The skin peptides had been previously isolated, sequenced and cloned. We showed that skin treatment with a glucocorticoid (GC) cream blocked de novo synthesis of these peptides and, simultaneously, prepropeptide mRNAs disappeared while IkappaBalpha was up-regulated. Experimental mouth infections with 20 million cells of A. hydrophila Bo-3N showed that a normal wild frog can eliminate the bacteria from the mouth within 15 min, while a frog pretreated with GC cream for 1 h could not reduce Bo-3N below 3500 colony-forming units (CFU)/5 microl 'saliva'. An in vitro comparison showed that frog blood or serum allowed bacteria to grow, while the skin secretion killed the bacteria within 10 min. Using different enzyme-linked immunosorbent assays (ELISAs) with rabbit anti-Bo-3 serum as a positive control, we were able to rule out immunoglobulin G (IgG) binding to A. hydrophila. An assay for immunoglobulin M (IgM) (or some other serum component) in frog serum showed binding to A. hydrophila only corresponding to a few per cent of the positive control. For skin infections we bathed the frogs for 10 min in an overnight culture of Bo-3N diluted to about 10(7) CFU/ml. Electrical stimulation after the bath

  5. Effect of Lactobacillus plantarum C014 on Innate Immune Response and Disease Resistance against Aeromonas hydrophila in Hybrid Catfish

    PubMed Central

    Butprom, Sureerat; Phumkhachorn, Parichat; Rattanachaikunsopon, Pongsak

    2013-01-01

    A bacterial strain isolated from intestines of hybrid catfish (Clarias gariepinus Male × Clarias macrocephalus Female) exhibited an in vitro inhibitory effect on a fish pathogen, Aeromonas hydrophila TISTR 1321. By using the 16S rDNA sequence analysis, it was identified as Lactobacillus plantarum C014. To examine whether L. plantarum C014 had potential for use as an immunostimulant and biocontrol agent in hybrid catfish, the fish diet supplemented with L. plantarum C014 (107 CFU/g diet) was prepared and used for the in vivo investigation of its effect on innate immune response and disease resistance of hybrid catfish. Two innate immune response parameters, phagocytic activity of blood leukocytes and plasma lysozyme activity, were significantly enhanced in the treated fish after 45 days of feeding. Feeding the fish with the L. plantarum C014 supplemented diet for 45 days before challenging them with A. hydrophila at the dose of LD50 could reduce the mortality rate of the fish from 50% (in control group) to 0% (in treated group). Based on its origin and beneficial effect on innate immune response and disease resistance, L. plantarum C014 may be a potential candidate for use as a natural and safe immunostimulant and biocontrol agent in hybrid catfish. PMID:24453842

  6. Development and efficacy of novobiocin and rifampicin-resistant Aeromonas hydrophila as novel vaccines in channel catfish and Nile tilapia.

    PubMed

    Pridgeon, Julia W; Klesius, Phillip H

    2011-10-19

    Three attenuated Aeromonas hydrophila vaccines were developed from the virulent 2009 West Alabama isolates through selection for resistance to both novobiocin and rifampicin. When channel catfish (Ictalurus punctatus) were IP injected with 4×105 colony-forming unit (CFU) of the mutants, no fish died. However, when the same age and size matched channel catfish were IP injected with similar amount of their virulent parents, 80-100% fish died. Similarly, when Nile tilapia (Oreochromis niloticus) were IP injected with 2×108 CFU of the mutants, no fish died. However, when Nile tilapia were IP injected with similar amount of the mutants, all fish died. Vaccination of channel catfish with the mutants at dose of 4×105 CFU/fish offered 86-100% protection against their virulent parents at 14 days post vaccination (dpv). Vaccination of Nile tilapia with the mutants at dose of 2×108 CFU/fish offered 100% protection against their virulent parents at 14, 28, and 56 dpv. Agglutination assay results suggested that protection elicited by the mutants was partially due to antibody-mediated immunity. Taken together, our results suggest that the three attenuated vaccines might be used to protect channel catfish and Nile tilapia against the highly virulent 2009 West Alabama isolates of A. hydrophila.

  7. Influence of clove oil on certain quorum-sensing-regulated functions and biofilm of Pseudomonas aeruginosa and Aeromonas hydrophila.

    PubMed

    Husain, Fohad Mabood; Ahmad, Iqbal; Asif, Mohammad; Tahseen, Qudsia

    2013-12-01

    Quorum sensing (QS) plays an important role in virulence, biofilm formation and survival of many pathogenic bacteria including Pseudomonas aeruginosa. This signalling pathway is considered as novel and promising target for anti-infective agents. In the present investigation, effect of the Sub-MICs of clove oil on QS regulated virulence factors and biofilm formation was evaluated against P. aeruginosa PAO1 and Aeromonas hydrophila WAF-38 strain. Sub-inhibitory concentrations of the clove oil demonstrated statistically significant reduction of las- and rhl-regulated virulence factors such as LasB, total protease, chitinase and pyocyanin production, swimming motility and exopolysaccharide production. The biofilm forming capability of PAO1 and A. hydrophila WAF-38 was also reduced in a concentration-dependent manner at all tested sub-MIC values. Further, the PAO1-preinfected Caenorhabditis elegans displayed an enhanced survival when treated with 1.6 percent v/v of clove oil. The above findings highlight the promising anti-QS-dependent therapeutic function of clove oil against P. aeruginosa.

  8. Polar Glycosylated and Lateral Non-Glycosylated Flagella from Aeromonas hydrophila Strain AH-1 (Serotype O11).

    PubMed

    Fulton, Kelly M; Mendoza-Barberá, Elena; Twine, Susan M; Tomás, Juan M; Merino, Susana

    2015-11-27

    Polar and but not lateral flagellin proteins from Aeromonas hydrophila strain AH-1 (serotype O11) were found to be glycosylated. Top-down mass spectrometry studies of purified polar flagellins suggested the presence of a 403 Da glycan of mass. Bottom-up mass spectrometry studies showed the polar flagellin peptides to be modified with 403 Da glycans in O-linkage. The MS fragmentation pattern of this putative glycan was similar to that of pseudaminic acid derivative. Mutants lacking the biosynthesis of pseudaminic acid (pseB and pseI homologues) were unable to produce polar flagella but no changes were observed in lateral flagella by post-transcriptional regulation of the flagellin. Complementation was achieved by reintroduction of the wild-type pseB and pseI. We compared two pathogenic features (adhesion to eukaryotic cells and biofilm production) between the wild-type strain and two kinds of mutants: mutants lacking polar flagella glycosylation and lacking the O11-antigen lipopolysaccharide (LPS) but with unaltered polar flagella glycosylation. Results suggest that polar flagella glycosylation is extremely important for A. hydrophila AH-1 adhesion to Hep-2 cells and biofilm formation. In addition, we show the importance of the polar flagella glycosylation for immune stimulation of IL-8 production via toll-"like" receptor 5 (TLR5).

  9. Cross-talk among flesh-eating Aeromonas hydrophila strains in mixed infection leading to necrotizing fasciitis.

    PubMed

    Ponnusamy, Duraisamy; Kozlova, Elena V; Sha, Jian; Erova, Tatiana E; Azar, Sasha R; Fitts, Eric C; Kirtley, Michelle L; Tiner, Bethany L; Andersson, Jourdan A; Grim, Christopher J; Isom, Richard P; Hasan, Nur A; Colwell, Rita R; Chopra, Ashok K

    2016-01-19

    Necrotizing fasciitis (NF) caused by flesh-eating bacteria is associated with high case fatality. In an earlier study, we reported infection of an immunocompetent individual with multiple strains of Aeromonas hydrophila (NF1-NF4), the latter three constituted a clonal group whereas NF1 was phylogenetically distinct. To understand the complex interactions of these strains in NF pathophysiology, a mouse model was used, whereby either single or mixed A. hydrophila strains were injected intramuscularly. NF2, which harbors exotoxin A (exoA) gene, was highly virulent when injected alone, but its virulence was attenuated in the presence of NF1 (exoA-minus). NF1 alone, although not lethal to animals, became highly virulent when combined with NF2, its virulence augmented by cis-exoA expression when injected alone in mice. Based on metagenomics and microbiological analyses, it was found that, in mixed infection, NF1 selectively disseminated to mouse peripheral organs, whereas the other strains (NF2, NF3, and NF4) were confined to the injection site and eventually cleared. In vitro studies showed NF2 to be more effectively phagocytized and killed by macrophages than NF1. NF1 inhibited growth of NF2 on solid media, but ExoA of NF2 augmented virulence of NF1 and the presence of NF1 facilitated clearance of NF2 from animals either by enhanced priming of host immune system or direct killing via a contact-dependent mechanism.

  10. Identification and Characterization of an Aeromonas hydrophila Oligopeptidase Gene pepF Negatively Related to Biofilm Formation

    PubMed Central

    Du, Hechao; Pang, Maoda; Dong, Yuhao; Wu, Yafeng; Wang, Nannan; Liu, Jin; Awan, Furqan; Lu, Chengping; Liu, Yongjie

    2016-01-01

    Bacterial biofilms are involved in adaptation to complex environments and are responsible for persistent bacterial infections. Biofilm formation is a highly complex process during which multifarious genes work together regularly. In this study, we screened the EZ-Tn5 transposon mutant library to identify genes involved in biofilm formation of Aeromonas hydrophila. A total of 24 biofilm-associated genes were identified, the majority of which encoded proteins related to cell structure, transcription and translation, gene regulation, growth and metabolism. The mutant strain TM90, in which a gene encoding oligopeptidase F (pepF) was disturbed, showed significant upregulation of biofilm formation compared to the parental strain. The TM90 colony phenotype was smaller, more transparent, and splendent. The adhesive ability of TM90 to HEp-2 cells was significantly increased compared with the parental strain. Fifty percent lethal dose (LD50) determinations in zebrafish demonstrated that the enhanced-biofilm mutant TM90 was highly attenuated relative to the wild-type strain. In conclusion, the pepF gene is demonstrated for the first time to be a negative factor for biofilm formation and is involved in A. hydrophila pathogenicity. PMID:27713736

  11. Oreochromis mossambicus diet supplementation with Psidium guajava leaf extracts enhance growth, immune, antioxidant response and resistance to Aeromonas hydrophila.

    PubMed

    Gobi, Narayanan; Ramya, Chinnu; Vaseeharan, Baskaralingam; Malaikozhundan, Balasubramanian; Vijayakumar, Sekar; Murugan, Kadarkarai; Benelli, Giovanni

    2016-11-01

    In this research, we focused on the efficacy of aqueous and ethanol leaf extracts of Psidium guajava L. (guava) based experimental diets on the growth, immune, antioxidant and disease resistance of tilapia, Oreochromis mossambicus following challenge with Aeromonas hydrophila. The experimental diets were prepared by mixing powdered (1, 5 and 10 mg/g) aqueous and ethanol extract of guava leaf with commercial diet. The growth (FW, FCR and SGR), non-specific cellular immune (myeloperoxidase activity, reactive oxygen activity and reactive nitrogen activity) humoral immune (complement activity, antiprotease, alkaline phosphatase activity and lysozyme activity) and antioxidant enzyme responses (SOD, GPX, and CAT) were examined after 30 days of post-feeding. A significant enhancement in the biochemical and immunological parameters of fish were observed fed with experimental diets compared to control. The dietary supplementation of P. guajava leaf extract powder for 30 days significantly reduced the mortality and increased the disease resistance of O. mossambicus following challenge with A. hydrophila at 50 μl (1 × 10(7) cells ml(-1)) compared to control after post-infection. The results suggest that the guava leaf extract could be used as a promising feed additive in aquaculture.

  12. Role of Hcp, a type 6 secretion system effector, of Aeromonas hydrophila in modulating activation of host immune cells.

    PubMed

    Suarez, Giovanni; Sierra, Johanna C; Kirtley, Michelle L; Chopra, Ashok K

    2010-12-01

    Recently, we reported that the type 6 secretion system (T6SS) of Aeromonas hydrophila SSU plays an important role in bacterial virulence in a mouse model, and immunization of animals with the T6SS effector haemolysin co-regulated protein (Hcp) protected them against lethal infections with wild-type bacteria. Additionally, we showed that the mutant bacteria deleted for the vasH gene within the T6SS gene cluster did not express the hcp gene, while the vasK mutant could express and translocate Hcp, but was unable to secrete it into the extracellular milieu. Both of these A. hydrophila SSU mutants were readily phagocytosed by murine macrophages, pointing to the possible role of the secreted form of Hcp in the evasion of the host innate immunity. By using the ΔvasH mutant of A. hydrophila, our in vitro data showed that the addition of exogenous recombinant Hcp (rHcp) reduced bacterial uptake by macrophages. These results were substantiated by increased bacterial virulence when rHcp was added along with the ΔvasH mutant in a septicaemic mouse model of infection. Analysis of the cytokine profiling in the intraperitoneal lavage as well as activation of host cells after 4 h of infection with the ΔvasH mutant supplemented with rHcp indicated that this T6SS effector inhibited production of pro-inflammatory cytokines and induced immunosuppressive cytokines, such as interleukin-10 and transforming growth factor-β, which could circumvent macrophage activation and maturation. This mechanism of innate immune evasion by Hcp possibly inhibited the recruitment of cellular immune components, which allowed bacterial multiplication and dissemination in animals, thereby leading to their mortality.

  13. Temporal and spatial fate of GFP-expressing motile and nonmotile Aeromonas hydrophila in the house fly digestive tract.

    PubMed

    McGaughey, Joseph; Nayduch, Dana

    2009-01-01

    To gain insight into the transmissibility of bacteria by house flies, the temporal and spatial fate of green fluorescent protein (GFP)-expressing motile and nonmotile strains of Aeromonas hydrophila was examined within the alimentary canal. Liquid food consumed by house flies is first stored in the crop and then is regurgitated and/or passed into the midgut. Once within the midgut, food is contained inside a double-layered peritrophic matrix (PM), with the inner layer enveloping digested material and forming fecal pellets for excretion. Between 1 and 12 h after ingestion, and irrespective of motility, live GFP+ A. hydrophila adhered to the luminal surfaces of the crop and inner PM of bacteria-fed flies. However, some nonadherent, motile bacteria moved freely within the PM lumen in the anteromedial midgut, whereas fecal pellets (lysed bacteria) continued passing posteriorly. At 12-24 h, adhered bacteria of both strains were lysed in the distal midgut, compressed into fecal pellets, and excreted. Viable bacteria in the crop visually exceeded numbers within these fecal pellets. Culture recovery at the same time points showed a 1,000-fold increase of viable bacteria at 2 h, presumably in the crop, with a temporal decline thereafter. Further, viable bacteria were recovered from vomit specks and orally contaminated substrates up to 2 h after feeding but never from feces. These results suggest that A. hydrophila is a transient resident of the house fly alimentary canal and is only orally transmissible for a short time after ingestion. Thus, regurgitation may be more significant than fecal transmission in the spread of some house fly transmitted bacterial diseases.

  14. Transcriptome Analysis of the Innate Immunity-Related Complement System in Spleen Tissue of Ctenopharyngodon idella Infected with Aeromonas hydrophila

    PubMed Central

    Dang, Yunfei; Xu, Xiaoyan; Shen, Yubang; Hu, Moyan; Zhang, Meng; Li, Lisen; Lv, Liqun; Li, Jiale

    2016-01-01

    The grass carp (Ctenopharyngodon idella) is an important commercial farmed herbivorous fish species in China, but is susceptible to Aeromonas hydrophila infections. In the present study, we performed de novo RNA-Seq sequencing of spleen tissue from specimens of a disease-resistant family, which were given intra-peritoneal injections containing PBS with or without a dose of A. hydrophila. The fish were sampled from the control group at 0 h, and from the experimental group at 4, 8, 12, 24, 48 and 72 h. 122.18 million clean reads were obtained from the normalized cDNA libraries; these were assembled into 425,260 contigs and then 191,795 transcripts. Of those, 52,668 transcripts were annotated with the NCBI Nr database, and 41,347 of the annotated transcripts were assigned into 90 functional groups. 20,569 unigenes were classified into six main categories, including 38 secondary KEGG pathways. 2,992 unigenes were used in the analysis of differentially expressed genes (DEGs). 89 of the putative DEGs were related to the immune system and 41 of them were involved in the complement and coagulation cascades pathway. This study provides insights into the complement and complement-related pathways involved in innate immunity, through expression profile analysis of the genomic resources in C. idella. We conclude that complement and complement-related genes play important roles during defense against A. hydrophila infection. The immune response is activated at 4 h after the bacterial injections, indicating that the complement pathways are activated at the early stage of bacterial infection. The study has improved our understanding of the immune response mechanisms in C. idella to bacterial pathogens. PMID:27383749

  15. Molecular characterization of an interleukin-4/13B homolog in grass carp (Ctenopharyngodon idella) and its role in fish against Aeromonas hydrophila infection.

    PubMed

    Yang, Zhi-Jing; Li, Chang-Hong; Chen, Jie; Zhang, Hao; Li, Ming-Yun; Chen, Jiong

    2016-10-01

    Mammalian interleukin 4 (IL-4) and interleukin 13 (IL-13) molecules are anti-inflammatory cytokines mediating the alternative activation of macrophages. However, the role of fish IL-4/13 homologs in monocytes/macrophages (MO/MФ) polarization remains unclear. In this study, we have functionally identified an IL-4/13B homolog in grass carp (Ctenopharyngodon idella), which is termed as CiIL-4/13B. Multiple alignment showed that CiIL-4/13B shared the typical characteristics and structure with other known fish IL-4/13. Phylogenetic analysis showed that CiIL-4/13B is evolutionarily closely related to zebrafish (Danio rerio) and common carp (Cyprinus carpio) IL-4/13B. CiIL-4/13B mRNA was constitutively expressed in tissues and peripheral blood lymphocytes (PBLs) examined, with its highest expression seen in PBLs. Following Aeromonas hydrophila infection, CiIL-4/13B mRNA expression was upregulated. Recombinant CiIL-4/13B (rCiIL-4/13B) was overexpressed in Escherichia coli and purified for a functional study. Using prepared anti-rCiIL-4/13B antiserum, Western blot analysis showed that native CiIL-4/13B in grass carp plasma is N-glycosylated. Intraperitoneal injection of bioactive rCiIL-4/13B significantly increased the survival rate of grass carp against A. hydrophila, and decreased the tissue bacterial load, with a higher dose having better effects. Bioactive rCiIL-4/13B treatment decreased nitrite production and mRNA expression of proinflammatory cytokines (IL-1β and TNF-α), while it increased arginase activity and mRNA expression of anti-inflammatory cytokines (TGF-β and IL-10). The phagocytosis by grass carp MO/MФ had no significant changes by the 8 h treatment of bioactive rCiIL-4/13B compared to that of the negative control, while it was significantly inhibited by the 24 h treatment of bioactive rCiIL-4/13B. The inhibitory effect of rCiIL-4/13B on MO/MФ phagocytosis may be a consequence of MO/MФ proliferation. In summary, our results suggest that CiIL-4/13B

  16. Determination of the Viability of Aeromonas hydrophila in Different Types of Water by Flow Cytometry, and Comparison with Classical Methods

    PubMed Central

    Pianetti, Anna; Falcioni, Tania; Bruscolini, Francesca; Sabatini, Luigia; Sisti, Elivio; Papa, Stefano

    2005-01-01

    The presence of Aeromonas spp. in water can represent a risk for human health. Therefore, it is important to know the physiological status of these bacteria and their survival in the environment. We studied the behavior of a strain of Aeromonas hydrophila in river water, spring water, brackish water, mineral water, and chlorinated drinking water, which had different physical and chemical characteristics. The bacterial content was evaluated by spectrophotometric and plate count techniques. Flow cytometric determination of viability was carried out using a dual-staining technique that enabled us to distinguish viable bacteria from damaged and membrane-compromised bacteria. The traditional methods showed that the bacterial content was variable and dependent on the type of water. The results obtained from the plate count analysis correlated with the absorbance data. In contrast, the flow cytometric analysis results did not correlate with the results obtained by traditional methods; in fact, this technique showed that there were viable cells even when the optical density was low or no longer detectable and there was no plate count value. According to our results, flow cytometry is a suitable method for assessing the viability of bacteria in water samples. Furthermore, it permits fast detection of bacteria that are in a viable but nonculturable state, which are not detectable by conventional methods. PMID:16332772

  17. Activation of the Complement Classical Pathway (C1q Binding) by Mesophilic Aeromonas hydrophila Outer Membrane Protein

    PubMed Central

    Merino, Susana; Nogueras, Maria Mercedes; Aguilar, Alicia; Rubires, Xavier; Albertí, Sebastian; Benedí, Vicente Javier; Tomás, Juan M.

    1998-01-01

    The mechanism of killing of Aeromonas hydrophila serum-sensitive strains in nonimmune serum by the complement classical pathway has been studied. The bacterial cell surface component that binds C1q more efficiently was identified as a major outer membrane protein of 39 kDa, presumably the porin II described by D. Jeanteur, N. Gletsu, F. Pattus, and J. T. Buckley (Mol. Microbiol. 6:3355–3363, 1992), of these microorganisms. We have demonstrated that the purified form of porin II binds C1q and activates the classical pathway in an antibody-independent manner, with the subsequent consumption of C4 and reduction of the serum total hemolytic activity. Activation of the classical pathway has been observed in human nonimmune serum and agammaglobulinemic serum (both depleted of factor D). Binding of C1q to other components of the bacterial outer membrane, in particular to rough lipopolysaccharide, could not be demonstrated. Activation of the classical pathway by this lipopolysaccharide was also much less efficient than activation by the outer membrane protein. The strains possessing O-antigen lipopolysaccharide bind less C1q than the serum-sensitive strains, because the outer membrane protein is less accessible, and are resistant to complement-mediated killing. Finally, a similar or identical outer membrane protein (presumably porin II) that binds C1q was shown to be present in strains from the most common mesophilic Aeromonas O serogroups. PMID:9673268

  18. Thin-film fixed-bed reactor (TFFBR) for solar photocatalytic inactivation of aquaculture pathogen Aeromonas hydrophila

    PubMed Central

    2012-01-01

    Background Outbreaks of infectious diseases by microbial pathogens can cause substantial losses of stock in aquaculture systems. There are several ways to eliminate these pathogens including the use of antibiotics, biocides and conventional disinfectants, but these leave undesirable chemical residues. Conversely, using sunlight for disinfection has the advantage of leaving no chemical residue and is particularly suited to countries with sunny climates. Titanium dioxide (TiO2) is a photocatalyst that increases the effectiveness of solar disinfection. In recent years, several different types of solar photocatalytic reactors coated with TiO2 have been developed for waste water and drinking water treatment. In this study a thin-film fixed-bed reactor (TFFBR), designed as a sloping flat plate reactor coated with P25 DEGUSSA TiO2, was used. Results The level of inactivation of the aquaculture pathogen Aeromonas hydrophila ATCC 35654 was determined after travelling across the TFFBR under various natural sunlight conditions (300-1200 W m-2), at 3 different flow rates (4.8, 8.4 and 16.8 L h-1). Bacterial numbers were determined by conventional plate counting using selective agar media, cultured (i) under conventional aerobic conditions to detect healthy cells and (ii) under conditions designed to neutralise reactive oxygen species (agar medium supplemented with the peroxide scavenger sodium pyruvate at 0.05% w/v, incubated under anaerobic conditions), to detect both healthy and sub-lethally injured (oxygen-sensitive) cells. The results clearly demonstrate that high sunlight intensities (≥ 600 W m-2) and low flow rates (4.8 L h-1) provided optimum conditions for inactivation of A. hydrophila ATCC 3564, with greater overall inactivation and fewer sub-lethally injured cells than at low sunlight intensities or high flow rates. Low sunlight intensities resulted in reduced overall inactivation and greater sub-lethal injury at all flow rates. Conclusions This is the first

  19. Transcriptome analysis and microsatellite discovery in the blunt snout bream (Megalobrama amblycephala) after challenge with Aeromonas hydrophila.

    PubMed

    Tran, Ngoc Tuan; Gao, Ze-Xia; Zhao, Hong-Hao; Yi, Shao-Kui; Chen, Bo-Xiang; Zhao, Yu-Hua; Lin, Li; Liu, Xue-Qin; Wang, Wei-Min

    2015-07-01

    The blunt snout bream, Megalobrama amblycephala, is a herbivorous freshwater fish species native to China and a major aquaculture species in Chinese freshwater polyculture systems. In recent years, the bacterium Aeromonas hydrophila has been reported to be its pathogen causing great losses of farmed fish. To understand the immune response of the blunt snout bream to A. hydrophila infection, we used the Solexa/Illumina technology to analyze the transcriptomic profile after artificial bacterial infection. Two nonnormalized cDNA libraries were synthesized from tissues collected from control blunt snout bream or those injected with A. hydrophila. After assembly, 155,052 unigenes (average length 692.8 bp) were isolated. All unigenes were annotated using BLASTX relative to several public databases: the National Center for Biotechnology Information nonreduntant (Nr) database, SwissProt, Eukaryotic Orthologous Groups of proteins (KOG), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO). The sequence similarity (86%) of the assembled unigenes was to zebrafish based on the Nr database. A number of unigenes (n = 30,482) were assigned to three GO categories: biological processes (25,242 unigenes), molecular functions (26,096 unigenes), and cellular components (22,778 unigenes). 20,909 unigenes were classified into 25 KOG categories and 28,744 unigenes were assigned into 315 specific signaling pathways. In total, 238 significantly differentially expressed unigenes (mapped to 125 genes) were identified: 101 upregulated genes and 24 downregulated genes. Another 303 unigenes were mapped to unknown or novel genes. Among the known expressed genes identified, 53 were immune-related genes and were distributed in 71 signaling pathways. The expression patterns of selected up- and downregulated genes from the control and injected groups were determined with reverse transcription-quantitative PCR (RT-qPCR). Microsatellites (n = 10,877), including di-to pentanucleotide

  20. A linkage map of transcribed single nucleotide polymorphisms in rohu (Labeo rohita) and QTL associated with resistance to Aeromonas hydrophila

    PubMed Central

    2014-01-01

    Background Production of carp dominates world aquaculture. More than 1.1 million tonnes of rohu carp, Labeo rohita (Hamilton), were produced in 2010. Aeromonas hydrophila is a bacterial pathogen causing aeromoniasis in rohu, and is a major problem for carp production worldwide. There is a need to better understand the genetic mechanisms affecting resistance to this disease, and to develop tools that can be used with selective breeding to improve resistance. Here we use a 6 K SNP array to genotype 21 full-sibling families of L. rohita that were experimentally challenged intra-peritoneally with a virulent strain of A. hydrophila to scan the genome for quantitative trait loci associated with disease resistance. Results In all, 3193 SNPs were found to be informative and were used to create a linkage map and to scan for QTL affecting resistance to A. hydrophila. The linkage map consisted of 25 linkage groups, corresponding to the number of haploid chromosomes in L. rohita. Male and female linkage maps were similar in terms of order, coverage (1384 and 1393 cM, respectively) and average interval distances (1.32 and 1.35 cM, respectively). Forty-one percent of the SNPs were annotated with gene identity using BLAST (cut off E-score of 0.001). Twenty-one SNPs mapping to ten linkage groups showed significant associations with the traits hours of survival and dead or alive (P <0.05 after Bonferroni correction). Of the SNPs showing significant or suggestive associations with the traits, several were homologous to genes of known immune function or were in close linkage to such genes. Genes of interest included heat shock proteins (70, 60, 105 and “small heat shock proteins”), mucin (5b precursor and 2), lectin (receptor and CD22), tributyltin-binding protein, major histocompatibility loci (I and II), complement protein component c7-1, perforin 1, ubiquitin (ligase, factor e4b isoform 2 and conjugation enzyme e2 c), proteasome subunit, T-cell antigen receptor and

  1. Aeromonas hydrophila OmpW PLGA Nanoparticle Oral Vaccine Shows a Dose-Dependent Protective Immunity in Rohu (Labeo rohita).

    PubMed

    Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Paul, Joydeb; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Munang'andu, Hetron Mweemba

    2016-06-01

    Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane protein W (OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly d,l-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370-375 nm, encapsulation efficiency of 53% and -19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34% within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to

  2. Aeromonas hydrophila OmpW PLGA Nanoparticle Oral Vaccine Shows a Dose-Dependent Protective Immunity in Rohu (Labeo rohita)

    PubMed Central

    Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Paul, Joydeb; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Munang’andu, Hetron Mweemba

    2016-01-01

    Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane protein W (OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly d,l-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370–375 nm, encapsulation efficiency of 53% and −19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34% within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to

  3. Multifocal Aeromonas Osteomyelitis in a Child with Leukemia

    PubMed Central

    Doganis, Dimitrios; Baka, Margarita; Tsolia, Maria; Pourtsidis, Apostolos; Lebessi, Evangelia; Varvoutsi, Maria; Bouhoutsou, Despina; Kosmidis, Helen

    2016-01-01

    Aeromonas hydrophila is a Gram negative organism causing both intestinal and extraintestinal disease. The case of a 14-year-old girl with underlying immunodeficiency and leukemia who developed systemic A. hydrophila infection is described in this report. While in deep bone marrow aplasia she developed fever, severe pain in the lower extremities, and swelling of the left femur. Blood culture showed Escherichia coli and A. hydrophila whereas pus culture from the soft tissue swelling showed the presence of A. hydrophila. Imaging studies showed diffuse osteolytic lesions. Patient received 5 months of intravenous and oral antibiotics and she improved clinically whereas the radiology findings persisted. PMID:27200197

  4. POTENTIAL FOR GREAT EGRETS (ARDEA ALBA) TO TRANSMIT A VIRULENT STRAIN OF AEROMONAS HYDROPHILA AMONG CHANNEL CATFISH (ICTALURUS PUNCTATUS) CULTURE PONDS.

    PubMed

    Jubirt, Madison M; Hanson, Larry A; Hanson-Dorr, Katie C; Ford, Lorelei; Lemmons, Scott; Fioranelli, Paul; Cunningham, Fred L

    2015-07-01

    Aeromonas hydrophila is a gram-negative, rod-shaped, facultative, anaerobic bacterium that is ubiquitous in freshwater and slightly brackish aquatic environments and infects fish, humans, reptiles, and birds. Recent severe outbreaks of disease in commercial channel catfish (Ictalurus punctatus) aquaculture ponds have been associated with a highly virulent A. hydrophila strain (VAH), which is genetically distinct from less-virulent strains. The epidemiology of this disease has not been determined. Given that A. hydrophila infects birds, we hypothesized that fish-eating birds may serve as a reservoir for VAH and spread the pathogen by flying to uninfected ponds. Great Egrets (Ardea alba) were used in this transmission model because these wading birds frequently prey on farmed catfish. Great Egrets that were fed VAH-infected catfish shed VAH in feces demonstrating their potential to spread VAH.

  5. Quorum sensing by N-acylhomoserine lactones is not required for Aeromonas hydrophila during growth with organic particles in lake water microcosms.

    PubMed

    Styp von Rekowski, Katharina; Hempel, Melanie; Philipp, Bodo

    2008-05-01

    It was investigated whether quorum sensing (QS) mediated by N-acylhomoserine lactones (AHLs) was important for heterotrophic bacteria from the littoral zone of the oligotrophic Lake Constance for growth with organic particles. More than 900 colonies from lake water microcosms with artificial organic aggregates consisting of autoclaved unicellular algae embedded in agarose beads were screened for AHL-production. AHL-producing bacteria of the genus Aeromonas enriched in the microcosms but AHLs could not be detected in any microcosm. To test for a potential function of AHL-mediated QS, growth experiments with the wild type and an AHL-deficient mutant of Aeromonas hydrophila in lake water microcosms were performed. Growth of both strains did not differ in single cultures and showed no mutual influence in co-cultures. In co-cultures with a competitor bacterium belonging to the Cytophaga-Flavobacterium group, growth of both A. hydrophila strains was reduced while growth of the competitor bacterium was not affected. Exogenous AHL-addition did not influence growth of the Aeromonas strains in any microcosm experiment. These results showed that AHL-mediated QS was not required for A. hydrophila during colonization and degradation of organic particles in lake water microcosms, suggesting that cell-cell signalling of heterotrophic bacteria in oligotrophic waters relies on novel signal molecules.

  6. Expression and activity of recombinant proaerolysin derived from Aeromonas hydrophila cultured from diseased channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proaerolysin-coding gene was cloned from the genomic DNA of A. hydrophila and heterologously expressed in E. coli. The purified recombinant proaerolysin was inactive and could be activated by treatment with proteases, furin and trypsin, and extra-cellular proteins (ECPs, the cell-free supernatant of...

  7. Isolation and Antimicrobial Testing of Aeromonas spp., Citrobacter spp., Cronobacter spp., Enterobacter spp., Escherichia spp., Klebsiella spp., and Trabulsiella spp. from the Gallbladder of Pigs.

    PubMed

    Evangelopoulou, Grammato; Filioussis, Georgios; Kritas, Spyridon; Kantere, Maria; Burriel, Angeliki R

    2015-01-01

    The presence of Gram-negative bacteria species, other than Salmonella spp., in the gallbladder of pigs was examined. Isolated Gram-negative bacteria were assigned to species using the Microgen™ GnA+B-ID Systems. Of the 64 isolated strains 43 were identified as Escherichia coli, seven as Enterobacter spp., three each as Klebsiella spp., Citrobacterfreundii, Aeromonas hydrophila and Cronobacter sakazakii and one each as Escherichiafergusonii and Trabulsiella guamensis. Their antibiograms showed very high resistance to ampicillin, amoxicillin, tetracycline, chloramphenicol and sulfamethoxazole/trimethoprim. It was concluded that the pigs' gallbladder is a reservoir of potentially pathogenic Gram-negative bacteria for pork consumers.

  8. The Cytotoxic Enterotoxin of Aeromonas hydrophila Induces Proinflammatory Cytokine Production and Activates Arachidonic Acid Metabolism in Macrophages

    PubMed Central

    Chopra, A. K.; Xu, X.-J.; Ribardo, D.; Gonzalez, M.; Kuhl, K.; Peterson, J. W.; Houston, C. W.

    2000-01-01

    An aerolysin-related cytotoxic enterotoxin (Act) of Aeromonas hydrophila possesses multiple biological activities, which include its ability to lyse red blood cells, destroy tissue culture cell lines, evoke a fluid secretory response in ligated intestinal loop models, and induce lethality in mice. The role of Act in the virulence of the organism has been demonstrated. In this study, we evaluated the potential of Act to induce production of proinflammatory cytokines associated with Act-induced tissue injury and Act's capacity to activate in macrophages arachidonic acid (AA) metabolism that leads to production of eicosanoids (e.g., prostaglandin E2 [PGE2]). Our data indicated that Act stimulated the production of tumor necrosis factor alpha and upregulated the expression of genes encoding interleukin-1β (IL-1β) and IL-6 in the murine macrophage cell line RAW264.7. Act also activated transcription of the gene encoding inducible nitric oxide synthase. Act evoked the production of PGE2 coupled to the cyclooxygenase-2 (COX-2) pathway. AA is a substrate for PGE2, and Act produced AA from phospholipids by inducing group V secretory phospholipase A2. We also demonstrated that Act increased cyclic AMP (cAMP) production in macrophages. cAMP, along with PGE2, could potentiate fluid secretion in animal models because of infiltration and activation of macrophages resulting from Act-induced tissue injury. After Act treatment of RAW cells, we detected an increased translocation of NF-κB and cAMP-responsive element binding protein (CREB) to the nucleus using gel shift assays. Act also upregulated production of antiapoptotic protein Bcl-2 in macrophages, suggesting a protective role for Bcl-2 against cell death induced by proinflammatory cytokines. The increased expression of genes encoding the proinflammatory cytokines, COX-2, and Bcl-2 appeared correlated with the activation of NF-κB and CREB. This is the first report of the detailed mechanisms of action of Act from A

  9. A novel effect of imidazole derivative KK-42 on increasing survival of Aeromonas hydrophila challenged prawn Macrobrachium nipponense.

    PubMed

    Wang, Wen-Feng; Yang, Hong; Liu, Fang; Chen, Xiang-Li; Lv, Yan-Jie; Ning, Qian-Ji

    2013-01-01

    Imidazole derivative KK-42 is well known as the insect growth regulator. Here we find that KK-42 pretreatment could promote the survival of Macrobrachium nipponense infected with Aeromonas hydrophila, which is considered to be possibly related to the prophenoloxidase (proPO), a conserved copper-containing enzyme that plays an important role in defense against pathogens. In this study, a full-length of proPO gene from M. nipponense haemocytes, designated as MnproPO, was firstly cloned and characterized. The full-length cDNA contained 2428 bp with a 2013 bp open reading frame encoding a putative proPO protein of 671 amino acids with a predicted molecular mass of 76.5 kDa and pI of 7.31. It was predicted to possess all the expected features of proPO members, including two putative copper-binding sites with six histidine residues and a thiol ester-like motif. Sequence analysis showed that MnproPO exhibited the highest amino acid sequence similarity (93%) to a proPO of Macrobrachium rosenbergii. The gene was expressed highly in haemocytes and weakly in hepatopancreas. Real-time PCR analysis revealed that the MnproPO expression increased significantly at 3, 12 and 24 h after KK-42 treatment, the PO activity also importantly rose from 6 to 48 h in KK-42-treated prawns and reached the maximum at 24 h with a 2.3-fold higher than that in control group. Injection of A. hydrophila could stimulate the MnproPO transcription and PO activity whether or not the prawns were pretreated by KK-42, the mRNA level increased obviously only at 3 h and 6 h after the bacterium injection (challenged control), but increased constantly during the phase of experiment except at 6 h under the condition of KK-42 pretreatment (challenged treatment group). The change trend of PO activity was basically similar to that of MnproPO expression. Our present results demonstrate that the MnproPO expression as well as PO activity may be induced by KK-42, which is likely one of the molecular mechanisms of KK

  10. Single-walled carbon nanotubes as delivery vehicles enhance the immunoprotective effects of a recombinant vaccine against Aeromonas hydrophila.

    PubMed

    Gong, Yu-Xin; Zhu, Bin; Liu, Guang-Lu; Liu, Lei; Ling, Fei; Wang, Gao-Xue; Xu, Xin-Gang

    2015-01-01

    To reduce the economic losses caused by diseases in aquaculture industry, more efficient and economic prophylactic measures should be urgently investigated. In this research, the effects of a novel functionalized single-walled carbon nanotubes (SWCNTs) applied as a delivery vehicle for recombinant Aeromonas hydrophila vaccine administration via bath or injection in juvenile grass carp were studied. The results showed that SWCNT as a vector for the recombinant protein aerA, augmented the production of specific antibodies, apparently stimulated the induction of immune-related genes, and induced higher level of survival rate compared with free aerA subunit vaccine. Furthermore, we compared the routes of bath and intramuscular injection immunization by SWCNTs-aerA vaccine, and found that similar antibody levels induced by SWCNTs-aerA were observed in both immunization routes. Meanwhile, a similar relative percentage survival (approximately 80%) was found in both a 40 mg/L bath immunization group, and a 20 μg injection group. The results indicate that functionalized SWCNTs could be a promising delivery vehicle to potentiate the immune response of recombinant vaccines, and might be used to vaccinate juvenile fish by bath administration method.

  11. Analysis and Validation of a Predictive Model for Growth and Death of Aeromonas hydrophila under Modified Atmospheres at Refrigeration Temperatures

    PubMed Central

    Pin, Carmen; Velasco de Diego, Raquel; George, Susan; García de Fernando, Gonzalo D.; Baranyi, József

    2004-01-01

    Specific growth and death rates of Aeromonas hydrophila were measured in laboratory media under various combinations of temperature, pH, and percent CO2 and O2 in the atmosphere. Predictive models were developed from the data and validated by means of observations obtained from (i) seafood experiments set up for this purpose and (ii) the ComBase database (http://www.combase.cc; http://wyndmoor.arserrc.gov/combase/).Two main reasons were identified for the differences between the predicted and observed growth in food: they were the variability of the growth rates in food and the bias of the model predictions when applied to food environments. A statistical method is presented to quantitatively analyze these differences. The method was also used to extend the interpolation region of the model. In this extension, the concept of generalized Z values (C. Pin, G. García de Fernando, J. A. Ordóñez, and J. Baranyi, Food Microbiol. 18:539-545, 2001) played an important role. The extension depended partly on the density of the model-generating observations and partly on the accuracy of extrapolated predictions close to the boundary of the interpolation region. The boundary of the growth region of the organism was also estimated by means of experimental results for growth and death rates. PMID:15240265

  12. Knockout of the alanine racemase gene in Aeromonas hydrophila HBNUAh01 results in cell wall damage and enhanced membrane permeability.

    PubMed

    Liu, Dong; Zhang, Lu; Xue, Wen; Wang, Yaping; Ju, Jiansong; Zhao, Baohua

    2015-07-01

    This study focused on the alanine racemase gene (alr-2), which is involved in the synthesis of d-alanine that forms the backbone of the cell wall. A stable alr-2 knockout mutant of Aeromonas hydrophila HBNUAh01 was constructed. When the mutant was supplemented with d-alanine, growth was unaffected; deprivation of d-alanine caused the growth arrest of the starved mutant cells, but not cell lysis. No alanine racemase activity was detected in the culture of the mutant. Additionally, a membrane permeability assay showed increasing damage to the cell wall during d-alanine starvation. No such damage was observed in the wild type during culture. Scanning and transmission electron microscopy analyses revealed deficiencies of the cell envelope and perforation of the cell wall. Leakage of UV-absorbing substances from the mutants was also observed. Thus, the partial viability of the mutants and their independence of d-alanine for growth indicated that inactivation of alr-2 does not impose an auxotrophic requirement for d-alanine.

  13. Transcriptome analysis of the endangered Chinese giant salamander (Andrias davidianus): Immune modulation in response to Aeromonas hydrophila infection.

    PubMed

    Qi, Zhitao; Zhang, Qihuan; Wang, Zisheng; Ma, Tianyi; Zhou, Jie; Holland, Jason W; Gao, Qian

    2016-01-01

    The endangered Chinese giant salamander (Andrias davidianus) is the largest extant amphibian species. Disease outbreaks represent one of the major factors threatening A. davidianus populations in the wild and the viability of artificial breeding programmes. Development of future immune therapies to eliminate infectious disease in A. davidianus is dependent on a thorough understanding of the immune mechanisms elicited by pathogen encounters. To this end we have undertaken, for the first time in amphibians, differential transcriptome analysis of the giant salamander response to Aeromonas hydrophila, one of the most devastating pathogens affecting amphibian populations. Out of 87,204 non-redundant consensus unigenes 19,216 were annotated, 6834 of which were upregulated and 906 down-regulated following bacterial infection. 2058 unigenes were involved with immune system processes, including 287 differentially expressed unigenes indicative of the impact of bacterial infection on several innate and adaptive immune pathways in the giant salamander. Other pathways not directly associated with immune-related activity were differentially expressed, including developmental, structural, molecular and growth processes. Overall, this work provides valuable insights into the underlying immune mechanisms elicited during bacterial infection in amphibians that may aid in the future development of disease control measures in protecting the Chinese giant salamander. With the unique position of amphibians in the transition of tetrapods from aquatic to terrestrial habitats, our study will also be invaluable towards the further understanding of the evolution of tetrapod immunity.

  14. Chlorophytum borivilianum Polysaccharide Fraction Provokes the Immune Function and Disease Resistance of Labeo rohita against Aeromonas hydrophila

    PubMed Central

    Giri, Sib Sankar; Sen, Shib Sankar; Chi, Cheng; Kim, Hyoun Joong; Yun, Saekil; Park, Se Chang; Sukumaran, V.

    2015-01-01

    The present study aimed to investigate the effects of Chlorophytum borivilianum polysaccharide (CBP), as a dietary supplement administered at varying concentrations with feed (basal diet), on various cytokine-related responses in Labeo rohita fingerlings. Immune parameters and immune-related gene expressions were measured at 3rd, 4th, and 5th week after feeding. The results revealed that dietary administration of CBP at 0.2% and 0.4% for 4 weeks significantly upregulated serum lysozyme and phagocytic activity. Complement C3 and respiratory burst activity (RBA) were significantly higher after 4 weeks of CBP feeding. The immune related genes IL-8, IL-1β, TNF-α, and iNOS were downregulated (P < 0.05) in groups with 0.2% and 0.4% CBP supplemented diets at week 4. Expression of anti-inflammatory cytokines (IL-10 and TGF-β) was also downregulated (P < 0.5) after 4 weeks of feeding with 0.2% to 0.8% CBP. However, five weeks of CBP administration had no significant effect on immune gene expression, except TNF-α and IL-8. Fish fed with 0.4% CBP for 4 weeks showed maximum resistance against Aeromonas hydrophila (73.3% survival) compared to control. From these results, we recommend that CBP administration at 0.4% for 4 weeks could effectively improve immune response and disease resistance in L. rohita. PMID:26649325

  15. Inhibitory activity of monoacylglycerols on biofilm formation in Aeromonas hydrophila, Streptococcus mutans, Xanthomonas oryzae, and Yersinia enterocolitica.

    PubMed

    Ham, Youngseok; Kim, Tae-Jong

    2016-01-01

    Biofilm provides a bacterial hiding place by forming a physical barrier and causing physiological changes in cells. The elimination of biofilm is the main goal of hygiene. Chemicals that are inhibitory to biofilm formation have been developed for use in food, personal hygiene products, and medical instruments. Monoacylglycerols are recognized as safe and are used in food as emulsifiers. In this study, the inhibitory activity of monoacylglycerols on bacterial biofilm formation was evaluated systematically with four bacterial strains, Aeromonas hydrophila, Streptococcus mutans, Xanthomonas oryzae, and Yersinia enterocolitica. Monoacylglycerols with two specific lengths of fatty acid moiety, monolaurin and monobehenin, were found to have strong inhibitory activity toward bacterial biofilm formation of S. mutans, X. oryzae, and Y. enterocolitica in a strain specific manner. First, this result suggested that biofilm formation was not inhibited by the detergent characteristics of monoacylglycerols. This suggestion was supported by the inhibitory action of monolaurin on biofilm development but not on the initial cell attachment of Y. enterocolitica in flow cytometric observation. Second, it was also suggested that two distinct response mechanisms to monoacylglycerols existed in bacteria. The existence of these two inhibitory response mechanisms was bacterial strain specific.

  16. Effect of Withania somnifera (L. Dunal) root as a feed additive on immunological parameters and disease resistance to Aeromonas hydrophila in Labeo rohita (Hamilton) fingerlings.

    PubMed

    Sharma, Arun; Deo, Ashutosh D; Riteshkumar, S Tandel; Chanu, Thongam Ibemcha; Das, Arabinda

    2010-09-01

    The present study evaluated the efficacy of dietary doses of Withania somnifera (Ashwagandha) root powder on immunological parameters and disease resistance against the bacterial pathogen Aeromonas hydrophila infections in Indian major carp, Labeo rohita fingerlings. Fishes were fed with dry diet containing 0 gkg(-1) (control), 1 gkg(-1) (T(1)), 2 gkg(-1) (T(2)) and 3 gkg(-1) (T(3)) W. somnifera root powder for 42 days. Immunological (NBT level, Phagocytic activity, total immunoglobulin and lysozyme activity) parameters of fishes were examined at 0 days, 14 days, 28 days and 42 days of feeding. Fishes were challenged with A. hydrophila 42 days post feeding and mortalities (%) were recorded over 14 days post-infection. The results demonstrate that fishes fed with W. somnifera root showed enhanced NBT level, Phagocytic activity, total Immunoglobulin level and lysozyme activity (p<0.05) compared with the control group. The survivability was higher in experimental diets than the control group. Dietary W. somnifera at the level of 2 gkg(-1) showed significantly (P<0.05) higher protection (RPS 42.85+/-0.65%) against A. hydrophila infection than control. The results suggest that the W. somnifera root powder have a stimulatory effect on immunological parameters and increases disease resistance in L. rohita fingerlings against A. hydrophila infection.

  17. Genomic Characterization of the Novel Aeromonas hydrophila Phage Ahp1 Suggests the Derivation of a New Subgroup from phiKMV-Like Family

    PubMed Central

    Wang, Jian-Bin; Lin, Nien-Tsung; Tseng, Yi-Hsiung; Weng, Shu-Fen

    2016-01-01

    Aeromonas hydrophila is an opportunistic pathogenic bacterium causing diseases in human and fish. The emergence of multidrug-resistant A. hydrophila isolates has been increasing in recent years. In this study, we have isolated a novel virulent podophage of A. hydrophila, designated as Ahp1, from waste water. Ahp1 has a rapid adsorption (96% adsorbed in 2 min), a latent period of 15 min, and a burst size of 112 PFU per infected cell. At least eighteen Ahp1 virion proteins were visualized in SDS-polyacrylamide gel electrophoresis, with a 36-kDa protein being the predicted major capsid protein. Genome analysis of Ahp1 revealed a linear doubled-stranded DNA genome of 42,167 bp with a G + C content of 58.8%. The genome encodes 46 putative open reading frames, 5 putative phage promoters, and 3 transcriptional terminators. Based on high degrees of similarity in overall genome organization and among most of the corresponding ORFs, as well as phylogenetic relatedness among their DNAP, RNAP and major capsid proteins, we propose a new subgroup, designated Ahp1-like subgroup. This subgroup contains Ahp1 and members previously belonging to phiKMV-like subgroup, phiAS7, phi80-18, GAP227, phiR8-01, and ISAO8. Since Ahp1 has a narrow host range, for effective phage therapy, different phages are needed for preparation of cocktails that are capable of killing the heterogeneous A. hydrophila strains. PMID:27603936

  18. Effect of effluent from a nitrogen fertilizer factory and a pulp mill on the distribution and abundance of Aeromonas hydrophila in Albemarle Sound, North Carolina

    SciTech Connect

    Hazen, T.C.; Esch, G.W.

    1983-01-01

    The density of Aeromonas hydrophila, standard count bacteria, fecal coliform bacteria, and 18 physical and chemical parameters were measured simultaneously at six sites for 12 months in Albemarle Sound, N.C. One site was above and two sites were below the discharge plume of a Kraft pulping process paper mill. The fourth site was above and the remaining two sites were below the discharge point of a nitrogen fertilizer factory. The impact of the pulp mill on water quality was acute, whereas that of the nitrogen fertilizer factory was chronic and much more subtle. Diffusion chamber studies indicated that A. hydrophila survival is increased by pulp mill effluent and decreased by nitrogen fertilizer factor effluent. From correlation and regression analysis, A. hydrophila was found to be directly affected by phytoplankton density and, thus, indirectly by concentrations of phosphate, nitrate, and total organic carbon. These two point sources are suspect as indirect causes of red-sore disease epizootics, a disease of fish caused by A. hydrophila.

  19. Catecholamine-Stimulated Growth of Aeromonas hydrophila Requires the TonB2 Energy Transduction System but Is Independent of the Amonabactin Siderophore

    PubMed Central

    Dong, Yuhao; Liu, Jin; Pang, Maoda; Du, Hechao; Wang, Nannan; Awan, Furqan; Lu, Chengping; Liu, Yongjie

    2016-01-01

    The growth-stimulating effects of catecholamine stress hormones have been demonstrated in many pathogens. However, catecholamine-induced growth and its underlying mechanisms remain poorly understood in Aeromonas hydrophila. The present study sought to demonstrate that norepinephrine (NE), epinephrine (Epi), dopamine (Dopa), and L-dopa stimulate the growth of A. hydrophila in iron-restricted media containing serum. NE exhibited the strongest growth stimulation, which could be blocked by adrenergic antagonists. Furthermore, it was demonstrated that NE could sequester iron from transferrin, thereby providing a more accessible iron source for utilization by A. hydrophila. The deletion of the amoA gene associated with amonabactin synthesis revealed that the amonabactin siderophore is not required for NE-stimulated growth. However, the deletion of the TonB2 energy transduction system resulted in the loss of growth promotion by NE, indicating that a specific TonB-dependent outer membrane receptor might be involved in the transport of iron from transferrin. Collectively, our data show that catecholamine sensing promotes the growth of A. hydrophila in a manner that is dependent on the TonB2 energy transduction system. PMID:28018865

  20. Effect of effluent from a nitrogen fertilizer factory and a pulp mill on the distribution and abundance of Aeromonas hydrophila in Albemarle Sound, North Carolina.

    PubMed Central

    Hazen, T C; Esch, G W

    1983-01-01

    The density of Aeromonas hydrophila, standard count bacteria, fecal coliform bacteria, and 18 physical and chemical parameters were measured simultaneously at six sites for 12 months in Albemarle Sound, N.C. One site was above and two sites were below the discharge plume of a Kraft pulping process paper mill. The fourth site was above and the remaining two sites were below the discharge point of a nitrogen fertilizer factory. The impact of the pulp mill on water quality was acute, whereas that of the nitrogen fertilizer factory was chronic and much more subtle. Diffusion chamber studies indicated that A. hydrophila survival is increased by pulp mill effluent and decreased by nitrogen fertilizer factory effluent. From correlation and regression analysis, A. hydrophila was found to be directly affected by phytoplankton density and, thus, indirectly by concentrations of phosphate, nitrate, and total organic carbon. These two point sources are suspect as indirect causes of red-sore disease epizootics, a disease of fish caused by A. hydrophila. PMID:6297393

  1. DNA adenine methyltransferase (Dam) controls the expression of the cytotoxic enterotoxin (act) gene of Aeromonas hydrophila via tRNA modifying enzyme-glucose-inhibited division protein (GidA).

    PubMed

    Erova, Tatiana E; Kosykh, Valeri G; Sha, Jian; Chopra, Ashok K

    2012-05-01

    Aeromonas hydrophila is both a human and animal pathogen, and the cytotoxic enterotoxin (Act) is a crucial virulence factor of this bacterium because of its associated hemolytic, cytotoxic, and enterotoxic activities. Previously, to define the role of some regulatory genes in modulating Act production, we showed that deletion of a glucose-inhibited division gene (gidA) encoding tRNA methylase reduced Act levels, while overproduction of DNA adenine methyltransferase (Dam) led to a concomitant increase in Act-associated biological activities of a diarrheal isolate SSU of A. hydrophila. Importantly, there are multiple GATC binding sites for Dam within an upstream sequence of the gidA gene and one such target site in the act gene upstream region. We showed the dam gene to be essential for the viability of A. hydrophila SSU, and, therefore, to better understand the interaction of the encoding genes, Dam and GidA, in act gene regulation, we constructed a gidA in-frame deletion mutant of Escherichia coli GM28 (dam(+)) and GM33 (∆dam) strains. We then tested the expressional activity of the act and gidA genes by using a promoterless pGlow-TOPO vector containing a reporter green fluorescent protein (GFP). Our data indicated that in GidA(+) strains of E. coli, constitutive methylation of the GATC site(s) by Dam negatively regulated act and gidA gene expression as measured by GFP production. However, in the ∆gidA strains, irrespective of the presence or absence of constitutively active Dam, we did not observe any alteration in the expression of the act gene signifying the role of GidA in positively regulating Act production. To determine the exact mechanism of how Dam and GidA influence Act, a real-time quantitative PCR (RT-qPCR) assay was performed. The analysis indicated an increase in gidA and act gene expression in the A. hydrophila Dam-overproducing strain, and these data matched with Act production in the E. coli GM28 strain. Thus, the extent of DNA methylation

  2. Use of lupin, Lupinus perennis, mango, Mangifera indica, and stinging nettle, Urtica dioica, as feed additives to prevent Aeromonas hydrophila infection in rainbow trout, Oncorhynchus mykiss (Walbaum).

    PubMed

    Awad, E; Austin, B

    2010-05-01

    Feeding rainbow trout, Oncorhynchus mykiss (Walbaum), with 1% lupin, Lupinus perennis, mango, Mangifera indica, or stinging nettle, Urtica dioica, for 14 days led to reductions in mortality after challenge with Aeromonas hydrophila. In addition, there was significant enhancement in serum bactericidal activity, respiratory burst and lysozyme activity in the treatment groups compared to the controls. Use of lupin and mango led to the highest number of red blood and white blood cells in recipient fish, with use of stinging nettle leading to the highest haematocrit and haemoglobin values; the highest value of mean corpuscular volume and haemoglobin was in the control groups and those fed with stinging nettle.

  3. Acute cold stress improved the transcription of pro-inflammatory cytokines of Chinese soft-shelled turtle against Aeromonas hydrophila.

    PubMed

    Zhang, Zuobing; Chen, Bojian; Yuan, Lin; Niu, Cuijuan

    2015-03-01

    Chinese soft-shelled turtle, Pelodiscus sinensis, is widely cultured in East and Southeast Asian countries. It frequently encounters the stress of abrupt temperature changes, which leads to mass death in most cases. However, the mechanism underlying the stress-elicited death remains unknown. We have suspected that the stress impaired the immune function of Chinese soft-shelled turtle, which could result in the mass death, as we noticed that there was a clinical syndrome of infection in dead turtles. To test our hypothesis, we first performed bioinformatic annotation of several pro-inflammatory molecules (IL-1β, TNFα, IL-6, IL-12β) of Chinese soft-shelled turtle. Then, we treated the turtles in six groups, injected with Aeromonas hydrophila before acute cold stress (25 °C) and controls, after acute cold stress (15 °C) and controls as well as after the temperature was restored to 25 °C and controls, respectively. Subsequently, real-time PCR for several pro-inflammatory cytokines (IL-1β, TNFα, IL-6, IL-12β, IL-8 and IFNγ) was performed to assess the turtle immune function in spleen and intestine, 24 hours after the injection. We found that the mRNA expression levels of the immune molecules were all enhanced after acute cold stress. This change disappeared when the temperature was restored back to 25 °C. Our results suggest that abrupt temperature drop did not suppress the immune function of Chinese soft-shelled turtle in response to germ challenge after abrupt temperature drop. In contrast, it may even increase the expression of various cytokines at least, within a short time after acute cold stress.

  4. Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity.

    PubMed

    Sha, Jian; Kozlova, E V; Chopra, A K

    2002-04-01

    Three enterotoxins from the Aeromonas hydrophila diarrheal isolate SSU have been molecularly characterized in our laboratory. One of these enterotoxins is cytotoxic in nature, whereas the other two are cytotonic enterotoxins, one of them heat labile and the other heat stable. Earlier, by developing an isogenic mutant, we demonstrated the role of a cytotoxic enterotoxin in causing systemic infection in mice. In the present study, we evaluated the role of these three enterotoxins in evoking diarrhea in a murine model by developing various combinations of enterotoxin gene-deficient mutants by marker-exchange mutagenesis. A total of six isogenic mutants were prepared in a cytotoxic enterotoxin gene (act)-positive or -negative background strain of A. hydrophila. We developed two single knockouts with truncation in either the heat-labile (alt) or the heat-stable (ast) cytotonic enterotoxin gene; three double knockouts with truncations of genes encoding (i) alt and ast, (ii) act and alt, and (iii) act and ast genes; and a triple-knockout mutant with truncation in all three genes, act, alt, and ast. The identity of these isogenic mutants developed by double-crossover homologous recombination was confirmed by Southern blot analysis. Northern and Western blot analyses revealed that the expression of different enterotoxin genes in the mutants was correspondingly abrogated. We tested the biological activity of these mutants in a diet-restricted and antibiotic-treated mouse model with a ligated ileal loop assay. Our data indicated that all of these mutants had significantly reduced capacity to evoke fluid secretion compared to that of wild-type A. hydrophila; the triple-knockout mutant failed to induce any detectable level of fluid secretion. The biological activity of selected A. hydrophila mutants was restored after complementation. Taken together, we have established a role for three enterotoxins in A. hydrophila-induced gastroenteritis in a mouse model with the greatest

  5. Identification of gyrB and rpoB gene mutations and differentially expressed proteins between a novobiocin-resistant Aeromonas hydrophila catfish vaccine strain and its virulent parent strain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sequence comparison between the full-length 2412 bp DNA gyrase subunit B (gyrB) gene of a novobiocin resistant Aeromonas hydrophila AH11NOVO vaccine strain and that of its virulent parent strain AH11P revealed 10 missense mutations. Similarly, sequence comparison between the full-length 4092 bp RNA ...

  6. Methanol extract of Nyctanthes arbortristis seeds enhances non-specific immune responses and protects Oreochromis mossambicus (Peters) against Aeromonas hydrophila infection.

    PubMed

    Kirubakaran, C John Wesly; Subramani, Parasuraman Aiya; Michael, R Dinakaran

    2016-04-01

    Immunostimulation using medicinal plant extracts is a promising approach for prevention and control of diseases with reference to sustainable fish farming. Oreochromis mossambicus, dubbed as aquatic chicken is a cultured fish worldwide and a laboratory model organism. Aeromonas hydrophila is one of the major bacterial pathogens in fish farming that causes huge loss to aquaculture industries. In this study, we investigated the efficacy of methanol extract of Nyctanthes arbortristis seeds on disease resistance of O. mossambicus against live virulent A. hydrophila. We also investigated its effect on the non-specific immune parameters such as serum lysozyme, myeloperoxidase, antiprotease and specific immune parameters in terms of specific serum antibody titres assayed by bacterial agglutination test. Our studies indicate that intra-peritoneal administration of 20mg/kg methanol extract increases the Relative Percent Survival (RPS) of O. mossambicus challenged with LD80 of A. hydrophila. Further, both non-specific and specific immune parameters were enhanced by the methanol extract. Further experiments at molecular levels in the laboratory and also efficacy testing at field level are essential before applying this plant product in aquaculture industry.

  7. A cytotoxicity and comparative antibacterial study on the effect of Zataria multiflora Boiss, Trachyspermum copticum essential oils, and Enrofloxacin on Aeromonas hydrophila

    PubMed Central

    Malekinejad, Hassan; Bazargani-Gilani, Behnaz; Tukmechi, Amir; Ebrahimi, Hadi

    2012-01-01

    Objective: this study designed to test the antibacterial potency of enrofloxacin (ENR) and essential oils from Zataria multiflora Boiss (ZEO) and Trachyspermum copticum (TEO) on Aeromonas hydrophila. Material and Methods: The antibacterial potency of test compounds was determined by several methods including the inhibition zone diameter determination, microbroth dilution method and colorimetric method of MTT. The cytotoxicity of test substances was assessed on Chinook salmon (Oncorhynchus tshawytscha) embryo (CHSE-214) cells. Results: Results showed that ENR and tested essential oils exert antibacterial effect against A. hydrophila. Moreover, ENR exerted the most potent antibacterial effect with MIC values of 62.5 ng/ml. The natural compounds of ZEO and TEO also showed antibacterial effects with rather high MIC values of 0.315 mg/ml, and 1.25 mg/ml, respectively. None of the tested substances showed toxicity on CHSE-24 cells. Conclusion: It is concluded that ZEO and TEO could be applied to prevent from A. hydrophila infection. Moreover, data also suggest that MTT method could be both cost- and time-effective and accurate method of MIC determination. PMID:25050249

  8. Detection of Aeromonas hydrophila DNA oligonucleotide sequence using a biosensor design based on Ceria nanoparticles decorated reduced graphene oxide and Fast Fourier transform square wave voltammetry.

    PubMed

    Jafari, Safiye; Faridbod, Farnoush; Norouzi, Parviz; Dezfuli, Amin Shiralizadeh; Ajloo, Davood; Mohammadipanah, Fatemeh; Ganjali, Mohammad Reza

    2015-10-01

    A new strategy was introduced for ssDNA immobilization on a modified glassy carbon electrode. The electrode surface was modified using polyaniline and chemically reduced graphene oxide decorated cerium oxide nanoparticles (CeO2NPs-RGO). A single-stranded DNA (ssDNA) probe was immobilized on the modified electrode surface. Fast Fourier transform square wave voltammetry (FFT-SWV) was applied as detection technique and [Ru(bpy)3](2+/3+) redox signal was used as electrochemical marker. The hybridization of ssDNA with its complementary target caused a dramatic decrease in [Ru(bpy)3](2+/3+) FFT-SW signal. The proposed electrochemical biosensor was able to detect Aeromonas hydrophila DNA oligonucleotide sequence encoding aerolysin protein. Under optimal conditions, the biosensor showed excellent selectivity toward complementary sequence in comparison with noncomplementary and two-base mismatch sequences. The dynamic linear range of this electrochemical DNA biosensor for detecting 20-mer oligonucleotide sequence of A. hydrophila was from 1 × 10(-15) to 1 × 10(-8) mol L(-1). The proposed biosensor was successfully applied for the detection of DNA extracted from A. hydrophila in fish pond water up to 0.01 μg mL(-1) with RSD of 5%. Besides, molecular docking was applied to consider the [Ru(bpy)3](2+/3+) interaction with ssDNA before and after hybridization.

  9. Identification and characterization of a novel conserved 46 kD maltoporin of Aeromonas hydrophila as a versatile vaccine candidate in European eel (Anguilla anguilla).

    PubMed

    Feng, Jianjun; Lin, Peng; Guo, Songlin; Jia, Yuanyuan; Wang, Yilei; Zadlock, Frank; Zhang, Ziping

    2017-03-06

    European eel (Anguilla anguilla) is a crucial economic fish that has been plagued by Aeromonas hydrophila infections for many years. Vaccines that are cross-protective against multiple serotypes could provide an effective control against A. hydrophila-mediated diseases. The outer membrane proteins (OMPs) are highly immunogenic and capable of eliciting protective immune responses. This study reports the identification of a novel 46 kD maltoporin that is a conserved protective antigen for different serotypes of A. hydrophila. First, this study purified OMPs from the strains of A. hydrophila B10, B11, B12, B15, B19, and B20. Western blot analysis revealed that the 46 kD maltoporin of B11 could be strongly reacted with all the specific European eel antisera against the above OMPs from different serotypes A. hydrophila. Cloning and sequencing of the maltoporin revealed that it contains an open reading frame (ORF) of 1281 nucleotides encoding 426 amino acids. Further sequence alignment analysis using the NCBI Conserved Domain Database (CDD) along with performing three-dimensional structure analysis showed that this protein belongs to maltoporin family. Three different study groups of European eels were intraperitoneal injected with one of the following conditions: phosphate-buffered saline (PBS group), formaline-killed-whole-cell (FKC) of A. hydrophila (FKC group) or with the recombinant maltoporin (OMP group) to analyze the immunogenicity of the recombinant maltoporin purified by nickel chelate affinity chromatography. On 14, 21, 28 and 42 days post-vaccination respectively, proliferation of the whole blood cells, titers of specific antibody, and lysozyme activities of experimental eels were detected. On 28d post-vaccination, eels from the three groups were challenged by intraperitoneal injection with five different live strains of A. hydrophila (B10, B11, B15, B19, and B20). The results showed that the proliferation of whole blood cells in the OMP group was

  10. Bio-reduction of Fe(III) ores using three pure strains of Aeromonas hydrophila, Serratia fonticola and Clostridium celerecrescens and a natural consortium.

    PubMed

    García-Balboa, C; Bedoya, I Chion; González, F; Blázquez, M L; Muñoz, J A; Ballester, A

    2010-10-01

    The present work describes a research approach to the anaerobic bioleaching of Fe(III) ores. Three strains (Serratia fonticola, Aeromonas hydrophila and Clostridium celerecrescens) isolated from an acidic abandoned mine were selected to test their ability to reduce dissimilatory Fe(III). Total iron bio-reduction was achieved after 48 h using either the consortium or the Aeromonas cultures. In the latter case, there was no evidence of precipitates and Fe(II) remained in solution at neutral pH through complex formation with citrate. None of the other cultures tested (mixed culture and the two isolates) exhibited this behaviour. Biotechnologically, this is a very promising result since it obviates the problem associated with undesirable precipitation of iron compounds in Fe(III)-reducing bacterial cultures. The performance of the Aeromonas culture was improved progressively by adaptation to moderately acidic pH values (up to 4.5) and to three different Fe(III)-oxyhydroxides as the sole source of iron: ferrihydrite, hematite and jarosite, commonly found as weathering compounds at mine sites. Dissimilatory Fe(III)-reducers for iron extraction from ores is therefore especially attractive in that acidification of the surrounding area can be minimized.

  11. Isolation of potential probiotic Bacillus spp. and assessment of their subcellular components to induce immune responses in Labeo rohita against Aeromonas hydrophila.

    PubMed

    Ramesh, Dharmaraj; Vinothkanna, Annadurai; Rai, Amit Kumar; Vignesh, Venkada Subramanian

    2015-08-01

    Bacillus species isolated from the gut of healthy Labeo rohita (Hamilton) were screened for antibacterial activity against selected fish pathogens. Among the isolates, KADR5 and KADR6 showed antibacterial activity, tolerated low pH and high bile concentrations and were susceptibility to various antibiotics. Based on morphological and biochemical tests and 16S rRNA gene analysis the probiotic strains KADR5 and KADR6 were identified as Bacillus licheniformis and Bacillus pumilus, respectively. The immune stimulatory effect of subcellular components of probiotic Bacillus licheniformis KADR5 and Bacillus pumilus KADR6 in L. rohita against Aeromonas hydrophila infection was studied. Fish were immunized intraperitoneally in case of subcellular components [cell wall proteins (CWPs), extracellular proteins (ECPs), whole cell proteins (WCPs)] and orally in case of live cells (10(8) CFU/g of feed). After 14th day of administration, fishes from each group were challenged intraperitoneally with 0.1 ml of A. hydrophila cell suspension in PBS (10(5) cells ml(-1)). Groups immunized with subcellular components and live cells had significantly lower mortalities of 20-40% and 23-33%, respectively in comparison to control (80% mortality). The non specific immune factors in the cellular components and viable cells of the probiotics increased the expression of lysozyme and respiratory burst. Use of WCPs and CWPs resulted in better protection against A. hydrophila in L. rohita. Our results clearly reflect the potential of cellular components of the probiotics Bacillus species for the protection of fish against A. hydrophila infection by enhancing the immune response.

  12. In vivo bactericidal effect of Melaleuca alternifolia essential oil against Aeromonas hydrophila: Silver catfish (Rhamdia quelen) as an experimental model.

    PubMed

    F Souza, Carine; Baldissera, Matheus D; A Vaucher, Rodrigo; Lopes, Leonardo Q S; Vizzotto, Bruno S; Raffin, Renata P; Santos, Roberto C V; L da Veiga, Marcelo; U M da Rocha, Maria Izabel; Stefani, Lenita M; Baldisserotto, Bernardo

    2016-09-01

    Aeromonas hydrophila is one of the main causative agent of high mortality and significative economic losses in aquaculture and has become increasingly resistant to conventional antibiotics. One feasible alternative to control and treat it is the use of essential oils. This study aimed to evaluate A. hydrophila susceptibility to tea tree oil (TTO-Melaleuca alternifolia) in vivo, and the effect of this treatment. In vivo tests were performed using silver catfish (Rhamdia quelen) as the experimental model. Silver catfish were treated with TTO at 25 and 50 μL/L for seven days before infection. After seven days, the fish were inoculated with A. hydrophila via intramuscularly. Treatment with TTO at 50 μL/L was able to extend longevity of infected fish, and showed 88% of therapeutic success, even though it did not show curative efficacy. TTO treatment was not toxic under these tested concentrations, since biomarkers of hepatic and renal functions were not affected, and the concentration of 50 μL/L was able to prevent increased levels of aspartate aminotransferase. There was no significative differences regarding hematological parameters (p < 0.05). Treatment with TTO 50 μL/L was able to reduce histopathological alterations usually caused by this type of bacteria in the gills, but it was unable to reduce hepatic histopathological alterations. Our results showed, for the first time, that TTO has high activity against A. hydrophila and proved to be a natural alternative to prevent and control this pathogen.

  13. Role of Calmodulin-Calmodulin Kinase II, cAMP/Protein Kinase A and ERK 1/2 on Aeromonas hydrophila-Induced Apoptosis of Head Kidney Macrophages

    PubMed Central

    Banerjee, Chaitali; Khatri, Preeti; Raman, Rajagopal; Bhatia, Himanshi; Datta, Malabika; Mazumder, Shibnath

    2014-01-01

    The role of calcium (Ca2+) and its dependent protease calpain in Aeromonas hydrophila-induced head kidney macrophage (HKM) apoptosis has been reported. Here, we report the pro-apoptotic involvement of calmodulin (CaM) and calmodulin kinase II gamma (CaMKIIg) in the process. We observed significant increase in CaM levels in A. hydrophila-infected HKM and the inhibitory role of BAPTA/AM, EGTA, nifedipine and verapamil suggested CaM elevation to be Ca2+-dependent. Our studies with CaM-specific siRNA and the CaM inhibitor calmidazolium chloride demonstrated CaM to be pro-apoptotic that initiated the downstream expression of CaMKIIg. Using the CaMKIIg-targeted siRNA, specific inhibitor KN-93 and its inactive structural analogue KN-92 we report CaM-CaMKIIg signalling to be critical for apoptosis of A. hydrophila-infected HKM. Inhibitor studies further suggested the role of calpain-2 in CaMKIIg expression. CaMK Kinase (CaMKK), the other CaM dependent kinase exhibited no role in A. hydrophila-induced HKM apoptosis. We report increased production of intracellular cAMP in infected HKM and our results with KN-93 or KN-92 implicate the role of CaMKIIg in cAMP production. Using siRNA to PKACA, the catalytic subunit of PKA, anti-PKACA antibody and H-89, the specific inhibitor for PKA we prove the pro-apoptotic involvement of cAMP/PKA pathway in the pathogenicity of A. hydrophila. Our inhibitor studies coupled with siRNA approach further implicated the role of cAMP/PKA in activation of extracellular signal-regulated kinase 1 and 2 (ERK 1/2). We conclude that the alteration in intracellular Ca2+ levels initiated by A. hydrophila activates CaM and calpain-2; both pathways converge on CaMKIIg which in turn induces cAMP/PKA mediated ERK 1/2 phosphorylation leading to caspase-3 mediated apoptosis of infected HKM. PMID:24763432

  14. High yield expression of an AHL-lactonase from Bacillus sp. B546 in Pichia pastoris and its application to reduce Aeromonas hydrophila mortality in aquaculture

    PubMed Central

    2010-01-01

    Background Aeromonas hydrophila is a serious pathogen and can cause hemorrhagic septicemia in fish. To control this disease, antibiotics and chemicals are widely used which can consequently result in "superbugs" and chemical accumulation in the food chain. Though vaccine against A. hydrophila is available, its use is limited due to multiple serotypes of this pathogen and problems of safety and efficacy. Another problem with vaccination is the ability to apply it to small fish especially in high numbers. In this study, we tried a new way to attenuate the A. hydrophila infection by using a quorum quenching strategy with a recombinant AHL-lactonase expressed in Pichia pastoris. Results The AHL-lactonase (AiiAB546) from Bacillus sp. B546 was produced extracellularly in P. pastoris with a yield of 3,558.4 ± 81.3 U/mL in a 3.7-L fermenter when using 3-oxo-C8-HSL as the substrate. After purification with a HiTrap Q Sepharose column, the recombinant homogenous protein showed a band of 33.6 kDa on SDS-PAGE, higher than the calculated molecular mass (28.14 kDa). Deglycosylation of AiiAB546 with Endo H confirmed the occurrence of N-glycosylation. The purified recombinant AiiAB546 showed optimal activity at pH 8.0 and 20°C, exhibited excellent stability at pH 8.0-12.0 and thermal stability at 70°C, was firstly confirmed to be significantly protease-resistant, and had wide substrate specificity. In application test, when co-injected with A. hydrophila in common carp, recombinant AiiAB546 decreased the mortality rate and delayed the mortality time of fish. Conclusions Our results not only indicate the possibility of mass-production of AHL-lactonase at low cost, but also open up a promising foreground of application of AHL-lactonase in fish to control A. hydrophila disease by regulating its virulence. To our knowledge, this is the first report on heterologous expression of AHL-lactonase in P. pastoris and attenuating A. hydrophila virulence by co-injection with AHL

  15. Effect of salinity and incubation time of planktonic cells on biofilm formation, motility, exoprotease production, and quorum sensing of Aeromonas hydrophila.

    PubMed

    Jahid, Iqbal Kabir; Mizan, Md Furkanur Rahaman; Ha, Angela J; Ha, Sang-Do

    2015-08-01

    The aim of this study was to determine the effect of salinity and age of cultures on quorum sensing, exoprotease production, and biofilm formation by Aeromonas hydrophila on stainless steel (SS) and crab shell as substrates. Biofilm formation was assessed at various salinities, from fresh (0%) to saline water (3.0%). For young and old cultures, planktonic cells were grown at 30 °C for 24 h and 96 h, respectively. Biofilm formation was assessed on SS, glass, and crab shell; viable counts were determined in R2A agar for SS and glass, but Aeromonas-selective media was used for crab shell samples to eliminate bacterial contamination. Exoprotease activity was assessed using a Fluoro™ protease assay kit. Quantification of acyl-homoserine lactone (AHL) was performed using the bioreporter strain Chromobacterium violaceum CV026 and the concentration was confirmed using high-performance liquid chromatography (HPLC). The concentration of autoinducer-2 (AI-2) was determined with Vibrio harveyi BB170. The biofilm structure at various salinities (0-3 %) was assessed using field emission electron microscopy (FESEM). Young cultures of A. hydrophila grown at 0-0.25% salinity showed gradual increasing of biofilm formation on SS, glass and crab shell; swarming and swimming motility; exoproteases production, AHL and AI-2 quorum sensing; while all these phenotypic characters reduced from 0.5 to 3.0% salinity. The FESEM images also showed that from 0 to 0.25% salinity stimulated formation of three-dimensional biofilm structures that also broke through the surface by utilizing the chitin surfaces of crab, while 3% salinity stimulated attachment only for young cultures. However, in marked contrast, salinity (0.1-3%) had no effect on the stimulation of biofilm formation or on phenotypic characters for old cultures. However, all concentrations reduced biofilm formation, motility, protease production and quorum sensing for old culture. Overall, 0-0.25% salinity enhanced biofilm formation

  16. Molecular cloning of Japanese eel Anguilla japonica TNF-α and characterization of its expression in response to LPS, poly I:C and Aeromonas hydrophila infection

    NASA Astrophysics Data System (ADS)

    Feng, Jianjun; Guan, Ruizhang; Guo, Songlin; Lin, Peng; Zadlock, Frank

    2014-09-01

    As a potent pleiotropic cytokine, tumor necrosis factor-alpha (TNF-α) plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene ( Aj TNF-α) in the Japanese eel ( Anguilla japonica) were identified and characterized. The full-length AjTNF-α cDNA was 1 546 bp, including a 5'-untranslated region (UTR) of 13 bp, a 3'-UTR of 879 bp and an open reading frame of 654 bp encoding a protein of 218 amino acids. The full-length genomic sequence of AjTNF-α was 2 392 bp and included four exons and three introns. The putative AjTNF-α protein contained TNF family signature motifs, including a protease cleavage site, a transmembrane domain and two conserved cysteine residues. Quantitative real-time reverse transcription PCR analysis revealed AjTNF-α expression in a wide range of tissues, with predominant expression in blood and liver. Lower levels of expression were seen in spleen, gills, kidney, intestine, heart, and skin, with very low levels in muscle. The modulation of AjTNF-α expression after injection of eels with lipopolysaccharide (LPS), the viral mimic, poly I:C, or Aeromonas hydrophila was assessed in blood, liver, and kidney. In blood, TNF-α mRNA levels increased rapidly and then rapidly decreased after stimulation with LPS, poly I:C or A. hydrophila. However, the response to LPS and A. hydrophila peaked at 6 h while for poly I:C the peak was at 12 h. In liver, after injection with A. hydrophila, an up- and down-regulation of AjTNF-α expression occurred twice, peaking at 6 h and 24 h, respectively. No remarkable increase of AjTNF-α expression appeared in liver until 72 h after LPS or poly I:C treatment. In kidney, AjTNF-α expression increased significantly only at 72 h post-stimulation with LPS or A. hydrophila. Our results suggest that AjTNF-α plays an important role in fish in the defense against viral and bacterial infection.

  17. Peperomia pellucida leaf extract as immunostimulator in controlling motile aeromonad septicemia due to Aeromonas hydrophila in red hybrid tilapia, Oreochromis spp. farming

    PubMed Central

    Lee, S. W.; Sim, K. Y.; Wendy, W.; Zulhisyam, A. K.

    2016-01-01

    Aim: This study was revealed the potential of Peperomia pellucida leaf extract as an immunostimulator agent in controlling motile aeromonad septicemia due to Aeromonas hydrophila in red hybrid tilapia, Oreochromis sp. Materials and Methods: In the present study, minimum inhibitory concentration (MIC) of P. pellucida leaf extract against A. hydrophila was determined through two-fold microbroth dilution method. The plant extract was screening for its active compound using a gas chromatograph mass spectrometer, and the effectiveness of P. pellucida leaf extract as an immunostimulator agent was evaluated. The experimental fish were fed with medicated feed at three different concentrations (25 mg/kg, PP-25; 50 mg/kg, PP-50; and 100 mg/kg, PP-100) of P. pellucida leaf extract for 1 week before they were intraperitoneally exposed to A. hydrophila. Enzyme-linked immunosorbent assay was carried out to determine the value of antibody response to A. hydrophila in fish from a group of fish that received medicated feed, and the percentage of total cumulative mortality of the experimental fish were observed at the end of the experiment. Results: The results showed that the major bioactive compound is phytol (40%), and the MIC value was 31.5 mg/L. The value of antibody response to A. hydrophila in fish from a group of fish which received medicated feed (PP-25, 0.128±0.014 optical density [OD]; PP-50, 0.132±0.003 OD; and PP-100, 0.171±0.02 OD) was found significantly higher (p<0.05) compared to fish did not receive medicated feed (0.00 OD). Whereas, percentage cumulative mortality of fish from all groups of fish received medicated feed (PP-25, 18.0±3.2%; PP-50, 18.2±2.8%; and PP-100, 17.7±1.8%) were found significantly lower (p<0.05) compared to a group of fish did not receive medicated feed (83.2±1.4%). Conclusion: The findings of the present study indicated the huge potential of P. pellucida leaf extract as natural immunostimulator agent for aquaculture uses. PMID

  18. A Rare Case of Aeromonas Hydrophila Catheter Related Sepsis in a Patient with Chronic Kidney Disease Receiving Steroids and Dialysis: A Case Report and Review of Aeromonas Infections in Chronic Kidney Disease Patients

    PubMed Central

    Khalil, Muhammad Abdul Mabood; Rehman, Abdur; Kashif, Waqar Uddin; Rangasami, Manickam; Tan, Jackson

    2013-01-01

    Aeromonas hydrophila (AH) is an aquatic bacterium. We present a case of fifty-five-year-old gentleman with chronic kidney disease (CKD) due to crescentic IgA nephropathy who presented to us with fever. He was recently pulsed with methyl prednisolone followed by oral prednisolone and discharged on maintenance dialysis through a double lumen dialysis catheter. Blood culture from peripheral vein and double lumen dialysis catheter grew AH. We speculate low immunity due to steroids and uremia along with touch contamination of dialysis catheter by the patient or dialysis nurse could have led to this rare infection. Dialysis catheter related infection by AH is rare. We present our case here and take the opportunity to give a brief review of AH infections in CKD patients. PMID:24558624

  19. Dietary fermentable fiber upregulated immune related genes expression, increased innate immune response and resistance of rainbow trout (Oncorhynchus mykiss) against Aeromonas hydrophila.

    PubMed

    Yarahmadi, Peyman; Kolangi Miandare, Hamed; Farahmand, Hamid; Mirvaghefi, Alireza; Hoseinifar, Seyed Hossein

    2014-12-01

    This trial was carried out to investigate the effects of dietary administration of Vitacel(®), a commercial fermentable fiber, on immune related genes (Lysozyme, TNFα and HSP70) expression, innate immune response and resistance of rainbow trout against Aeromonas hydrophila. 120 healthy rainbow trout (81.65 ± 1.49 g) were distributed in six fiberglass tanks assigned to two treatments. The treatments were feeding rainbow trout with diets supplemented with 0 (control) or 10 g kg(-1) Vitacel(®) for 45 days. The results revealed that administration of fermentable fiber significantly (P < 0.05) upregulated lysozyme and TNFα gene expression. HSP70 gene expression was significantly lower in Vitacel(®) fed fish at the end of trial (P < 0.05). Furthermore dietary administrations of Vitacel(®) remarkably elevated rainbow trout innate immune parameters include serum lysozyme, ACH50, bactericidal activity and agglutination antibody titer (P < 0.05). Administration of 10 g kg(-1) Vitacel(®) significantly increased rainbow trout resistance against A. hydrophila (P < 0.05). The results of present study revealed that dietary Vitacel(®) can upregulates immune related genes expression and elevates innate immune response and disease resistance of rainbow trout.

  20. Effects of hirami lemon, Citrus depressa Hayata, leaf meal in diets on the immune response and disease resistance of juvenile barramundi, Lates calcarifer (bloch), against Aeromonas hydrophila.

    PubMed

    Shiu, Ya-Li; Lin, Hsueh-Li; Chi, Chia-Chun; Yeh, Shinn-Pyng; Liu, Chun-Hung

    2016-08-01

    The present study was conducted to evaluate the dietary supplementation of leaf meal from Citrus depressa Hayata on the growth, innate immune response, and disease resistance of juvenile barramundi, Lates calcarifer. Four diets were formulated to contain 0% (control), 1% (C1), 3% (C3), and 5% (C5) leaf meal, respectively. During a 56 d feeding trial, fish survival, growth performance, and feed efficiency were not significantly different among all groups. For immune response, respiratory burst, superoxide dismutase and lysozyme activities were not significantly different among all groups. However, fish fed the C5 diet for 56 d had significantly higher phagocytic activity. Also, fish fed C3 and C5 diets had significantly higher Mx gene expressions in spleens and head kidneys with nerve necrosis virus injections after 24 h. Disease resistance against Aeromonas hydrophila was increased by the C5 diet. In this study, barramundi fed on a diet containing 5% C. depressa Hayata leaf meal had significantly better innate immune response and disease resistance against A. hydrophila.

  1. Dietary supplementation with xylanase-expressing B. amyloliquefaciens R8 improves growth performance and enhances immunity against Aeromonas hydrophila in Nile tilapia (Oreochromis niloticus).

    PubMed

    Saputra, Febriyansyah; Shiu, Ya-Li; Chen, Yo-Chia; Puspitasari, Asthervina Widyastami; Danata, Ridha Handriany; Liu, Chun-Hung; Hu, Shao-Yang

    2016-11-01

    Bacillus amyloliquefaciens has attracted attention as a probiotic in aquaculture due to its immunostimulatory activity against pathogenic infection. Xylanases are extensively used in animal feed to degrade plant ingredients, enhancing nutrient utilization and increasing the growth rate of various animals. In the present study, the effects of dietary supplementation with B. amyloliquefaciens and xylanase-expressing B. amyloliquefaciens R8 on the growth of Nile tilapia (Oreochromis niloticus) and immunity against Aeromonas hydrophila were evaluated. The results showed that the xylanase activity in the intestine, weight gain (WG), feed efficiency (FE) and condition factor (CF) of Nile tilapia fed B. amyloliquefaciens R8 for 2 months were significantly increased compared with those of the fish fed the control diet and B. amyloliquefaciens. Moreover, the mRNA expression of growth- and metabolism-related genes, such as insulin-like growth factor-1 (igf-1), glucokinase (GK), glucose-6-phosphate 1-dehydrogenase (G6PD), and glucose-6-phosphatase (G6Pase), was significantly induced in Nile tilapia fed administered B. amyloliquefaciens R8, and this group also exhibited a higher survival rate than the control fish following a challenge with A. hydrophila. The phagocytic activity and respiratory burst activity of head kidney leukocytes as well as the serum lysozyme activity of B. amyloliquefaciens R8-fed Nile tilapia were significantly higher than those of fish fed the control diet for 2 months. Superoxide dismutase (SOD) levels in the head kidney leukocytes of Nile tilapia fed B. amyloliquefaciens R8 differed from those of fish fed the control diet, but this was not significant. These results indicate that dietary supplementation with xylanase-expressing B. amyloliquefaciens R8 improves growth performance and enhances immunity and disease resistance against A. hydrophila in Nile tilapia.

  2. Apolipoprotein A1 in channel catfish: Transcriptional analysis, antimicrobial activity, and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were to: 1) determine transcriptional profiles of apolipoprotein A1 (ApoA1) in collected channel catfish tissues after infection with A. hydrophila by bath immersion; 2) investigate whether recombinant channel catfish apolipoprotein A1 produced in E. coli expression syst...

  3. Significant association between SNPs in the superoxide dismutase 3, extracellular (SOD3) gene and resistance to Aeromonas hydrophila in the freshwater mussel Hyriopsis cumingii.

    PubMed

    Wang, G L; Li, X L; Li, J L

    2013-12-01

    Extracellular superoxide dismutase (SOD3) is a major antioxidant enzyme that protects organs from damage by reactive oxygen species (ROS). In this study, the SOD3 gene was identified and characterized from the freshwater mussel Hyriopsis cumingii (Hc-SOD3). The cDNA sequence consists of 763 bp, encoding a protein of 208 amino acids. The amino acid sequence possesses two CuZnSOD signature sequences, and amino acids required for binding of Cu (His-93, -95, -110 and -169) and Zn (His-110, -118, -129 and Asp-132) were conserved in Hc-SOD3. The Hc-SOD3 genomic sequence was 9165 bp in length, containing four exons and three introns. Eighteen single nucleotide polymorphisms were detected in the Hc-SOD3 gene from resistant stock (RS) and susceptible stock (SS) of H. cumingii to Aeromonas hydrophila. The genotype and allele distribution were examined in resistant and susceptible stocks. Among them, a C/G substitution at the g.7994C>G locus and G/C substitution at the g.8087G>C locus were significantly associated with resistance/susceptibility of H. cumingii to A. hydrophila, both in genotype (P = 0.017, P = 0.004 respectively) and allele frequency (P = 0.021, P = 0.006 respectively). Linkage disequilibrium analysis revealed that g.7994C>G, g.8001A>G, g.8035G>A, g.8087G>C and g.8191T>A were in linkage disequilibrium. The results suggest that the two polymorphic loci, g.7994C>G and g.8087G>C, could be potential genetic markers for future molecular selection of strains that are resistant to diseases.

  4. Dietary supplementation with Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus oryzae enhance immunity and disease resistance against Aeromonas hydrophila and Streptococcus iniae infection in juvenile tilapia Oreochromis niloticus.

    PubMed

    Iwashita, Marina Keiko P; Nakandakare, Ivan B; Terhune, Jeffery S; Wood, Theresa; Ranzani-Paiva, Maria José T

    2015-03-01

    A feeding trial was conducted to investigate the effects of dietary administration of probiotic with Bacillus subtilis, Aspergillus oryzae and Saccharomyces cerevisiae on growth, innate immune response, Hemato-immunological parameters and disease resistance of Nile tilapia, Oreochromis niloticus. Animals were distributed in three equal groups, each of five replicates and received one of the following experimental diets for four weeks: Control, non-supplemented diet; 5 g kg(-1) probiotic mixture (B. subtilis 1.5 × 10(9) CFU g(-1), S. cerevisiae 10(9) CFU g(-1) and A. oryzae 2 × 10(9) CFU g(-1)); and 10 g kg(-1) probiotic mixture (B. subtilis 3.0 × 10(9) CFU g(-1), S. cerevisiae 2.0 × 10(9) CFU g(-1) and A. oryzae 4.0 × 10(9) CFU g(-1)). The respiratory burst activity, white blood cells and hematological parameters were evaluated after four, five and six weeks of feeding. At the end of the growth trial, fish were sampled for intestinal microbiology and challenged by intraperitoneal injection of LD50 concentration of Aeromonas hydrophila and Streptococcus iniae. Mortality was recorded for the following 3 weeks. Results showed that administration of the probiotic had no significant effect on the growth rates of Nile tilapias, although the fish fed probiotics had better feed conversion. Respiratory burst activity, erythrocyte fragility and levels of white blood cells were significantly improved in tilapias fed diet supplemented with probiotic levels (P < 0.05), which may exhibit up-regulating effects on tilapia immune parameters. The cumulative mortality after A. hydrophila and S. iniae challenge decreased in tilapias fed with probiotic (P < 0.05). The present study demonstrated the potential of B. subtilis, S. cerevisiae and A. oryzae combined as beneficial dietary probiotic in juvenile O. niloticus.

  5. Dietary protein enhances non-specific immunity, anti-oxidative capability and resistance to Aeromonas hydrophila in Labeo rohita fingerlings pre-exposed to short feed deprivation stress.

    PubMed

    Yengkokpam, Sona; Debnath, Dipesh; Sahu, N P; Pal, A K; Jain, K K; Baruah, Kartik

    2016-12-01

    Present experiment was conducted to study the effect of dietary protein levels on growth, immunity and anti-oxidative status of Labeo rohita fingerlings during feed deprivation followed by refeeding. Fish (5.44 ± 0.10 g) were deprived of feed for 3 weeks and then re-fed to satiation for 5 weeks with one of the diets containing 25 (25P), 30 (30P), 35 (35P) or 40 (40P) percent crude protein (CP) level. In addition to these groups, a control group (C) was also maintained by feeding to satiation level twice daily with a diet containing 30% CP throughout the experimental period. At the end of 8-weeks' trial, fish were challenged with Aeromonas hydrophila and survival was recorded for the next 7 days. Complete recovery of growth in terms of weight gain percentage was achieved in the fish fed 35 and 40% protein during refeeding. The body indices (condition factor and hepatosomatic index), haematological parameters and serum protein contents at the end of the experimental trial were not significantly different (P > 0.05) among different groups suggesting that the overall health of the fish was not compromised. However, respiratory burst activity and serum lysozyme activity were indicative of a better immune function in the higher protein fed groups (35P and 40P) than the lower protein groups (25P and 30P). Following challenge with Aeromonas hydrophila, survival rate, blood monocyte%, respiratory burst activity, serum lysozyme activity, serum protein and globulin were significantly higher (P < 0.05) in the 35P and 40P groups compared to the other groups. Further, fish fed lower dietary protein were not able to restore the activities of anti-oxidative enzymes (superoxide dismutase and catalase) in the liver. Conclusively, an improved disease resistance capability and immune status was observed in the fish fed a higher dietary protein (35-40%), even out-performing the daily-fed fish.

  6. Evaluation of the roles played by Hcp and VgrG type 6 secretion system effectors in Aeromonas hydrophila SSU pathogenesis.

    PubMed

    Sha, Jian; Rosenzweig, Jason A; Kozlova, Elena V; Wang, Shaofei; Erova, Tatiana E; Kirtley, Michelle L; van Lier, Christina J; Chopra, Ashok K

    2013-06-01

    Aeromonas hydrophila, a Gram-negative bacterium, is an emerging human pathogen equipped with both a type 3 and a type 6 secretion system (T6SS). In this study, we evaluated the roles played by paralogous T6SS effector proteins, hemolysin co-regulated proteins (Hcp-1 and -2) and valine glycine repeat G (VgrG-1, -2 and -3) protein family members in A. hydrophila SSU pathogenesis by generating various combinations of deletion mutants of the their genes. In addition to their predicted roles as structural components and effector proteins of the T6SS, our data clearly demonstrated that paralogues of Hcp and VgrG also influenced bacterial motility, protease production and biofilm formation. Surprisingly, there was limited to no observed functional redundancy among and/or between the aforementioned T6SS effector paralogues in multiple assays. Our data indicated that Hcp and VgrG paralogues located within the T6SS cluster were more involved in forming T6SS structures, while the primary roles of Hcp-1 and VgrG-1, located outside of the T6SS cluster, were as T6SS effectors. In terms of influence on bacterial physiology, Hcp-1, but not Hcp-2, influenced bacterial motility and protease production, and in its absence, increases in both of the aforementioned activities were observed. Likewise, VgrG-1 played a major role in regulating bacterial protease production, while VgrG-2 and VgrG-3 were critical in regulating bacterial motility and biofilm formation. In an intraperitoneal murine model of infection, all Hcp and VgrG paralogues were required for optimal bacterial virulence and dissemination to mouse peripheral organs. Importantly, the observed phenotypic alterations of the T6SS mutants could be fully complemented. Taking these results together, we have further established the roles played by the two known T6SS effectors of A. hydrophila by defining their contributions to T6SS function and virulence in both in vitro and in vivo models of infection.

  7. The role of three heat shock protein genes in the immune response to Aeromonas hydrophila challenge in marbled eel, Anguilla marmorata

    PubMed Central

    Liang, Fenfei; Zhang, Guosong; Wang, Li

    2016-01-01

    Heat shock proteins (HSPs) are highly conserved molecular chaperones that play critical roles in both innate and adaptive immunity. However, little information about HSPs from marbled eel Anguilla marmorata is known. In this study, the full-length Amhsp90 (2527 bp), Amhsp70 (2443 bp) and Amhsc70 (2247 bp) were first cloned from A. marmorata, using rapid amplification of cDNA ends, containing open reading frames of 2181, 1932 and 1950 bp in length, and encoding proteins with 726, 643 and 649 amino acids, respectively. The deduced amino acid sequences of three Amhsps shared a high homology similarity with other migratory fish. Real-time fluorescent quantitative polymerase chain reaction was used to evaluate tissue-specific distribution and mRNA expression levels of three Amhsps subjected to infection with Aeromonas hydrophila. The mRNA expression of three Amhsps in eight tested tissues, namely liver, heart, muscle, gill, spleen, kidney, brain and intestine, of juvenile A. marmorata was evaluated to reveal the major expression distribution in liver, intestine, muscle and heart. After pathogen challenge treatments, mRNA transcriptions of three Amhsps revealed a significant regulation at various time points in the same tissue. All these findings suggest that Amhsps may be involved in the immune response in A. marmorata. PMID:27853553

  8. Trigonella foenum-graceum (Seed) Extract Interferes with Quorum Sensing Regulated Traits and Biofilm Formation in the Strains of Pseudomonas aeruginosa and Aeromonas hydrophila

    PubMed Central

    Husain, Fohad Mabood; Ahmad, Iqbal; Khan, Mohd Shahnawaz; Al-Shabib, Nasser Abdulatif

    2015-01-01

    Trigonella foenum-graecum L. (Fenugreek) is an important plant of the Leguminosae family known to have medicinal properties. However, fraction based antiquorum sensing and antibiofilm activities have not been reported from this plant. In the present study T. foenum-graecum seed extract was sequentially fractionated and sub-MICs were tested for above activities. The methanol fraction of the extract demonstrated significant inhibition of AHL regulated virulence factors: protease, LasB elastase, pyocyanin production, chitinase, EPS, and swarming motility in Pseudomonas aeruginosa PAO1 and PAF79. Further, QS dependent virulence factor in the aquatic pathogen Aeromonas hydrophila WAF38 was also reduced. Application of T. foenum-graecum seed extract to PAO1, PAF79, and WAF38 decreased the biofilm forming abilities of the pathogens by significant levels. The extract also exhibited reduced AHL levels and subsequent downregulation of lasB gene. In vivo study showed an enhanced survival of PAO1-preinfected C. elegans after treatment with extract at 1 mg/mL. Further, the major compound detected by GC-MS, caffeine, reduced the production of QS regulated virulence factors and biofilm at 200 µg/mL concentration indicating its role in the activity of the methanol extract. The results of the present study reveal the potential anti-QS and antibiofilm property of T. foenum-graceum extract and caffeine. PMID:26000026

  9. In vitro antibacterial activities of ethanol extract of iranian propolis (EEIP) against fish pathogenic bacteria (Aeromonas hydrophila, Yersinia ruckeri & Streptococcus iniae).

    PubMed

    Tukmechi, Amir; Ownagh, Abdolghaffar; Mohebbat, Ali

    2010-10-01

    The "in vitro" antibacterial activity of ethanol extract of propolis (EEIP) from Urmia, Iran was investigated against three prevalent species of fish bacterial pathogens including: Aeromonas hydrophila LMG 3770, Yersinia ruckeri LMG 3279 and Streptococcus iniae LMG 14520. In this study two standard susceptibility testing techniques (Micro-broth dilution method and Agar-well diffusion method) were used to evaluation of the antibacterial activity of EEIP against the mentioned micro-organisms. Also the chemical composition of propolis was determined by the method of Gas chromatography-mass spectrometry (GC-MS). Twenty-six compounds were identified by gas chromatography-mass spectrometry analysis. Results showed Chemical composition of EEIP contained significant amounts of flavonoids, Sesquiterpenes - mainly Eudesmol and Caryophyllene oxide - aromatic acid, and low amounts of aldehydes and triterpens. Furthermore the ethanol extract of propolis inhibited the growth of all examined micro-organisms with the highest antimicrobial activity against Gram-positive bacteria Streptococcus iniae. Ethanol did not influence the antimicrobial effect of EEIP. These antibacterial properties would warrant further studies on the clinical applications of propolis in aquaculture field.

  10. Screening probiotic candidates for a mixture of probiotics to enhance the growth performance, immunity, and disease resistance of Asian seabass, Lates calcarifer (Bloch), against Aeromonas hydrophila.

    PubMed

    Lin, Hsueh-Li; Shiu, Ya-Li; Chiu, Chiu-Shia; Huang, Shih-Ling; Liu, Chun-Hung

    2017-01-01

    Six bacteria, including, Lactobacillus casei M15, Lac. plantarum D8, Lac. pentosus BD6, Lac. fermentum LW2, Enterococcus faecium 10-10, and Bacillus subtilis E20, and one yeast, Saccharomyces cerevisiae P13 were selected as probiotics for Asian seabass, Lates calcarifer, by tracking the growth performance and disease resistance of fish against Aeromonas hydrophila in the first trial. The probiotic efficiency screening results showed that B. subtilis E20 and Lac. pentosus BD6, and S. cerevisiae P13 and Lac. fermentum LW2 respectively improved either the growth performance or disease resistance. Therefore, these four probiotics were then selected to prepare a probiotics mixture, and this was incorporated in equal proportions into diets for Asian seabass at levels of 0 (control), and 10(6) (MD6), 10(7) (MD7), 10(8) (MD8), and 10(9) (MD9) colony-forming units (cfu) (kg diet)(-1). A synergistic effect of the combined probiotics was investigated in this study, and the probiotics mixture was able to improve both the growth performance and health status of fish. After 56 days of feeding, fish fed the MD9 diet had a higher final weight and percentage of weight gain. In addition, protein contents in the dorsal muscle of fish fed the MD8 and MD9 diets were significantly higher compared to the control. For the pathogen challenge test, fish fed the MD7, MD8, and MD9 diets had significantly lower cumulative mortalities after A. hydrophila infection compared to those of fish fed the control and MD6 diets, which might have been due to increased respiratory bursts, decreased superoxide dismutase activity in leucocytes, and increased phagocytic activity. Therefore, we considered that the probiotics mixture could adequately provide probiotic efficiency for Asian seabass, and the diet containing 10(9) cfu (kg diet)(-1) probiotic mixture is recommended to improve the growth and health status of Asian seabass.

  11. Parenteral immunization of PLA/PLGA nanoparticle encapsulating outer membrane protein (Omp) from Aeromonas hydrophila: Evaluation of immunostimulatory action in Labeo rohita (rohu).

    PubMed

    Rauta, Pradipta Ranjan; Nayak, Bismita

    2015-05-01

    Advanced vaccine research approaches needs to explore on biodegradable nanoparticles (NPs) based vaccine carrier that can serve as antigen delivery systems as well as immuno-stimulatory action to induce both innate and adaptive immune response in fish. Immunogenicity of PLA and PLGA NPs encapsulating outer membrane protein (Omp) antigen of Aeromonas hydrophila were evaluated through intra-peritoneal injection in fish, Labeo rohita. Antigen loaded PLA-Omp (223.5 ± 13.19 nm) and PLGA-Omp (166.4 ± 21.23 nm) NPs were prepared using double emulsion method by efficiently encapsulating the antigen reaching the encapsulation efficiency 44 ± 4.58% and 59.33 ± 5.13% respectively. Our formulated PLA Omp and PLGA-Omp NPs were in nanometer range (<500 nm) and could be successfully endocyted in the body. Despite low antigen loading in PLA-Omp, it showed considerably slower antigen release in vitro than PLGA-Omp NPs. Other physical properties like zetapotential values and poly dispersity index (PDI) confirmed the stability as well as monodisperse nature of the formulated nanoparticles. The spherical and isolated nature of PLA-Omp and PLGA-Omp NPs were revealed by SEM analysis. Upon immunization of all antigenic formulations (PLA-Omp NP, PLGA-Omp NP, FIA-Omp, PLA NP, PLGA NP, PBS as control), significant higher bacterial agglutination titre and haemolytic activity were observed in case of PLA-Omp and PLGA-Omp immunized groups than rest groups at both 21 days and 42 days. The specific antibody response was significantly increased and persisted up to 42 days of post immunization by PLA-Omp, PLGA-Omp, FIA-Omp. PLA-Omp NPs showed better immune response (higher bacterial agglutination titre, haemolytic activity, specific antibody titre, higher percent survival upon A. hydrophila challenge) than PLGA-Omp in L. rohita confirming its better efficacy. Comparable antibody response of PLA-Omp and PLGA-Omp with FIA-Omp treated groups suggested that PLA and PLGA could be replacement for

  12. Effect of guava leaves on the growth performance and cytokine gene expression of Labeo rohita and its susceptibility to Aeromonas hydrophila infection.

    PubMed

    Giri, Sib Sankar; Sen, Shib Sankar; Chi, Cheng; Kim, Hyoun Joong; Yun, Saekil; Park, Se Chang; Sukumaran, V

    2015-10-01

    The present study aimed to evaluate the effects of Psidium guajava L. (guava) leaves on the growth and immune response of the fish species Labeo rohita and its susceptibility to Aeromonas hydrophila infection. Diets containing five different concentrations of guava leaves (0% [basal diet], 0.1% [G1], 0.5% [G2], 1% [G3], and 1.5% [G4]) were fed to fish (average weight: 11.1 g) for 60 days. Various growth and immune parameters were examined 60 days post-feeding. Fish were challenged with A. hydrophila at the end of the trial, and mortalities were recorded over 15 days post-infection. We found that growth parameters such as percent weight gain (657.61 ± 9.74) and specific growth rate (3.37 ± 0.021) were significantly higher in G2 group than in the control (P < 0.05). Among the immune parameters examined, lysozyme levels (79.5 ± 5.1 U mL(-1)), leukocyte phagocytic activity (52 ± 4.3%), and alternative complement pathway activity (ACP) (186.1 ± 8.3 U mL(-1)) were significantly high (P < 0.05) in G2 fed group; there was, however, no significant effect of guava leaves at any concentration on plasma IgM level. Of the cytokine-related genes examined, interleukin-1beta (IL-1β) and tumour necrosis factor-alpha (TNF-α) were up-regulated in the head-kidney, intestine, and hepatopancreas of fish fed experimental diets, and expression was significantly higher in G2 and G3 than in the control group. In contrast, gene expression of IL-10, transforming growth factor-beta (TGF-β), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and transcription factor nuclear factor-κB (NF-κB) were down-regulated in the treatment groups. Moreover, fish fed the G2 diet exhibited a significantly higher post-challenge survival rate (66.66%). Collectively, these results suggest that dietary supplementation with guava leaves (at 0.5% concentration) could promote growth performance and strengthen immunity of L. rohita. Guava leaves therefore represent a promising feed additive

  13. MONITORING FOR AEROMONAS SPECIES AFTER TREATMENT WITH COMMON DRINKING WATER DISINFECTANTS

    EPA Science Inventory

    The sensitivity of Aeromonas spp. To free chlorine, chloramine and ultraviolet (UV) disinfection was determined. Aeromonas hydrophila is a contaminant listed on the USEPA's 1998 Contaminant Candidate List (CCL). Experiments using free chlorine indicated that the Aeromonas spp. ...

  14. Effect of dietary copper on the growth performance, non-specific immunity and resistance to Aeromonas hydrophila of juvenile Chinese mitten crab, Eriocheir sinensis.

    PubMed

    Sun, Shengming; Qin, Jianguang; Yu, Na; Ge, Xianping; Jiang, Haibo; Chen, Liqiao

    2013-05-01

    An 8-week feeding trial was conducted to determine the dietary copper (Cu) on growth performance and immune responses of juvenile Chinese mitten crab Eriocheir sinensis. Six semi-purified diets with six copper levels (1.88, 11.85, 20.78, 40.34, 79.56 and 381.2 mg kg(-1) diet) of CuSO4·5H2O were fed to E. sinensis (0.45 ± 0.01 g). Each diet was fed to the crab in five replicates. The crab fed diets with 20.78 and 40.34 mg Cu kg(-1) diet had significantly greater weight gain and hemolymph oxyhemocyanin content than those fed diets with 1.88 and 381.2 mg Cu kg(-1) diet. Survival rates of crab were not significantly different between all treatment groups. The activities of copper-zinc superoxide dismutase (Cu-Zn SOD), phenoloxidase (PO), and total hemocyte count (THC) significantly increased when the supplementation of dietary copper reached 20.78-40.34 mg Cu kg(-1) diets. In the bacteria challenge experiment with Aeromonas hydrophila, survival rates significantly increased and reached a plateau when the dietary copper increased from 1.88 to 40.34 mg kg(-1), whereas significantly decreased when the dietary copper increased from 40.34 to 381.2 mg kg(-1). This study indicates that the level of dietary copper is important in regulating growth and immune response in crab.

  15. Chitinases biosynthesis by immobilized Aeromonas hydrophila SBK1 by prawn shells valorization and application of enzyme cocktail for fungal protoplast preparation.

    PubMed

    Halder, Suman Kumar; Maity, Chiranjit; Jana, Arijit; Ghosh, Kuntal; Das, Arpan; Paul, Tanmay; Das Mohapatra, Pradeep Kumar; Pati, Bikas Ranjan; Mondal, Keshab Chandra

    2014-02-01

    Production and optimization of β-N-acetyl glucosaminidase and chitinase by Ca-alginate immobilized Aeromonas hydrophila SBK1 was carried out using prawn shell as cost-effective substrate. Beads prepared with 5.0% Na-alginate (containing 2.0% colloidal chitin) and 1.0 M CaCl2 showed considerable beads integrity and supported maximum production of chitinolytic enzymes. Bead diameter, 3 mm; temperature, 35°C; pH 7.0; agitation, 90 rpm were found ideal for the maximum production of the enzymes. The fermentation and thermodynamic indices revealed the feasibility of immobilized cells over free cells for enzymes production. Reasonable amount of chitosaccharides (degree of polymerization; 1-6) accumulated in the production media which have paramount antioxidant activity. Scale up experiment was successfully carried out in 5 L fermentor. In immobilized state, the chitosaccharides yield and antioxidant activity increased about 44.76% and 22.22%, whereas specific productivity of β-N-acetyl glucosaminidase and chitinase increased by 22.86% and 33.37% over free state. The cell entrapped beads can be reused upto ten cycles without marked loss of its biocatalytic efficiency. High level of protoplast of Aspergillus niger was generated by treating mycelia with 10 U/ml of crude chitinase after 4 h at pH 7.0 and in the temperature 35-40°C, and 67% of the protoplasts were found to be regenerated.

  16. Melaleuca alternifolia essential oil enhances the non-specific immune system and prevents oxidative damage in Rhamdia quelen experimentally infected by Aeromonas hydrophila: Effects on cholinergic and purinergic systems in liver tissue.

    PubMed

    Baldissera, Matheus D; Souza, Carine F; Júnior, Guerino B; de Vargas, Agueda C; Boligon, Aline A; de Campos, Marli M A; Stefani, Lenita M; Baldisserotto, Bernardo

    2017-02-01

    The aim of this study was to evaluate the effects of M. alternifolia essential oil used to treat silver catfish (Rhamdia quelen) experimentally infected by Aeromonas hydrophila on oxidative stress variables, and for the first time, on hepatic enzymes of the cholinergic and adenosinergic systems. For that, fish were divided into six groups (A-F), each containing seven animals. Groups A, B and C were composed of uninfected animals, while animals in groups D, E and F were intramuscularly inoculated with A. hydrophila. Groups B and E received a prophylactic bath with M. alternifolia essential oil (50 μL/L, diluted in ethanol) for seven days, while groups C and F were exposed to ethanol. After the prophylactic baths, groups D, E and F were inoculated with 100 μL of A. hydrophila solution (2.1 × 10(9) colony-forming unit). Two days after inoculation, the animals were euthanized and liver samples were collected. Infected animals (the group D) showed increased TBARS and protein carbonylation levels, while CAT, AChE and ADA activities decreased compared to uninfected animals (the group A). The prophylactic treatment with M. alternifolia essential oil (the group E) prevented the alterations caused by A. hydrophila, but it did not change AChE activity. Thus, the prophylactic treatment prevents damage caused by lipids and proteins, as well as alterations of the adenosinergic system, demonstrating that the anti-inflammatory effect of TTO is mediated by the adenosinergic pathway. In addition, TTO prophylactic treatment might be considered an important approach to prevent the hepatic damage caused by A. hydrophila.

  17. Effects of dietary administration of stinging nettle (Urtica dioica) on the growth performance, biochemical, hematological and immunological parameters in juvenile and adult Victoria Labeo (Labeo victorianus) challenged with Aeromonas hydrophila.

    PubMed

    Ngugi, Charles C; Oyoo-Okoth, Elijah; Mugo-Bundi, James; Orina, Paul Sagwe; Chemoiwa, Emily Jepyegon; Aloo, Peninah A

    2015-06-01

    We investigated effects of dietary administration of stinging nettle (Urtica dioica) on growth performance, biochemical, hematological and immunological parameters in juvenile and adult Victoria Labeo (Labeo victorianus) against Aeromonas hydrophila. Fish were divided into 4 groups and fed for 4 and 16 weeks with 0%, 1%, 2% and 5% of U. dioica incorporated into the diet. Use of U. dioica in the diet resulted in improved biochemical, hematological and immunological parameters. Among the biochemical parameters; plasma cortisol, glucose, triglyceride and cholesterol decreased while total protein and albumin in fish increased with increasing dietary inclusion of U. dioica. Among the haematology parameters: red blood cell (RBC), white blood cell (WBC) counts, haematocrit (Htc), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC) and netrophiles increased with increasing dietary inclusion levels of U. dioica, some depending on the fish age. Serum immunoglobulins, lysozyme activity and respiratory burst were the main immunological parameters in the adult and juvenile L. victorianus measured and they all increased with increasing herbal inclusion of U. dioica in the diet. Dietary incorporation of U. dioica at 5% showed significantly higher relative percentage survival (up to 95%) against A. hydrophila. The current results demonstrate that using U. dioica can stimulate fish immunity and make L. victorianus more resistant to bacterial infection (A. hydrophila).

  18. Effects of replacing soybean meal with rubber seed meal on growth, antioxidant capacity, non-specific immune response, and resistance to Aeromonas hydrophila in tilapia (Oreochromis niloticus × O. aureus).

    PubMed

    Deng, Junming; Mai, Kangsen; Chen, Liqiao; Mi, Haifeng; Zhang, Lu

    2015-06-01

    This study evaluated the effects of replacing soybean meal (SBM) with rubber seed meal (RSM) on growth, antioxidant capacity, non-specific immune response and resistance to Aeromonas hydrophila in tilapia (Oreochromis niloticus × Oreochromis aureus). Five experimental diets were formulated with 0 (control), 10, 20, 30, and 40% RSM replacing graded levels of SBM, respectively. Fish were fed one of the five experimental diets for eight weeks, and then challenged by A. hydrophila via intraperitoneal injection and kept for seven days. Dietary RSM inclusion level up to 30% did not affect the weight gain and daily growth coefficient, whereas these were depressed by a further inclusion. Fish fed diet with 40% RSM showed the lowest serum total antioxidant capacity, lysozyme, alternative complement pathway, respiratory burst and phagocytic activities. Dietary RSM inclusion gradually depressed the post-challenge survival rate, and that was significantly lower in fish fed diet with 40% RSM compared to fish fed the control diet. Conversely, the inclusion of RSM generally increased the serum total cholesterol level, the plasma alanine aminotransferase and aspartate aminotransferase activities, and these were significantly higher in fish fed diet with 40% RSM compared to fish fed the control diet. The results indicated that RSM can be included at level up to 30% in diet for tilapia without obvious adverse effects on the growth, antioxidant capacity, non-specific immune response and resistance to A. hydrophila infection, whereas these were depressed by a further inclusion.

  19. Optimisation and standardisation of functional immune assays for striped catfish (Pangasianodon hypophthalmus) to compare their immune response to live and heat killed Aeromonas hydrophila as models of infection and vaccination.

    PubMed

    Sirimanapong, Wanna; Thompson, Kim D; Kledmanee, Kan; Thaijongrak, Prawporn; Collet, Bertrand; Ooi, Ei Lin; Adams, Alexandra

    2014-10-01

    Aquaculture production of Pangasianodon hypophthalmus is growing rapidly in South East Asia, especially in Vietnam. As it is a relatively new aquaculture species there are few reports evaluating its immune response to pathogens. Thus, functional assays for P. hypophthalmus were optimised to evaluate both innate and adaptive immune responses, and were then used to examine immune response following stimulation with live and heat-killed Aeromonas hydrophila. These were used as models of infection and vaccination, respectively. Four treatment groups were used, including a control group, a group injected intraperitonally (IP) with adjuvant only, a group injected with heat-killed A. hydrophila (1 × 10(9) cfu ml(-1) mixed with adjuvant), and a group injected with a subclinical dose of live A. hydrophila. Samples were collected at 0, 1, 3, 7, 14 and 21 days post-injection (d.p.i.) to assess their immune response. The results indicated that challenge with live or dead bacteria stimulated the immune response in P. hypophthalmus significantly above the levels observed in control groups with respect to specific antibody titre, plasma lysozyme and peroxidase activity, and phagocytosis by head kidney macrophages at 7 or/and 14 d.p.i. At 21 d.p.i., total and specific antibody (IgM) levels and plasma lysozyme activity in fish injected with either live or dead A. hydrophila were significantly different to the control groups. Differential immune responses were observed between fish injected with either live or dead bacteria, with live A. hydrophila significantly stimulating an increase in WBC counts and plasma peroxidase activity at 3 d.p.i., with the greatest increase in WBC counts noted at 21 d.p.i. and in phagocytosis at 14 d.p.i. By 21 d.p.i. only the macrophages from fish injected with dead A. hydrophila showed significantly stimulation in their respiratory burst activity. This study provides basic information on the immune response in pangasius catfish that can be useful in

  20. Isolation and characterization of Aeromonas species from an eutrophic tropical estuary.

    PubMed

    Marcel, Kouassi Aka; Antoinette, Adingra Ama; Mireille, Dosso

    2002-12-01

    Isolation and characterization of Aeromonas species were undertaken in the Ebrié lagoon (Abidjan, Côte d'Ivoire) over a one year period. Overall 63% of 501 water samples were identified as positive for Aeromonas spp. A. sobria, A. hydrophila and A. caviae represent respectively 49.20%, 20.63% and 30.15% of the positive samples. Thirty nine of the positive samples were recovered from surface water samples; while 24 bottom water samples were positive for Aeromonas. Aeromonas species are most frequently (82.53) present in the Ebrié lagoon urban area during the rainy and flood seasons when the salinity is low and below 10 per thousand. A lower isolation frequency is noticed during the dry season when water salinity is over 10 per thousand. Finally, a higher isolation frequency occurred in samples with highest counts of Escherichia coli.

  1. MONITORING THE EFFECTIVENESS OF UV DISINFECTION OF AEROMONAS SPP. USING SELECTIVE AND NON-SELECTIVE MEDIA

    EPA Science Inventory

    This research was initiated to determine the sensitivity of Aeromonas spp. to ultraviolet (UV) disinfection. Aeromonas hydrophila is a contaminant listed on the USEPA's 1998 CCL. Three different Aeromonas spp. (A. hydrophila, A. sobria and A. caviae) were tested using membrane fi...

  2. Expression of Aeromonas caviae ST pyruvate dehydrogenase complex components mediate tellurite resistance in Escherichia coli

    SciTech Connect

    Castro, Miguel E.; Molina, Roberto C.; Diaz, Waldo A.; Pradenas, Gonzalo A.; Vasquez, Claudio C.

    2009-02-27

    Potassium tellurite (K{sub 2}TeO{sub 3}) is harmful to most organisms and specific mechanisms explaining its toxicity are not well known to date. We previously reported that the lpdA gene product of the tellurite-resistant environmental isolate Aeromonas caviae ST is involved in the reduction of tellurite to elemental tellurium. In this work, we show that expression of A. caviae ST aceE, aceF, and lpdA genes, encoding pyruvate dehydrogenase, dihydrolipoamide transacetylase, and dihydrolipoamide dehydrogenase, respectively, results in tellurite resistance and decreased levels of tellurite-induced superoxide in Escherichia coli. In addition to oxidative damage resulting from tellurite exposure, a metabolic disorder would be simultaneously established in which the pyruvate dehydrogenase complex would represent an intracellular tellurite target. These results allow us to widen our vision regarding the molecular mechanisms involved in bacterial tellurite resistance by correlating tellurite toxicity and key enzymes of aerobic metabolism.

  3. Chitin degradation and metabolism by virulent Aeromonas hydropila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is the causal agent of motile Aeromonas septicemia (MAS) in catfish and other warm-water fishes. Severe outbreak of MAS caused by virulent A. hydrophila (vAh) was reported in 2009 in the Southeastern United States; the disease has since resulted in loss of millions of pounds of ...

  4. Draft genome sequences of three Aeromonas hybrophila isolates from catfish and tilapia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aeromonas hydrophila is a Gram-negative bacteria that is particularly adapted to freshwater environments and can cause severe infections in fish and humans. Here we report the draft genomes of three A. hydrophila catfish and tilapia isolates....

  5. Cloning, Sequencing, and Role in Virulence of Two Phospholipases (A1 and C) from Mesophilic Aeromonas sp. Serogroup O:34

    PubMed Central

    Merino, Susana; Aguilar, Alicia; Nogueras, Maria Mercedes; Regue, Miguel; Swift, Simon; Tomás, Juan M.

    1999-01-01

    Two different representative recombinant clones encoding Aeromonas hydrophila lipases were found upon screening on tributyrin (phospholipase A1) and egg yolk agar (lecithinase-phospholipase C) plates of a cosmid-based genomic library of Aeromonas hydrophila AH-3 (serogroup O34) introduced into Escherichia coli DH5α. Subcloning, nucleotide sequencing, and in vitro-coupled transcription-translation experiments showed that the phospholipase A1 (pla) and C (plc) genes code for an 83-kDa putative lipoprotein and a 65-kDa protein, respectively. Defined insertion mutants of A. hydrophila AH-3 defective in either pla or plc genes were defective in phospholipase A1 and C activities, respectively. Lecithinase (phospholipase C) was shown to be cytotoxic but nonhemolytic or poorly hemolytic. A. hydrophila AH-3 plc mutants showed a more than 10-fold increase in their 50% lethal dose on fish and mice, and complementation of the plc single gene on these mutants abolished this effect, suggesting that Plc protein is a virulence factor in the mesophilic Aeromonas sp. serogroup O:34 infection process. PMID:10417167

  6. Comparison study of membrane filtration direct count and an automated coliform and Escherichia coli detection system for on-site water quality testing.

    PubMed

    Habash, Marc; Johns, Robert

    2009-10-01

    This study compared an automated Escherichia coli and coliform detection system with the membrane filtration direct count technique for water testing. The automated instrument performed equal to or better than the membrane filtration test in analyzing E. coli-spiked samples and blind samples with interference from Proteus vulgaris or Aeromonas hydrophila.

  7. Aeromonas aquariorum Is Widely Distributed in Clinical and Environmental Specimens and Can Be Misidentified as Aeromonas hydrophila▿†

    PubMed Central

    Aravena-Román, Max; Harnett, Gerald B.; Riley, Thomas V.; Inglis, Timothy J. J.; Chang, Barbara J.

    2011-01-01

    Genotypic characterization of 215 Aeromonas strains (143 clinical, 52 environmental, and 20 reference strains) showed that Aeromonas aquariorum (60 strains, 30.4%) was the most frequently isolated species in clinical and water samples and could be misidentified as Aeromonas hydrophila by phenotypic methods. PMID:21697316

  8. Further Characterization of a Type III Secretion System (T3SS) and of a New Effector Protein from a Clinical Isolate of Aeromonas Hydrophila - Part I

    EPA Science Inventory

    A type III secretion system (T3SS)-associated cytotoxin, AexT, with ADP-ribosyltransferase activity and homology to Pseudomonas aeruginosa bifuncational toxins ExoT/S, was recently identified from a fish pathogen Aeromonas salmonicida. In this study, we reported the molecular cha...

  9. ANALYSIS OF AEROMONAS BY MASS SPECTROMETRY: SPECIATION AND VIRULENCE FACTORS

    EPA Science Inventory

    Introduction:

    A number of bacteria, including Aeromonas hydrophila, are listed on the Environmental Protection Agency's 1998 Contaminant Candidate List (CCL) as research needs. One research priority designated by the CCL is the identification of virulence activity facto...

  10. Molecular Characterization and Expression Analyses of the Complement Component C8α, C8β and C9 Genes in Yellow Catfish (Pelteobagrus fulvidraco) after the Aeromonas hydrophila Challenge

    PubMed Central

    Zheng, Huan; Ji, Wei; Zhang, Gui-Rong; Zhang, Xiao-Ting; Shi, Ze-Chao; Wei, Kai-Jian; Yang, Rui-Bin; Gardner, Jonathan P. A.

    2016-01-01

    The complement components C8α, C8β and C9 have important roles in the innate immune system against invading microorganisms. Partial cDNA sequences of the Pf_C8α, Pf_C8β and Pf_C9 genes (Pf: abbreviation of Pelteobagrus fulvidraco) were cloned from yellow catfish. The Pf_C8α, Pf_C8β and Pf_C9 genes showed the greatest amino acid similarity to C8α (54%) and C8β (62%) of zebrafish and to C9 (52%) of grass carp, respectively. Ontogenetic expression analyses using real-time quantitative PCR suggested that the three genes may play crucial roles during embryonic and early larval development. The mRNA expressions of the three genes were all at the highest levels in liver tissue, and at lower or much lower levels in 16 other tissues, demonstrating that the liver is the primary site for the protein synthesis of Pf_C8α, Pf_C8β and Pf_C9. Injection of Aeromonas hydrophila led to up-regulation of the three genes in the spleen, head kidney, kidney, liver and blood tissues, indicating that the three genes may contribute to the host’s defense against invading pathogenic microbes. An increased understanding of the functions of the Pf_C8α, Pf_C8β and Pf_C9 genes in the innate immunity of yellow catfish will help enhance production of this valuable freshwater species. PMID:27005612

  11. Structure of AscE and induced burial regions in AscE and AscG upon formation of the chaperone needle-subunit complex of type III secretion system in Aeromonas hydrophila

    PubMed Central

    Tan, Yih Wan; Yu, Hong Bing; Leung, Ka Yin; Sivaraman, J.; Mok, Yu-Keung

    2008-01-01

    In the type III secretion system (T3SS) of Aeromonas hydrophila, the putative needle complex subunit AscF requires both putative chaperones AscE and AscG for formation of a ternary complex to avoid premature assembly. Here we report the crystal structure of AscE at 2.7 Å resolution and the mapping of buried regions of AscE, AscG, and AscF in the AscEG and AscEFG complexes using limited protease digestion. The dimeric AscE is comprised of two helix–turn–helix monomers packed in an antiparallel fashion. The N-terminal 13 residues of AscE are buried only upon binding with AscG, but this region is found to be nonessential for the interaction. AscE functions as a monomer and can be coexpressed with AscG or with both AscG and AscF to form soluble complexes. The AscE binding region of AscG in the AscEG complex is identified to be within the N-terminal 61 residues of AscG. The exposed C-terminal substrate-binding region of AscG in the AscEG complex is induced to be buried only upon binding to AscF. However, the N-terminal 52 residues of AscF remain exposed even in the ternary AscEFG complex. On the other hand, the 35-residue C-terminal region of AscF in the complex is resistant to protease digestion in the AscEFG complex. Site-directed mutagenesis showed that two C-terminal hydrophobic residues, Ile83 and Leu84, of AscF are essential for chaperone binding. PMID:18662905

  12. Identification of gyrB and rpoB gene mutations and differentially expressed proteins between a novobiocin-resistant Aeromonas hydrophila catfish vaccine strain and its virulent parent strain.

    PubMed

    Pridgeon, J W; Yildirim-Aksoy, M; Klesius, P H; Kojima, K; Mobley, J A; Srivastava, K K; Reddy, P G

    2013-10-25

    A total of 10 and 13 missense mutations were found in the deduced gyrB and rpoB proteins, respectively, between avirulent AH11NOVO vaccine strain and its virulent parent strain AH11P. SDS-PAGE revealed that six proteins bands were significantly over-expressed in AH11NOVO whereas five bands were significantly over-expressed in AH11P. Mass spectrometry identified seven proteins from the over-expressed AH11NOVO gel bands and five proteins from the over-expressed AH11P gel bands. QPCR confirmed that all 12 genes corresponding to the proteins identified by mass spectrometry were significantly over-expressed in AH11NOVO or AH11P. When AH11NOVO proteins were subjected to Western blot analysis, 13 protein bands exhibited significantly stronger reactivity with hyper-immune catfish sera. Fifteen proteins were identified from immunogenic protein bands, including six (formate acetyltransferase, chaperone htpG, transketolase, ATP synthase subunit alpha, asparagine-tRNA ligase, and serine hydroxymethyltransferase) that were over-expressed in AH11NOVO proteins and three (elongation factor G, class II fructose-bisphosphate aldolase, and a putative uncharacterized 23 kDa protein) that were over-expressed in AH11P. In addition, the following six proteins were also identified from the immunogenic protein bands: pyruvate dehydrogenase E1 component, ATP synthase subunit beta, ribose-phosphate pyrophosphokinase, glyceraldehyde-3-phosphate dehydrogenase, 50S ribosomal L10, and 50S ribosomal L15. Our results might provide insights on how to develop novel efficacious vaccine against Aeromonas hydrophila infection.

  13. Molecular Characterization and Expression Analyses of the Complement Component C8α, C8β and C9 Genes in Yellow Catfish (Pelteobagrus fulvidraco) after the Aeromonas hydrophila Challenge.

    PubMed

    Zheng, Huan; Ji, Wei; Zhang, Gui-Rong; Zhang, Xiao-Ting; Shi, Ze-Chao; Wei, Kai-Jian; Yang, Rui-Bin; Gardner, Jonathan P A

    2016-03-08

    The complement components C8α, C8β and C9 have important roles in the innate immune system against invading microorganisms. Partial cDNA sequences of the Pf_C8α, Pf_C8β and Pf_C9 genes (Pf: abbreviation of Pelteobagrus fulvidraco) were cloned from yellow catfish. The Pf_C8α, Pf_C8β and Pf_C9 genes showed the greatest amino acid similarity to C8α (54%) and C8β (62%) of zebrafish and to C9 (52%) of grass carp, respectively. Ontogenetic expression analyses using real-time quantitative PCR suggested that the three genes may play crucial roles during embryonic and early larval development. The mRNA expressions of the three genes were all at the highest levels in liver tissue, and at lower or much lower levels in 16 other tissues, demonstrating that the liver is the primary site for the protein synthesis of Pf_C8α, Pf_C8β and Pf_C9. Injection of Aeromonas hydrophila led to up-regulation of the three genes in the spleen, head kidney, kidney, liver and blood tissues, indicating that the three genes may contribute to the host's defense against invading pathogenic microbes. An increased understanding of the functions of the Pf_C8α, Pf_C8β and Pf_C9 genes in the innate immunity of yellow catfish will help enhance production of this valuable freshwater species.

  14. Molecular cloning, nucleotide sequence, and expression in Escherichia coli of a hemolytic toxin (aerolysin) gene from Aeromonas trota

    SciTech Connect

    Khan, A.A.; Kim, E.; Cerniglia, C.E.

    1998-07-01

    Aeromonas trota AK2, which was derived from ATCC 49659 and produces the extracellular pore-forming hemolytic toxin aerolysin, was mutagenized with the transposon mini-Tn5Km1 to generate a hemolysin-deficient mutant, designated strain AK253. Southern blotting data indicated that an 8.7-kb NotI fragment of the genomic DNA of strain AK253 contained the kanamycin resistance gene of mini-Tn5Km1. The 8.7-kb NotI DNA fragment was cloned into the vector pGEM5Zf({minus}) by selecting for kanamycin resistance, and the resultant clone, pAK71, showed aerolysin activity in Escherichia coli JM109. The nucleotide sequence of the aerA gene, located on the 1.8-kb ApaI-EcoRI fragment, was determined to consist of 1,479 bp and to have an ATG initiation codon and a TAA termination codon. An in vitro coupled transcription-translation analysis of the 1.8-kb region suggested that the aerA gene codes for a 54-kDa protein, in agreement with nucleotide sequence data. The deduced amino acid sequence of the aerA gene product of A. trota exhibited 99% homology with the amino acid sequence of the aerA product of Aeromonas sobria AB3 and 57% homology with the amino acid sequences of the products of the aerA genes of Aeromonas salmonicida 17-2 and A. sobria 33.

  15. Mixture of Sodium Hypochlorite and Hydrogen Peroxide on Adhered Aeromonas hydrophila to Solid Substrate in Water: Impact of Concentration and Assessment of the Synergistic Effect

    PubMed Central

    Lontsi Djimeli, Chrétien; Tamsa Arfao, Antoine; Noah Ewoti, Olive V.; Nougang, Mireille Ebiane; Moungang, Marlyse L.; Nola, Moïse; Sime-Ngando, Télesphore

    2014-01-01

    The synergistic effects of the combined treatments of NaOCl and H2O2 on the elimination of A. hydrophila adhered to polythene under static and dynamic conditions were evaluated. The concentrations 0.1, 0.2, and 0.3‰ NaOCl and 0.5, 1, and 1.5‰ H2O2 were used. The contact periods were 180, 360, 540, and 720 minutes. The abundance of cells adhered reached 2.47 and 2.27 units (log (CFU/cm²)), respectively, under static and dynamic conditions after action of the mixture of disinfectants, whereas it reached 2.41 and 3.39 units (log (CFU/cm²)) after action of NaOCl and H2O2 alone, respectively. Increase in the incubation period resulted in a significant decrease in the abundance of cells adhered when the mixture of 0.3‰ NaOCl and 1.5‰ H2O2 was used (P < 0.01). For each cell growth phase, there was a significant difference amongst the mean densities of cells adhered after action of the mixture of disinfectants (P < 0.05). Although the Freundlich isotherm parameters relatively varied from one experimental condition to another, the Kf value registered in the exponential growth phase was relatively higher in static state than in dynamic regime; cells adhered under dynamic condition seem more sensitive to the synergistic action than those adhered under static condition. PMID:26904729

  16. OVERVIEW: DISINFECTION OF HELICOBACTER PYLORI AND AEROMONAS SPECIES

    EPA Science Inventory

    Helicobacter pylori and Aeromonas hydrophila are contaminants listed on the USEPA's 1998 Contaminant Candidate List (CCL).The sensitivity of H. pylori to chlorine and of Aeromonas spp. to inactivation by free chlorine, chloramine and ultraviolet (UV) was examined. Selective and...

  17. CHARACTERIZATION OF AEROMONAS VIRULENCE USING AN IMMUNOCOMPROMISED MOUSE MODEL

    EPA Science Inventory

    An immunocompromised mouse model was used to characterize Aeromonas strains for their ability to cause opportunistic, extraintestinal infections. A total of 34 isolates of Aeromonas (A. hydrophila [n = 12]), A. veronii biotype sobria [n = 7], A. caviae [n = 4], A. enchelia [n = 4...

  18. Histopathological findings in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with 3 different Aeromonas species.

    PubMed

    Zepeda-Velázquez, Andrea Paloma; Vega-Sánchez, Vicente; Salgado-Miranda, Celene; Soriano-Vargas, Edgardo

    2015-07-01

    This study describes the macroscopic and microscopic lesions in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with genetically identified Aeromonas salmonicida, A. hydrophila, and A. veronii species. The genus Aeromonas includes bacteria that naturally inhabit both waterways and organisms. At least 27 Aeromonas species have been identified to date, some of which can cause significant economic losses in aquaculture. As up to 68.8% of Aeromonas isolates may be misidentified in routine biochemical and phenotypic tests, however, reported cases of Aeromonas infection in fish may be wrongly identified. Our findings confirmed that the 3 Aeromonas species studied are associated with septicemia and dermal lesions in rainbow trout.

  19. Histopathological findings in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with 3 different Aeromonas species

    PubMed Central

    Zepeda-Velázquez, Andrea Paloma; Vega-Sánchez, Vicente; Salgado-Miranda, Celene; Soriano-Vargas, Edgardo

    2015-01-01

    This study describes the macroscopic and microscopic lesions in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with genetically identified Aeromonas salmonicida, A. hydrophila, and A. veronii species. The genus Aeromonas includes bacteria that naturally inhabit both waterways and organisms. At least 27 Aeromonas species have been identified to date, some of which can cause significant economic losses in aquaculture. As up to 68.8% of Aeromonas isolates may be misidentified in routine biochemical and phenotypic tests, however, reported cases of Aeromonas infection in fish may be wrongly identified. Our findings confirmed that the 3 Aeromonas species studied are associated with septicemia and dermal lesions in rainbow trout. PMID:26130859

  20. Aeromonas hydrophila in a burn patient.

    PubMed

    Yasti, Ahmet Cinar; Otan, Emrah; Doganay, Mutlu; Kama, Nuri A

    2009-01-01

    Infectious consequences are still a major problem and leading cause of mortality in burn patients. Among others, aeromonads need special concern because they mimic pseudomonal infections; however, they have a more rapid progression with considerable mortality if undiagnosed promptly. Here, we present a major burn case extinguished with tap water pooled in a tank. With the possibility of aeromonal infection in mind, the patient underwent aggressive debridement with proper antibiotic medication, which resulted in a successful patient management. Aeromonads should always be kept in mind in burn cases that contacted with tanked water or soil after the burn.

  1. Association of Aeromonas sobria with human infection.

    PubMed Central

    Daily, O P; Joseph, S W; Coolbaugh, J C; Walker, R I; Merrell, B R; Rollins, D M; Seidler, R J; Colwell, R R; Lissner, C R

    1981-01-01

    Fifteen Aeromonas isolates from various human infections and nine isolates from polluted water were identified as either Aeromonas hydrophila or Aeromonas sobria and examined for cytotoxigenicity, enterotoxigenicity, adherence to epithelial cells, and other virulence-associated factors, including proteases, lipases, elastases, and hemolysins. Two groups of organisms (I and II) were distinguishable based on differences in median lethal doses in mice and cytotoxicity for Y-1 adrenal cells. Group I clinical and environmental strains had median lethal doses of less than 10(7) colony-forming units, were cytotoxic, frequently possessed several virulence-associated factors, and had lysine decarboxylase-positive or Voges-Proskauer-positive phenotypes or both. Piliation of Aeromonas was associated strongly with ability to adhere to human buccal cells, and these characteristics were associated with group I strains. Group II clinical and environmental strains had median lethal doses of greater than or equal to 10(7) colony-forming units, were not cytotoxic, and usually were lysine decarboxylase negative or Voges-Proskauer negative or both. Clinical strains in group II exhibited enterotoxigenicity, which was not detected in group II environmental strains. A sobria was more frequently associated with human infections; 13 of the 15 clinical strains were A. sobria, and 2 were A. hydrophila. On the other hand, the majority of the environmental strains (seven of nine) were A. hydrophila. Images PMID:6112237

  2. Incidence and virulence characteristics of Aeromonas spp. in fish

    PubMed Central

    Abd-El-Malek, Ashraf M.

    2017-01-01

    Aim: This study was conducted to evaluate the presence of Aeromonas spp. in raw and ready-to-eat (RTE) fish commonly consumed in Assiut city, Egypt, and to determine virulence factors due to they play a key role in their pathogenicity. Materials and Methods: A total of 125 samples of raw and RTE fish samples were taken from different fish markets and fish restaurants in Assiut Governorate and screened for the presence of Aeromonas spp. by enrichment on tryptic soy broth then incubated at 30°C for 24 h. Plating unto the sterile Petri dishes containing Aeromonas agar base to which Aeromonas selective supplement was added. The plates were incubated at 37°C for 24 h. Presumptive Aeromonas colonies were biochemically confirmed and analyzed for pathogenicity by hemolysin production, protease, and lipase detection. Results: The results indicated that raw fish were contaminated with Aeromonas spp. (40% in wild and 36% in cultured Nile tilapia). Regarding RTE, Aeromonas spp. could be isolated with the percentage of 16%, 28% and 20% in fried Bolti, grilled Bolti and fried Bayad, respectively. Out of 35 isolates obtained, 22 were categorized as Aeromonas hydrophila, 12 were classified as Aeromonas sobria and Aeromonas caviae were found in only one isolate. The virulence factors of Aeromonas spp. were detected and the results showed that all isolates produced of hemolysin (91.4%), protease (77.1%), and lipase enzyme (17.1%). Conclusion: This study indicates that the presence of A. hydrophila with virulence potential in fresh and RTE fish may be a major threat to public health. PMID:28246446

  3. Virulence factor-activity relationships (VFAR) with specific emphasis on Aeromonas species (spp.).

    PubMed

    Chopra, Ashok K; Graf, Joerg; Horneman, Amy J; Johnson, Judith A

    2009-01-01

    The human population most commonly inflicted with Aeromonas infection includes young children, the elderly and immunocompromised individuals. Importantly, the isolation rate of Aeromonas species from children suffering from diarrhea is similar in developing and developed countries. It is becoming clear that only a small subset of Aeromonas species belonging to a particular hybridization group causes disease in humans. Human infections with this pathogen occur by consuming contaminated food and water. Aeromonas species were isolated from wounds of patients during the tsunami in southern Thailand. Further, increased numbers of this pathogen were recovered from floodwater samples during Hurricane Katrina in New Orleans. Among various species of Aeromonas, A. hydrophila, A. caviae and A. veronii biovar sobria are mainly responsible for causing disease in humans. Our laboratory has isolated various virulence factors from a diarrheal isolate SSU of A. hydrophila and molecularly characterized them. In addition to various virulence factors produced by Aeromonas species, the status of the immune system plays an important role in inducing disease by this pathogen in the host. Taken together, we have made significant advances in better understanding the pathogenesis of Aeromonas infections, which will help in differentiating pathogenic from non-pathogenic aeromonads. This review covers virulence aspects of a clinical isolate of A. hydrophila.

  4. Clinical significance of Aeromonas species isolated from patients with diarrhea.

    PubMed

    Moyer, N P

    1987-11-01

    A total of 248 strains of Aeromonas spp. were isolated from 3,334 human fecal specimens submitted to a state public health laboratory over a 2-year period to be cultured for enteric pathogens. Cary-Blair transport medium, blood ampicillin agar, and alkaline peptone water enrichment provided optimal recovery of Aeromonas spp. A questionnaire requesting clinical and epidemiological information was sent to physicians, who submitted stool samples for testing, with each laboratory report for 107 consecutive stool isolates of Aeromonas spp. The 56 questionnaires which were completed and returned were analyzed to determine the seasonal distribution of illness and the age and sex distribution of patients; characteristic symptoms; and predisposing factors for gastrointestinal disease caused by Aeromonas spp. It was concluded that some A. hydrophila, A. sobria, and A. caviae strains are capable of causing diarrhea and that antibiotic therapy and the drinking of untreated water are significant risk factors for susceptible hosts.

  5. Clinical significance of Aeromonas species isolated from patients with diarrhea.

    PubMed Central

    Moyer, N P

    1987-01-01

    A total of 248 strains of Aeromonas spp. were isolated from 3,334 human fecal specimens submitted to a state public health laboratory over a 2-year period to be cultured for enteric pathogens. Cary-Blair transport medium, blood ampicillin agar, and alkaline peptone water enrichment provided optimal recovery of Aeromonas spp. A questionnaire requesting clinical and epidemiological information was sent to physicians, who submitted stool samples for testing, with each laboratory report for 107 consecutive stool isolates of Aeromonas spp. The 56 questionnaires which were completed and returned were analyzed to determine the seasonal distribution of illness and the age and sex distribution of patients; characteristic symptoms; and predisposing factors for gastrointestinal disease caused by Aeromonas spp. It was concluded that some A. hydrophila, A. sobria, and A. caviae strains are capable of causing diarrhea and that antibiotic therapy and the drinking of untreated water are significant risk factors for susceptible hosts. PMID:3693537

  6. Analysis of a ferric uptake regulator (Fur) knockout mutant in Aeromonas salmonicida subsp. salmonicida.

    PubMed

    Ebanks, Roger O; Goguen, Michel; Knickle, Leah; Dacanay, Andrew; Leslie, Andrew; Ross, Neil W; Pinto, Devanand M

    2013-03-23

    Aeromonas salmonicida subsp. salmonicida is the etiological agent of furunculosis; a serious infectious disease in aquaculture raised salmonids. Iron acquisition has been shown to be critical for the survival of pathogenic bacteria during the course of infection. Previous work has demonstrated that A. salmonicida expresses iron-repressible IROMP proteins, suggesting the presence of iron acquisition systems that are under the control of a ferric uptake regulator (Fur). In this study, the A. salmonicida fur has been sequenced and a fur deletion strain generated. The A. salmonicida fur gene has an open reading frame of 428 bp, coding for a protein of 143 amino acids, and with high homology to previously described Fur proteins. The Fur protein product had a 94% sequence identity and 96% sequence similarity to the Aeromonas hydrophila Fur protein product. Transcription of the A. salmonicida fur gene was not regulated by the iron status of the bacterium and is not autoregulated, as in Escherichia coli. Proteomic analysis of the A. salmonicida fur mutant, fails to repress iron-regulated outer membrane proteins in the presence of iron. The A. salmonicida fur::KO mutant shows significantly reduced pathogenicity compared to the wild-type parental strain. In addition, the A. salmonicida fur mutant provides an important tool for further investigation of the iron acquisition mechanisms utilized by A. salmonicida.

  7. Prevalence and molecular characterization of Aeromonas spp. in ready-to-eat foods in Italy.

    PubMed

    Villari, P; Crispino, M; Montuori, P; Stanzione, S

    2000-12-01

    A survey was carried out in Italy to ascertain the prevalence of Aeromonas spp. in ready-to-eat foods (vegetables, cheeses, meat products, and ice creams) and the level of molecular heterogeneity of the isolates found by macrorestriction analysis of genomic DNA with pulsed-field gel electrophoresis (PFGE). In total, 46 (14.4%) of the 320 food samples examined were found positive for Aeromonas spp. The highest percentages of isolation were discovered in vegetables, particularly lettuce (45.0%), endive (40.0%), and rucola (20.0%). Ricotta was the only cheese type analyzed that showed a high frequency of isolation (45.0%). Among meat products, salami and raw ham (25.0% of samples positive) and, to a lesser extent, baloney (5.0%) were found positive for Aeromonas spp. Aeromonas hydrophila was the most common isolate from foods of animal origin, whereas Aeromonas caviae was the dominant species in vegetables. No motile aeromonads were found in ice cream samples. Aeromonas isolates showed a high level of genetic heterogeneity, because 24 PFGE patterns were identified among 27 A. hydrophila strains and 20 PFGE patterns were found in 23 A. caviae isolates. In conclusion, consumers of ready-to-eat foods in Italy are regularly exposed to many genetically distinct strains of A. hydrophila and A. caviae without evident signs of malaise, and therefore, few of these strains, if any, are likely to be pathogenic.

  8. Aeromonas primary wound infection of a diver in polluted waters.

    PubMed Central

    Joseph, S W; Daily, O P; Hunt, W S; Seidler, R J; Allen, D A; Colwell, R R

    1979-01-01

    Two separate species of Aeromonas, A. sobria (not listed as a species in Bergey's Manual of Determinative Bacteriology, 8th ed.) and A. hydrophila, were primary pathogens isolated from the leg wound of a diver conducting operations in polluted waters. This is the first recorded instance of a primary infection of soft tissue in a human caused by two species of Aeromonas, one of which was resistant to tetracycline. Because of the very rapid development of this wound infection, cytotoxicity of these organisms was examined in several biological systems. A. sobria was hemolytic for sheep erythrocytes, cytotoxic for Y-1 adrenal cells, and enterotoxic in rabbit ligated intestinal loops, whereas A. hydrophila was hemolytic and cytotoxic. Pertinent clinical, bacteriological, and environmental features of the case are presented. PMID:500794

  9. Clinical and Therapeutic Implications of Aeromonas Bacteremia: 14 Years Nation-Wide Experiences in Korea

    PubMed Central

    Jung, Dong Sik

    2016-01-01

    Background To elucidate the clinical presentation, antimicrobial susceptibility, and prognostic factors of monomicrobial Aeromonas bacteremia in order to determine the most effective optimal therapy. Materials and Methods We reviewed the medical records of Aeromonas bacteremia patients for the period January 2000 to December 2013 in a retrospective multi-center study. Results A total of 336 patient records were reviewed, with 242 having community-acquired bacteremia. The major clinical infections were of the hepatobiliary tract (50.6%) and peritonitis (18.5%), followed by primary bacteremia (17.9%). The infections usually occurred in patients with malignancy (42.3%), hepatic cirrhosis (39.3%), or diabetes mellitus (25.6%). High antimicrobial-resistance rates (15.5% for ceftriaxone, 15.5% for piperacillin/tazobactam) were noted. However, resistance to carbapenem and amikacin was only 9.8% and 3.0%, respectively. Aeromonas hydrophila (58.9%) was the most common pathogen, followed by Aeromonas caviae (30.4%). The severity of A. caviae bacteremia cases were less than that of A. hydrophila or Aeromonas veronii bacteremia (P <0.05). A. hydrophila showed higher antimicrobial resistance than did other Aeromonas species (P <0.05). Patients with hospital-acquired bacteremia were more likely to have severely abnormal laboratory findings and relatively high antimicrobial-resistance rates. Mortality was associated with metastatic cancer, shock, delayed use of appropriate antimicrobial agents, increased prothrombin time, and increased creatinine level (P <0.05). Conclusions Aeromonas species should be considered one of the causative agents of bacteremia in patients with intra-abdominal infections or malignancies. Although ceftriaxone-resistant Aeromonas bacteremia was not statistically related to mortality in this study, it was associated with severe clinical manifestations and laboratory abnormalities. Appropriate antibiotics, including carbapenem, should be administered early

  10. Reassessment of the Enteropathogenicity of Mesophilic Aeromonas Species

    PubMed Central

    Teunis, Peter; Figueras, Maria J.

    2016-01-01

    Cases of Aeromonas diarrhea have been described all over the world. The genus Aeromonas includes ca. 30 species, of which 10 have been isolated in association with gastroenteritis. The dominating species that account for ca. 96% of the identified strains are Aeromonas caviae, A. veronii, A. dhakensis, and A. hydrophila. However, the role of Aeromonas as a true enteropathogen has been questioned on the basis of the lack of outbreaks, the non-fulfillment of Koch’s postulates and the low numbers of acute illnesses in the only existing human challenge study. In the present study we reassess the enteropathogenicity of Aeromonas using dose response models for microbial infection and acute illness. The analysis uses the data from the human challenge study and additional data from selected outbreak investigations where the numbers exposed and the dose were reported, allowing their inclusion as “natural experiments”. In the challenge study several cases of asymptomatic shedding were found (26.3%, 15/57), however, only 3.5% (2/57) of those challenged with Aeromonas developed acute enteric symptoms (i.e., diarrhea). The “natural experiments” showed a much higher risk of illness associated with exposure to Aeromonas, even at moderate to low doses. The median dose required for 1% illness risk, was ~1.4 × 104 times higher in the challenge study (1.24 × 104 cfu) compared to natural exposure events (0.9 cfu). The dose response assessment presented in this study shows that the combined challenge and outbreak data are consistent with high infectivity of Aeromonas, and a wide range of susceptibility to acute enteric illness. To illustrate the outcomes, we simulate the risk associated with concentrations of Aeromonas found in different water and food matrices, indicating the disease burden potentially associated with these bacteria. In conclusion this study showed that Aeromonas is highly infectious, and that human susceptibility to illness may be high, similar to

  11. Molecular Characterization of a Functional Type VI Secretion System from a Clinical Isolate of Aeromonas hydrophilia

    EPA Science Inventory

    Our laboratory recently molecularly characterized the type II secretion system (T2SS)-associated cytotoxic enterotoxin (Act) and the T3SS-secreted AexU effector from a diarrheal isolate SSU of Aeromonas hydrophila. The role of these toxin proteins in the pathogenesis of A. hydrop...

  12. VIRULENCE RELATIONSHIPS OF AEROMONAS SPECIES AS DETERMINED BY EXPOSURES TO IMMUNOCOMPROMISED MICE

    EPA Science Inventory

    Our laboratory is currently determining the virulence of opportunistic pathogens reported in treated drinking water and drinking water sources. Aeromonas hydrophila is currently on the EPA's Contaminant Candidate List (CCL) and is an example of those types of bacteria that conta...

  13. HOST GENE CELL RESEARCH FOR DETERMINING VIRULENCE OF AEROMONAS SPP. COLLECTED FROM ENVIRONMENTAL SAMPLES

    EPA Science Inventory

    The United States Environmental Protection Agency (USEPA) is interested in assessing health risks associated with emerging or potential waterborne pathogens. To this end, the Agency has established a Candidate Contaminant List (CCL) that includes Aeromonas hydrophila an...

  14. Tetrahymena: An Alternative Model Host for Evaluating Virulence of Aeromonas Strains

    PubMed Central

    Pang, Mao-Da; Lin, Xiao-Qin; Hu, Meng; Li, Jing; Lu, Cheng-Ping; Liu, Yong-Jie

    2012-01-01

    An easier assessment model would be helpful for high-throughput screening of Aeromonas virulence. The previous study indicated the potential of Tetrahymena as a permissive model to examine virulence of Aeromonas hydrophila. Here our aim was to assess virulence of Aeromonas spp. using two model hosts, a zebrafish assay and Tetrahymena-Aeromonas co-culture, and to examine whether data from the Tetrahymena thermophila model reflects infections in the well-established animal model. First, virulence of 39 Aeromonas strains was assessed by determining the 50% lethal dose (LD50) in zebrafish. LD50 values ranging from 1.3×102 to 3.0×107 indicated that these strains represent a high to moderate degree of virulence and could be useful to assess virulence in the Tetrahymena model. In Tetrahymena-Aeromonas co-culture, we evaluated the virulence of Aeromonas by detecting relative survival of Aeromonas and Tetrahymena. An Aeromonas isolate was considered virulent when its relative survival was greater than 60%, while the Aeromonas isolate was considered avirulent if its relative survival was below 40%. When relative survival of T. thermophila was lower than 40% after co-culture with an Aeromonas isolate, the bacterial strain was regarded as virulent. In contrast, the strain was classified as avirulent if relative survival of T. thermophila was greater than 50%. Encouragingly, data from the 39 Aeromonas strains showed good correlation in zebrafish and Tetrahymena-Aeromonas co-culture models. The results provide sufficient data to demonstrate that Tetrahymena can be a comparable alternative to zebrafish for determining the virulence of Aeromonas isolates. PMID:23145022

  15. Development of an Aeromonas hydrophila  infection model using the protozoan Tetrahymena thermophila.

    PubMed

    Li, Jing; Zhang, Xiao-Lu; Liu, Yong-Jie; Lu, Cheng-Ping

    2011-03-01

    Aeromonas hydrophila is a motile bacterium present in numerous freshwater habitats worldwide and is frequently the cause of infections in fish and numerous terrestrial vertebrates including humans. Because A. hydrophila is also a component of the normal intestinal flora of healthy fish, virulence mechanisms are not well understood. Considering that fish models used for the examination of A. hydrophila genes associated with virulence have not been well defined, we established an infection model using the free-living, ciliate protozoa Tetrahymena thermophila. The expression of A. hydrophila virulence genes following infection of T. thermophila was assessed by reverse transcription-PCR and demonstrated that the aerolysin (aerA) and Ahe2 serine protease (ahe2) genes (not present in the avirulent A. hydrophila NJ-4 strain) in the virulent J-1 strain were upregulated 4-h postinfection. Furthermore, the presence of intact A. hydrophila J-1 within T. thermophila suggested that these bacteria could interfere with phagocytosis, resulting in the death of the infected protozoan 48-h postinfection. Conversely, A. hydrophila NJ-4-infected T. thermophila survived the infection. This study established a novel T. thermophila infection model that will provide a novel means of examining virulence mechanisms of A. hydrophila.

  16. Effectiveness of radiation processing in elimination of Aeromonas from food

    NASA Astrophysics Data System (ADS)

    Nagar, Vandan; Bandekar, Jayant R.

    2011-08-01

    Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D10) values of different Aeromonas isolates in saline at 0-4 °C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839T and A. veronii Y47) and exposed to γ radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D10 values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081±0.001, 0.089±0.003 and 0.091±0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 105 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 °C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 105 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

  17. Isolation and characterization of Aeromonas from seafoods in Taipei.

    PubMed

    Yaun, S S; Lin, L P

    1993-05-01

    A total of 124 fresh seafoods and 158 processed seafoods collected from the retail markets and supermarkets in Taipei were tested for the contamination with motile Aeromonas spp. Of the fresh seafoods analyzed, 88% displayed the presence of Aeromonas. The isolation rates of various samples were as follows: 100%, freshwater fish; 95%, seawater fish; 78%, fish fillets; 84%, shrimp and crab of the crustacea group; 83%, bivalve shellfish and 84%, non-bivalve shellfish of the mollusca group, and 100%, seaweed. Of the 158 processed seafoods, 11% were contaminated by Aeromonas. The isolation rates were as follows: 0%, canned, dried, or frozen fresh seafood; 18%, salted seafood; 30%, fish cake; 7% vacuum-packaged fish cakes; 14%, frozen seafood dumplings; 8%, cooked seafoods. One hundred and eighty-three Aeromonas strains isolated in this survey were characterized to species level and tested for their ability to produce beta-hemolysin. Ninety-eight percent (98%) of the A. hydrophila produced beta-hemolysin on 5% blood agar, 94% of the A. sobria and 33% of the A. caviae produced beta-hemolysin. Thus it is likely that fresh seafoods are potentially significant sources of the virulent Aeromonas species and may play an important role in the epidemiology of Aeromonas-associated gastroenteritis.

  18. Characterization of Aeromonas species isolated from an estuarine environment

    PubMed Central

    Evangelista-Barreto, Norma Suely; de Carvalho, Fatima Cristiane Teles; Vieira, Regine Helena S. dos Fernandes; dos Reis, Cristhiane Moura Falavina; Macrae, Andrew; Rodrigues, Dália dos Prazeres

    2010-01-01

    Thirty water samples were collected, at two week intervals, from the estuary of the River Cocó. The aim was to characterize the presence, distribution and types of Aeromonas spp, in the estuary of the River Cocó, Ceara, Brazil (03°46’28.83’’S e 38°26’36.52’’S). Aeromonas were identified in 19 (63%) samples analyzed by plating and CFU counts. Presence/absence tests were positive for 11 (37%) of the samples resulting in the detection of Aeromonas in a total of 23 (77%) of samples. CFU counts varied from < 10 to 1.4 x 104 CFU mL-1. From the isolated strains seven species of Aeromonas were identified: A. caviae (29/69), A. veronii bv. sobria (13/69), A. veronii bv. veronii (8/69), A. trota (6/69), A. media (5/69), A. sobria (4/69) and A. hydrophila and Aeromonas sp. (2/69). Of the 38 strains tested, 23 (60%) showed resistance to at least one of the eight antimicrobials. Multiple resistance to antibiotics was observed in A. caviae, A. media , A. sóbria and A. veronii bv. sobria. Aeromonas caviae showed the highest multiple resistance, being resistant to four antibiotics. The presence of those microorganisms may contribute to the occurrence of gastroenteritis, mainly in children, since they are considered opportunists. PMID:24031516

  19. The Genus Aeromonas: Taxonomy, Pathogenicity, and Infection

    PubMed Central

    Janda, J. Michael; Abbott, Sharon L.

    2010-01-01

    Summary: Over the past decade, the genus Aeromonas has undergone a number of significant changes of practical importance to clinical microbiologists and scientists alike. In parallel with the molecular revolution in microbiology, several new species have been identified on a phylogenetic basis, and the genome of the type species, A. hydrophila ATCC 7966, has been sequenced. In addition to established disease associations, Aeromonas has been shown to be a significant cause of infections associated with natural disasters (hurricanes, tsunamis, and earthquakes) and has been linked to emerging or new illnesses, including near-drowning events, prostatitis, and hemolytic-uremic syndrome. Despite these achievements, issues still remain regarding the role that Aeromonas plays in bacterial gastroenteritis, the extent to which species identification should be attempted in the clinical laboratory, and laboratory reporting of test results from contaminated body sites containing aeromonads. This article provides an extensive review of these topics, in addition to others, such as taxonomic issues, microbial pathogenicity, and antimicrobial resistance markers. PMID:20065325

  20. CHARACTERIZATION OF PRECURSOR 165 RRNA FOR AEROMONAS HYDROPHILA

    EPA Science Inventory

    Current strategies for monitoring drinking water quality involve culture-based methods to detect the presence of microbial indicators. However, these methods are insensitive when the organisms have undergone physiological changes such as injury and starvation that can occur in h...

  1. Influence of food system conditions on N-acyl-L-homoserine lactones production by Aeromonas spp.

    PubMed

    Medina-Martínez, M S; Uyttendaele, M; Demolder, V; Debevere, J

    2006-12-01

    Eleven of 13 Aeromonas strains were shown to produce AHLs. Results of TLC showed that N-butanoyl-L-homoserine lactone (C4-HSL) was the main AHL produced in LB medium at 30 degrees C. The influence of different carbon sources, temperature, pH values and salt concentrations on AHL production was determined in eight A. hydrophila and one A. caviae strain. Additionally a quantitative study of C4-HSL production by A. hydrophila strain 519 under different conditions was performed. Positive results were found in the AHL induction assay for some Aeromonas strains in cultures in LB agar incubated at 12 degrees C after 72-96 h. The induction of the sensor strains by Aeromonas spp. occurred in LB medium supplemented with all carbon sources in a concentration of 0.5%. The production of C4-HSL by A. hydrophila 519 was found until 3.5% (w/v) of NaCl. For pHs close to the neutrality the C4-HSL production by A. hydrophila was evident after 24-48 h of incubation. A. hydrophila 519 produced C4-HSL under anaerobic conditions. Also, the AHL production by Aeromonas strains was studied in simulate agar of shrimp, fish and some vegetables. The production of AHLs was evident by almost all the test strains in shrimp simulated agar. In fish agar only for one of three fish species tested, positive results were found. Induction assay in vegetables simulated agar showed principally negative results, probably because of the presence of inhibitory compounds in these vegetables.

  2. Antimicrobial susceptibility testing of aquatic bacteria: quality control disk diffusion ranges for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28 degrees C.

    PubMed

    Miller, R A; Walker, R D; Baya, A; Clemens, K; Coles, M; Hawke, J P; Henricson, B E; Hsu, H M; Mathers, J J; Oaks, J L; Papapetropoulou, M; Reimschuessel, R

    2003-09-01

    Quality control (QC) ranges for disk diffusion susceptibility testing of aquatic bacterial isolates were proposed as a result of a multilaboratory study conducted according to procedures established by the National Committee for Clinical Laboratory Standards (NCCLS). Ranges were proposed for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28 degrees C for nine different antimicrobial agents (ampicillin, enrofloxacin, erythromycin, florfenicol, gentamicin, oxolinic acid, oxytetracycline, ormetoprim-sulfadimethoxine, and trimethoprim-sulfamethoxazole). All tests were conducted on standard Mueller-Hinton agar. With >/=95% of all data points fitting within the proposed QC ranges, the results from this study comply with NCCLS guidelines and have been accepted by the NCCLS Subcommittee for Veterinary Antimicrobial Susceptibility Testing. These QC guidelines will permit greater accuracy in interpreting results and, for the first time, the ability to reliably compare susceptibility test data between aquatic animal disease diagnostic laboratories.

  3. Antimicrobial Susceptibility Testing of Aquatic Bacteria: Quality Control Disk Diffusion Ranges for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28°C

    PubMed Central

    Miller, R. A.; Walker, R. D.; Baya, A.; Clemens, K.; Coles, M.; Hawke, J. P.; Henricson, B. E.; Hsu, H. M.; Mathers, J. J.; Oaks, J. L.; Papapetropoulou, M.; Reimschuessel, R.

    2003-01-01

    Quality control (QC) ranges for disk diffusion susceptibility testing of aquatic bacterial isolates were proposed as a result of a multilaboratory study conducted according to procedures established by the National Committee for Clinical Laboratory Standards (NCCLS). Ranges were proposed for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 at 22 and 28°C for nine different antimicrobial agents (ampicillin, enrofloxacin, erythromycin, florfenicol, gentamicin, oxolinic acid, oxytetracycline, ormetoprim-sulfadimethoxine, and trimethoprim-sulfamethoxazole). All tests were conducted on standard Mueller-Hinton agar. With ≥95% of all data points fitting within the proposed QC ranges, the results from this study comply with NCCLS guidelines and have been accepted by the NCCLS Subcommittee for Veterinary Antimicrobial Susceptibility Testing. These QC guidelines will permit greater accuracy in interpreting results and, for the first time, the ability to reliably compare susceptibility test data between aquatic animal disease diagnostic laboratories. PMID:12958263

  4. Pathogenicity of Mexican isolates of Aeromonas sp. in immersion experimentally-infected rainbow trout (Oncorhynchus mykiss, Walbaum 1792).

    PubMed

    Zepeda-Velázquez, Andrea Paloma; Vega-Sánchez, Vicente; Ortega-Santana, César; Rubio-Godoy, Miguel; de Oca-Mira, Dany Montes; Soriano-Vargas, Edgardo

    2017-05-01

    Ten species of Aeromonas have been previously identified in farmed rainbow trout from Mexico. The aim of the current study was to investigate the pathogenicity of 10 Aeromonas isolates belonging to 10 different Aeromonas species in immersion experimentally-infected rainbow trout fry. Isolates of A. bestiarum, A. hydrophila, A. salmonicida, and A. veronii produced significant mortality (8.8%, 12.2%, 18.8%, and 8.8%, respectively). Isolates of A. caviae and A. sobria produced no significantly mortality (3.3% and 1.1%, respectively). No mortality was recorded in fish infected with A. allosaccharophila, A. lusitana, A. media, or A. popoffii. Microscopic lesions and bacterial reisolation were registered in liver of fish infected with the ten different Aeromonas isolates. Our results suggest that all Aeromonas species included in the study have the ability to colonize the liver. The results have confirmed that species A. bestiarum, A. hydrophila, A. salmonicida, and A. veronii affected fish as elsewhere reported. In conclusion, the variation in pathogenicity of Aeromonas isolates included in the study, emphasizes the importance of active, on-going monitoring of Aeromonas in the Mexican rainbow trout farming.

  5. Aeromonas dhakensis, an Increasingly Recognized Human Pathogen

    PubMed Central

    Chen, Po-Lin; Lamy, Brigitte; Ko, Wen-Chien

    2016-01-01

    Aeromonas dhakensis was first isolated from children with diarrhea in Dhaka, Bangladesh and described in 2002. In the past decade, increasing evidence indicate this species is widely distributed in the environment and can cause a variety of infections both in human and animals, especially in coastal areas. A. dhakensis is often misidentified as A. hydrophila, A. veronii, or A. caviae by commercial phenotypic tests in the clinical laboratory. Correct identification relies on molecular methods. Increasingly used matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) may be able to identify Aeromonas specie rapidly and accurately. A. dhakensis has shown its potent virulence in different animal models and clinical infections. Although several virulence factors had been reported, no single mechanism is conclusive. Characteristically A. dhakensis is the principal species causing soft tissue infection and bacteremia, especially among patients with liver cirrhosis or malignancy. Of note, A. dhakensis bacteremia is more lethal than bacteremia due to other Aeromonas species. The role of this species in gastroenteritis remains controversial. Third generation cephalosporins and carbapenems should be used cautiously in the treatment of severe A. dhakensis infection due to the presence of AmpC ββ-lactamase and metallo-β-lactamase genes, and optimal regimens may be cefepime or fluoroquinolones. Studies of bacterial virulence factors and associated host responses may provide the chance to understand the heterogeneous virulence between species. The hypothesis A. dhakensis with varied geographic prevalence and enhanced virulence that compared to other Aeromonas species warrants more investigations. PMID:27303382

  6. Isolation of Aeromonas spp. from an unchlorinated domestic water supply.

    PubMed Central

    Burke, V; Robinson, J; Gracey, M; Peterson, D; Meyer, N; Haley, V

    1984-01-01

    The recovery of Aeromonas spp. from the unchlorinated water supply for a Western Australian city of 21,000 people was monitored at several sampling points during a period of 1 year. Membrane filtration techniques were used to count colonies of Aeromonas spp., coliforms, and Escherichia coli in water sampled before entry to service reservoirs, during storage in service reservoirs, and in distribution systems. Aeromonas spp. were identified by subculture on blood agar with ampicillin, oxidase tests, and the use of Kaper medium and then were tested for production of enterotoxins and hemolysins. During the same period, two-thirds of all fecal specimens sent for microbiological examination were cultured on ampicillin-blood agar for Aeromonas spp. Recovery of Aeromonas spp. from water supplies at distribution points correlated with fecal isolations and continued during autumn and winter. Coliforms and E. coli were found most commonly in late summer to autumn. This pattern differs from the summer peak of Aeromonas isolations both from water and from patients with Aeromonas spp.-associated gastroenteritis in Perth, Western Australia, a city with a chlorinated domestic water supply. Of the Aeromonas strains from water, 61% were enterotoxigenic, and 64% produced hemolysins. PMID:6486783

  7. Faecal contamination indicators, Salmonella, Vibrio and Aeromonas in water used for the irrigation of agricultural products.

    PubMed Central

    Pianietti, A.; Sabatini, L.; Bruscolini, F.; Chiaverini, F.; Cecchetti, G.

    2004-01-01

    The faecal contamination indicators (total coliforms, faecal coliforms, Escherichia coli, enterococci) and the genera Salmonella, Vibrio, Aeromonas were investigated in water samples used for irrigation. During 4 months, 52 samples were taken. The methods used were: multiple tube fermentation method for faecal contamination indicators and membrane filtration techniques for salmonella, aeromonas and vibrio. Two samples were positive for Salmonella spp., fourteen for Aeromonas spp. and no samples for Vibrio spp. No correlation was found between aeromonas and the indicators of faecal contamination. Regarding Aeromonas spp., 21.6% of the strains were adhesive and 12.6% cytotoxic: this confirms the possible role of aeromonas in human pathologies. These results are important to determine the quality of irrigation water in relation to human health. In fact, the spray or sprinkler irrigation produces bioaerosol, which can contaminate the crops that are likely to be eaten uncooked. In addition, the flood or furrow irrigation represents a risk to field workers. PMID:15061497

  8. Aeromonas and Plesiomonas species from scarlet ibis (Eudocimus ruber) and their environment: monitoring antimicrobial susceptibility and virulence.

    PubMed

    Castelo-Branco, Débora Souza Collares Maia; Silva, Aline Lobão da; Monteiro, Frederico Ozanan Barros; Guedes, Glaucia Morgana de Melo; Sales, Jamille Alencar; Oliveira, Jonathas Sales de; Maia Junior, José Erisvaldo; Miranda, Stefânia Araújo; Sidrim, José Júlio Costa; Alencar, Lucas Pereira de; Brilhante, Raimunda Sâmia Nogueira; Cordeiro, Rossana de Aguiar; Bandeira, Tereza de Jesus Pinheiro Gomes; Pereira Neto, Waldemiro de Aquino; Rocha, Marcos Fábio Gadelha

    2017-01-01

    The present study aimed at evaluating the role of captive scarlet ibises (Eudocimus ruber) and their environment as reservoirs of Aeromonas spp. and Plesiomonas spp., and analyzing the in vitro antimicrobial susceptibility and virulence of the recovered bacterial isolates. Thus, non-lactose and weak-lactose fermenting, oxidase positive Gram-negative bacilli were recovered from cloacal samples (n = 30) of scarlet ibises kept in a conservational facility and from water samples (n = 30) from their environment. Then, the antimicrobial susceptibility, hemolytic activity and biofilm production of the recovered Aeromonas spp. and Plesiomonas shigelloides strains were assessed. In addition, the virulence-associated genes of Aeromonas spp. were detected. Ten Aeromonas veronii bv. sobria, 2 Aeromonas hydrophila complex and 10 P. shigelloides were recovered. Intermediate susceptibility to piperacillin-tazobactam and cefepime was observed in 2 Aeromonas spp. and 1 P. shigelloides, respectively, and resistance to gentamicin was observed in 4 P. shigelloides. The automated susceptibility analysis revealed resistance to piperacillin-tazobactam and meropenem among Aeromonas spp. and intermediate susceptibility to gentamicin among P. shigelloides. All Aeromonas isolates presented hemolytic activity, while 3 P. shigelloides were non-hemolytic. All Aeromonas spp. and 3/10 P. shigelloides were biofilm-producers, at 28 °C, while 10 Aeromonas spp. and 6/10 P. shigelloides produced biofilms, at 37 °C. The most prevalent virulence genes of Aeromonas spp. were asa1 and ascV. Scarlet ibises and their environment harbour potentially pathogenic bacteria, thus requiring monitoring and measures to prevent contamination of humans and other animals.

  9. Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.

    PubMed

    Khor, Wei Ching; Puah, Suat Moi; Tan, Jin Ai Mary Anne; Puthucheary, S D; Chua, Kek Heng

    2015-01-01

    Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

  10. Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia

    PubMed Central

    Khor, Wei Ching; Puah, Suat Moi; Tan, Jin Ai Mary Anne; Puthucheary, SD; Chua, Kek Heng

    2015-01-01

    Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions—exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism. PMID:26710336

  11. Isolation, Enumeration, and Characterization of Aeromonas from Polluted Waters Encountered in Diving Operations

    PubMed Central

    Seidler, Ramon J.; Allen, D. A.; Lockman, H.; Colwell, R. R.; Joseph, S. W.; Daily, O. P.

    1980-01-01

    Counts of total viable, aerobic, heterotrophic bacteria, indicator organisms, and Aeromonas spp. were made at a diver training site on the Anacostia River in Washington, D.C. The numbers of Aeromonas cells in Anacostia River sediment and water increased during periods of elevated water temperature, to maxima of 4 × 105 cells per g of sediment and 300 cells per ml of water. Correspondingly, Aeromonas counts dropped 2 to 4 logs as the water temperature decreased to 0 to 0.5°C. Cultures taken by sterile swabs from the ears and face masks of divers after a 30-min swim in the Anacostia River yielded bacterial types and numbers similar to those found in the river. The nasal passages of the divers apparently did not become contaminated by swimming, possibly because of the protective effect of the face masks used by the divers. Properties associated with virulence in Aeromonas hydrophila and Aeromonas sobria strains isolated from the river, sediment, and divers were investigated. Nearly 40% of the strains of both species collected during the study produced cytotoxic activity for mouse Y-1 adrenal cells, as well as elastase. Enterotoxin activity, as detected by the Y-1 assay, was observed in 3% (1 of 35) of the strains of A. sobria and in 6% (19 of 330) of the A. hydrophila strains. Fluid accumulation in rabbit ileal loops induced by both species of Aeromonas varied greatly among the 17 strains examined. Fluid accumulation of at least 0.4 ml/cm was correlated with positive cytotoxin- or enterotoxin-like response in the Y-1 tissue culture assay. PMID:7396482

  12. Isolation, enumeration, and characterization of Aeromonas from polluted waters encountered in diving operations.

    PubMed

    Seidler, R J; Allen, D A; Lockman, H; Colwell, R R; Joseph, S W; Daily, O P

    1980-05-01

    Counts of total viable, aerobic, heterotrophic bacteria, indicator organisms, and Aeromonas spp. were made at a diver training site on the Anacostia River in Washington, D.C. The numbers of Aeromonas cells in Anacostia River sediment and water increased during periods of elevated water temperature, to maxima of 4 x 10(5) cells per g of sediment and 300 cells per ml of water. Correspondingly, Aeromonas counts dropped 2 to 4 logs as the water temperature decreased to 0 to 0.5 degrees C. Cultures taken by sterile swabs from the ears and face masks of divers after a 30-min swim in the Anacostia River yielded bacterial types and numbers similar to those found in the river. The nasal passages of the divers apparently did not become contaminated by swimming, possibly because of the protective effect of the face masks used by the divers. Properties associated with virulence in Aeromonas hydrophila and Aeromonas sobria strains isolated from the river, sediment, and divers were investigated. Nearly 40% of the strains of both species collected during the study produced cytotoxic activity for mouse Y-1 adrenal cells, as well as elastase. Enterotoxin activity, as detected by the Y-1 assay, was observed in 3% (1 of 35) of the strains of A. sobria and in 6% (19 of 330) of the A. hydrophila strains. Fluid accumulation in rabbit ileal loops induced by both species of Aeromonas varied greatly among the 17 strains examined. Fluid accumulation of at least 0.4 ml/cm was correlated with positive cytotoxin- or enterotoxin-like response in the Y-1 tissue culture assay.

  13. Virulence properties and enterotoxin production of Aeromonas strains isolated from fish.

    PubMed Central

    Santos, Y; Toranzo, A E; Barja, J L; Nieto, T P; Villa, T G

    1988-01-01

    The biological activities in vivo and in vitro of 59 motile Aeromonas spp. isolated from fish and water tanks were simultaneously analyzed in poikilothermic and homoiothermic systems. A total of 64.3% of the isolates tested were pathogenic for fish, and 62% of Aeromonas hydrophila and A. sobria isolates either virulent or nonvirulent for fish were enterotoxigenic. Although the majority of the strains were proteolytic and amylolytic and produced DNase, other activities, such as elastase and staphylolysis, were only present in A. hydrophila. Most of the strains (96%) produced hemolysins, and 68% had agglutinating capacity, but neither isolates pathogenic for fish nor enterotoxigenic isolates showed specificity for trout or human erythrocytes, respectively. The production of siderophores, agglutination in acriflavine, and precipitation after boiling were found not to be useful tests for screening virulent strains. Although statistical analysis revealed a significant relationship between virulence for fish and positive results for arabinose and sucrose fermentations, elastase, and hemolysis of human erythrocytes, only lysine decarboxylase showed a significant positive relationship with enterotoxigenicity. Using extracellular products from representative Aeromonas strains with different virulence markers and belonging to distinct O serogroups, we demonstrated a lack of correlation between cytotoxicity for fish and homoiothermic cell lines and pathogenicity. The extracellular products from selected pathogenic A. hydrophila strains were lethal for rainbow trout and displayed proteolytic, hemolytic, and cytotoxic activities which were simultaneously lost after heat treatment. The findings reported here indicate that it is not possible to establish a common and single mechanism involved in the invasion of Aeromonas spp. in poikilothermic and homoiothermic hosts. Images PMID:2972627

  14. CHLORINE DISINFECTION OF AEROMONAS

    EPA Science Inventory

    The bacterial genus Aeromonas is currently listed on the USEPA's Candidate Contaminant List (CCL). Resistance to chemical disinfection is an essential aspect regarding all microbial groups listed on the CCL. This study was designed to determine the inactivation kinetics of Aeromo...

  15. Aeromonas presence in drinking water from collective reservoirs and wells in peri-urban area in Brazil.

    PubMed

    Pepe Razzolini, Maria Tereza; Risso Günther, Wanda Maria; Martone-Rocha, Solange; Duarte de Luca, Heloísa; Alves Cardoso, Maria Regina

    2010-07-01

    Aeromonas genus is considered an emerging pathogen and its presence in drinking water supplies is a reason to public health concern. This study investigated the occurrence of Aeromonas in samples from collective reservoirs and wells used as drinking water sources in a peri-urban area. A total of 35 water samples were collected from collective reservoirs and 32 from wells bimonthly, from September 2007 to September 2008. Aeromonas spp determination was carried out using a Multiple-Tube Technique. Samples were inoculated into alkaline peptone water and the superficial film formed was transferred to blood agar plates amended with ampicillin. Typical Aeromonas colonies were submitted to a biochemical screening and then to biochemical tests for species differentiation. Aeromonas was detected in 13 (19%) of the 69 samples examined (6 from collective reservoirs and 7 from wells). Concentrations of Aeromonas in collective reservoirs ranged from <0.3 to 1.2 x10(2)MPN/100mL and, in wells, from <0.3 to 2.4 x10(2)MPN/100mL. The most frequent specie in the collective reservoir samples was Aeromonas spp (68%), followed by A. encheleia (14%) and A. allosaccharophila (8%) and A. hydrophila (8%). Aeromonas spp (87%) was the most frequent specie isolated from well samples, followed by A. allosacchariphila (8%), A. encheleia (2%) and A. jandaei (5%). These data show the presence and diversity of Aeromonas genus in the samples analyzed and highlight that its presence in drinking water poses a significant public health concern.

  16. Reduction of Aeromonas hidrophyla biofilm on stainless stell surface by essential oils

    PubMed Central

    Millezi, Alessandra Farias; Cardoso, Maria das Graças; Alves, Eduardo; Piccoli, Roberta Hilsdorf

    2013-01-01

    This study demonstrates the possibility of using sanitizing detergents based on natural products for the elimination and/or reduction of Aeromonas hydrophila biofilm formed on stainless steel surfaces. The goal of this work was to determine the reduction effect of sanitizing detergents containing essential oils of Thymus vulgaris (thyme) and Cymbopogon citratus (lemongrass) on biofilm formed by A. hydrophila on AISI 304 stainless steel coupons, using UHT skimmed milk as substratum. There was adhesion and biofilm formation by A. hydrophila at 28 °C, presenting 7.60 log cfu.cm−2 after the fourth day of cultivation. There was no significant difference between the lemongrass treatment and that of the thyme oil (p < 0.05). However, both treatments significantly reduced the biofilm, differing significantly from the NaOH control (p > 0.05). The treatment with lemongrass solution reduced the biofilm by 4.51 log cfu cm−2 at 25 °C. The thyme detergent also reduced the number of cfu cm−2 by 3.84 log cycles at 25 °C. The use of the lemongrass and thyme solutions efficiently reduced the A. hydrophila biofilm. PMID:24159286

  17. Incidence of Aeromonas spp. infection in fish and chicken meat and its related public health hazards: A review

    PubMed Central

    Praveen, Praveen Kumar; Debnath, Chanchal; Shekhar, Shashank; Dalai, Nirupama; Ganguly, Subha

    2016-01-01

    Aeromonas is recognized to cause a variety of diseases in man. In humans, they are associated with intestinal and extra-intestinal infections. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors such as enterotoxins, hemolysins or cytotoxins, and antibiotic resistance against different antibiotics. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. Comprehensive enteric disease surveillance strategies, prevention and education are essential for meeting the challenges in the years ahead. It is important for us to promote the value of enteric cultures when patients have a gastrointestinal illness or bloody diarrhea or when multiple cases of enteric disease occur after a common exposure. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors, such as enterotoxins, hemolysins or cytotoxins. It has been established that aerolysin is a virulence factor contributing to the pathogenesis of Aeromonas hydrophila infection. Fish and chicken play an important role in the transmission of this pathogen to humans. In the present study, the high prevalence of toxin-producing strains was found among the Aeromonas isolates. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. The present review was constructed with a view to highlight the zoonotic importance of Aeromonas pathogen in fish and chicken meat. PMID:27051177

  18. The Main Aeromonas Pathogenic Factors

    PubMed Central

    Tomás, J. M.

    2012-01-01

    The members of the Aeromonas genus are ubiquitous, water-borne bacteria. They have been isolated from marine waters, rivers, lakes, swamps, sediments, chlorine water, water distribution systems, drinking water and residual waters; different types of food, such as meat, fish, seafood, vegetables, and processed foods. Aeromonas strains are predominantly pathogenic to poikilothermic animals, and the mesophilic strains are emerging as important pathogens in humans, causing a variety of extraintestinal and systemic infections as well as gastrointestinal infections. The most commonly described disease caused by Aeromonas is the gastroenteritis; however, no adequate animal model is available to reproduce this illness caused by Aeromonas. The main pathogenic factors associated with Aeromonas are: surface polysaccharides (capsule, lipopolysaccharide, and glucan), S-layers, iron-binding systems, exotoxins and extracellular enzymes, secretion systems, fimbriae and other nonfilamentous adhesins, motility and flagella. PMID:23724321

  19. Diversity, persistence, and virulence of Aeromonas strains isolated from drinking water distribution systems in Sweden.

    PubMed Central

    Kühn, I; Allestam, G; Huys, G; Janssen, P; Kersters, K; Krovacek, K; Stenström, T A

    1997-01-01

    The Aeromonas populations in 13 Swedish drinking water distribution systems, representing different treatments, were investigated. From each system, water samples were collected four times during the period from May to September 1994 from raw water and water after treatment and at two to five sites within the distribution system. In total, 220 water samples were collected. From samples containing presumptive Aeromonas, up to 32 colonies were analyzed by the PhenePlate Aeromonas (PhP-AE) system, which is a highly discriminating biochemical fingerprinting method. Selected isolates from different phenotypes (PhP types) were further identified by the API 20 NE system and by gas-liquid chromatography analysis of fatty acid methyl esters (FAMEs). Selected isolates were also assayed for their potential to produce hemolysin and cytotoxin and for their ability to adhere to human intestinal cells. In total, 117 water samples (53%) contained presumptive Aeromonas which numbered up to 10(6) CFU/100 ml in raw water and up to 750 CFU/100 ml in tap water. Among the 2,117 isolates that were subjected to typing by the PhP-AE system, more than 300 distinct PhP types were found, of which the majority occurred only sporadically. Raw (surface) water samples usually contained many different PhP types, showing high diversity indices (Di) (median Di = 0.95). The Aeromonas populations in samples collected from within the distribution systems were less diverse (median Di = 0.58) and were often dominated by one major PhP type that was found on several sampling occasions. Seventeen such major PhP types could be found and were represented in 1,037 isolates (49%). Identification by API 20 NE and FAME analysis revealed that most of the major PhP types were Aeromonas hydrophila or belonged to unidentified Aeromonas species. Hemolysin and cytotoxin production was observed in most major PhP types (representing 87 and 54% of the assayed isolates, respectively), and adherence was found in 89% of the

  20. Aldohexuronic Acid Catabolism by a Soil Aeromonas

    PubMed Central

    Farmer, J. J.; Eagon, R. G.

    1969-01-01

    Bacteria which utilize mannuronic acid as an energy source were isolated from nature. One of the organisms, identified as a member of the genus Aeromonas, used glucuronate, galacturonate, and mannuronate as the sole source of carbon and energy. Glucuronate- and galacturonate-grown resting cells oxidized both glucuronate and galacturonate rapidly, but mannuronate slowly. Mannuronate-grown cells oxidized all three rapidly, with the rate of mannuronate utilization somewhat lower. Cell-free extracts from glucuronate-, galacturonate-, and mannuronate-grown Aeromonas C11-2B contained glucuronate and galacturonate isomerases, fructuronate, tagaturonate, and mannuronate reductases, and mannonate and altronate dehydratases, with the exception of glucuronate-grown cells which lacked altronate dehydratase. Thus, the pathway for glucuronate and galacturonate catabolism for Aeromonas was identical to Escherichia coli. Glucuronate and galacturonate were isomerized to d-fructuronate and d-tagaturonate which were then reduced by reduced nicotinamide adenine dinucleotide to d-mannonate and d-altronate, respectively. The hexonic acids were dehydrated to 2-keto-3-deoxy gluconate which was phosphorylated by adenosine triphosphate to 2-keto-3-deoxy-6-phospho gluconate. The latter was then cleaved to pyruvate and glyceraldehyde-3-phosphate. Mannuronate was reduced directly to d-mannonate by a reduced nicotinamide adenine dinucleotide phosphate-linked oxidoreductase. d-Mannonate was then further broken down as in the glucuronate pathway. The mannuronate reducing enzyme, for which the name d-mannonate:nicotinamide adenine dinucleotide (phosphate) oxidoreductase (d-mannuronate-forming) was proposed, was shown to be distinct from altronate and mannoate oxidoreductases. This is the first report of a bacterial oxidoreductase which reduces an aldohexuronic acid to a hexonic acid. The enzyme should prove to be a useful analytical tool for determining mannuronate in the presence of other uronic

  1. [Aeromonas spp. and Plesiomonas shigelloides in bivalves, mud, and water from the Gulf of Nicoya, Costa Rica].

    PubMed

    Rodríguez, E; Antillón, F

    1989-06-01

    Bivalves, mud, and surface water were collected at three different sites of the Gulf of Nicoya, Costa Rica, in search of Aeromonas spp. and Plesiomonas shigelloides. For their isolation, these bacteria were enriched in alkaline peptone water and streaked on MacConkey agar and on brilliant green bile inositol agar. This was followed by the biochemical tests necessary for their identification. Thirty-five strains of A. hydrophila, 58 of A. caviae, 43 of A. sobria, and 7 of P. shigelloides were isolated. None of these predominated nor was there any indication of a seasonal distribution along the 15 month's duration of the study. Seven strains of A. hydrophila and two of A. sobria showed the biochemical characteristics associated with toxin production (positive Voges-Proskauer and lysine decarboxylase tests). These species are widely distributed in the gulf and there is risk of contracting an infection while bathing or when eating raw bivalves from this area.

  2. Multi-Drug Resistance Mediated by Class 1 Integrons in Aeromonas Isolated from Farmed Freshwater Animals

    PubMed Central

    Deng, Yuting; Wu, Yali; Jiang, Lan; Tan, Aiping; Zhang, Ruiquan; Luo, Li

    2016-01-01

    Aeromonas is regarded as an important pathogen of freshwater animals but little is known about the genetics of its antimicrobial resistance in Chinese aquaculture. The aim of this study was to investigate the presence of integrons and characterize multidrug resistant Aeromonas spp. isolated from diseased farmed freshwater animals. These animal samples included fish, ornamental fish, shrimp, turtles, and amphibians which were collected from 64 farms in Guangdong province of South China. One hundred and twelve Aeromonas spp. isolates were examined for antimicrobial resistance phenotypes and the presence of class 1 integron sequences. Twenty-two (19.6%) of these isolates carried a class 1 integron comprising six different gene insertion cassettes including drfA12-orfF-aadA2, drfA12-orfF, aac(6′)-II-blaOXA-21-cat3, catB3, arr-3, and dfrA17. Among these, drfA12-orfF-aadA2 was the dominant gene cassette array (63.6%, 14/22) and this is the first report of aac(6′)-II-blaOXA-21-cat3 in an Aeromonas hydrophila isolate from a Chinese giant salamander (Andrias davidianus). All the integron-positive strains were resistant to more than five agents and 22 contained other resistance genes including blaCTX-M-3, blaTEM-1, aac(6′)-Ib-cr, and tetA. All integron-positive isolates also contained mutations in the quinolone resistance determining regions (QRDR). Our investigation demonstrates that freshwater animals can serve as a reservoir for pathogenic Aeromonas strains containing multiple drug-resistance integrons. This data suggests that surveillance for antimicrobial resistance of animal origin and a prudent and responsible use of antimicrobials in aquaculture is necessary in these farms. PMID:27379065

  3. Quinolone resistant Aeromonas spp. as carriers and potential tracers of acquired antibiotic resistance in hospital and municipal wastewater.

    PubMed

    Varela, Ana Rita; Nunes, Olga C; Manaia, Célia M

    2016-01-15

    Members of the genus Aeromonas are recognized carriers of antibiotic resistance in aquatic environments. However, their importance on the spread of resistance from hospital effluents to the environment is poorly understood. Quinolone resistant Aeromonas spp. (n = 112) isolated from hospital effluent (HE) and from raw (RWW) and treated wastewater (TWW) of the receiving urban wastewater treatment plant (UWTP) were characterized. Species identification and genetic intraspecies diversity were assessed based on the 16S rRNA, cpn60 and gyrB genes sequence analysis. The antibiotic resistance phenotypes and genotypes (qnrA, qnrB, qnrC, qnrD, qnrS, qnrVC; qepA; oqxAB; aac(6′)-Ib-cr; blaOXA; incU) were analyzed in function of the origin and taxonomic group. Most isolates belonged to the species Aeromonas caviae and Aeromonas hydrophila (50% and 41%, respectively). The quinolone and the beta-lactamase resistance genes aac(6′)-Ib-cr and blaOXA, including gene blaOXA-101, identified for the first time in Aeromonas spp., were detected in 58% and 56% of the isolates, respectively, with identical prevalence in HE and UWTP wastewater. In contrast, the gene qnrS2 was observed mainly in isolates from the UWTP (51%) and rarely in HE isolates (3%), suggesting that its origin is not the clinical setting. Bacterial groups and genes that allow the identification of major routes of antibiotic resistance dissemination are valuable tools to control this problem. In this study, it was concluded that members of the genus Aeromonas harboring the genes aac(6′)-Ib-cr and blaOXA are relevant tracers of antibiotic resistance dissemination in wastewater habitats, while those yielding the gene qnrS2 allow the traceability from non-clinical sources.

  4. The Animal Model Determines the Results of Aeromonas Virulence Factors

    PubMed Central

    Romero, Alejandro; Saraceni, Paolo R.; Merino, Susana; Figueras, Antonio; Tomás, Juan M.; Novoa, Beatriz

    2016-01-01

    The selection of an experimental animal model is of great importance in the study of bacterial virulence factors. Here, a bath infection of zebrafish larvae is proposed as an alternative model to study the virulence factors of Aeromonas hydrophila. Intraperitoneal infections in mice and trout were compared with bath infections in zebrafish larvae using specific mutants. The great advantage of this model is that bath immersion mimics the natural route of infection, and injury to the tail also provides a natural portal of entry for the bacteria. The implication of T3SS in the virulence of A. hydrophila was analyzed using the AH-1::aopB mutant. This mutant was less virulent than the wild-type strain when inoculated into zebrafish larvae, as described in other vertebrates. However, the zebrafish model exhibited slight differences in mortality kinetics only observed using invertebrate models. Infections using the mutant AH-1ΔvapA lacking the gene coding for the surface S-layer suggested that this protein was not totally necessary to the bacteria once it was inside the host, but it contributed to the inflammatory response. Only when healthy zebrafish larvae were infected did the mutant produce less mortality than the wild-type. Variations between models were evidenced using the AH-1ΔrmlB, which lacks the O-antigen lipopolysaccharide (LPS), and the AH-1ΔwahD, which lacks the O-antigen LPS and part of the LPS outer-core. Both mutants showed decreased mortality in all of the animal models, but the differences between them were only observed in injured zebrafish larvae, suggesting that residues from the LPS outer core must be important for virulence. The greatest differences were observed using the AH-1ΔFlaB-J (lacking polar flagella and unable to swim) and the AH-1::motX (non-motile but producing flagella). They were as pathogenic as the wild-type strain when injected into mice and trout, but no mortalities were registered in zebrafish larvae. This study demonstrates

  5. Distribution of Virulence Factors and Molecular Fingerprinting of Aeromonas Species Isolates from Water and Clinical Samples: Suggestive Evidence of Water-to-Human Transmission ▿ †

    PubMed Central

    Khajanchi, Bijay K.; Fadl, Amin A.; Borchardt, Mark A.; Berg, Richard L.; Horneman, Amy J.; Stemper, Mary E.; Joseph, Sam W.; Moyer, Nelson P.; Sha, Jian; Chopra, Ashok K.

    2010-01-01

    A total of 227 isolates of Aeromonas obtained from different geographical locations in the United States and different parts of the world, including 28 reference strains, were analyzed to determine the presence of various virulence factors. These isolates were also fingerprinted using biochemical identification and pulse-field gel electrophoresis (PFGE). Of these 227 isolates, 199 that were collected from water and clinical samples belonged to three major groups or complexes, namely, the A. hydrophila group, the A. caviae-A. media group, and the A. veronii-A. sobria group, based on biochemical profiles, and they had various pulsotypes. When virulence factor activities were examined, Aeromonas isolates obtained from clinical sources had higher cytotoxic activities than isolates obtained from water sources for all three Aeromonas species groups. Likewise, the production of quorum-sensing signaling molecules, such as N-acyl homoserine lactone, was greater in clinical isolates than in isolates from water for the A. caviae-A. media and A. hydrophila groups. Based on colony blot DNA hybridization, the heat-labile cytotonic enterotoxin gene and the DNA adenosine methyltransferase gene were more prevalent in clinical isolates than in water isolates for all three Aeromonas groups. Using colony blot DNA hybridization and PFGE, we obtained three sets of water and clinical isolates that had the same virulence signature and had indistinguishable PFGE patterns. In addition, all of these isolates belonged to the A. caviae-A. media group. The findings of the present study provide the first suggestive evidence of successful colonization and infection by particular strains of certain Aeromonas species after transmission from water to humans. PMID:20154106

  6. CHARACTERIZATION OF PRECURSOR FOR 16S rRNA FOR AEROMONAS HYDROPHILA

    EPA Science Inventory

    Current strategies for monitoring drinking water quality involve culture-based methods to detect the presence of microbial indicators. However, these methods are insensitive when the organisms have undergone physiological changes such as injury and starvation that can occur in h...

  7. Evasion of mucosal defenses during Aeromonas hydrophila infection of channel catfish (Ictalurus punctatus) skin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The mucosal surfaces of fish serve as the first-line of defense against the myriad of aquatic pathogens present in the aquatic environment. The immune repertoire functioning at these interfaces is still poorly understood. The skin, in particular, must process signals from several fronts, sensing and...

  8. Purification and characterization of Aeromonas sobria pili, a possible colonization factor.

    PubMed Central

    Hokama, A; Iwanaga, M

    1991-01-01

    Pili of Aeromonas sobria Ae1 were purified and characterized. The molecular mass of the pilin was estimated to be about 23 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Ae1 pili were electrophoretically and immunologically distinguishable from the W pili of A. hydrophila Ae6, although the two pili were morphologically indistinguishable. The N-terminal amino acid sequences of the two pilins were identical in the first 10 residues. Strain Ae1 and its purified pili adhered to human and rabbit intestines and agglutinated human and rabbit erythrocytes. Hemagglutination was inhibited by D-galactose and D-mannose, but not by L-fucose. Organisms pretreated with the Fab fraction of the antipilus antibody failed to adhere to the intestines. Organisms did not adhere to intestines pretreated with the purified pili. These findings suggest that the pili are a colonization factor of A. sobria Ae1. Images PMID:1680105

  9. Aeromonas dhakensis pneumonia and sepsis in a neonate Risso's dolphin Grampus griseus from the Mediterranean Sea.

    PubMed

    Pérez, Lola; Abarca, M Lourdes; Latif-Eugenín, Fadua; Beaz-Hidalgo, Roxana; Figueras, M José; Domingo, Mariano

    2015-09-17

    A neonate Risso's dolphin Grampus griseus was found stranded alive on a beach in Catalonia, Spain. Rehabilitation attempts were unsuccessful and it died 2 d later, showing pneumonia and sepsis. A pure bacterial culture was obtained from all tissues and blood and identified as Aeromonas hydrophila using the API 20NE. However, sequencing the rpoD gene showed that the strain in fact belongs to A. dhakensis, making this the first report of fatal haemorrhagic-necrotizing pneumonia and sepsis due to this species in a marine mammal. The A. dhakensis strain GMV-704 produced β-haemolysis, possessed several virulence genes and showed sensitivity to several antimicrobials. This study provides a new potential host for A. dhakensis, and its potential virulence in dolphins and its presence in the marine environment may warrant considering this species a potential threat to marine mammals.

  10. Draft genome sequence of the Chilean isolate Aeromonas salmonicida strain CBA100.

    PubMed

    Valdes, Natalia; Espinoza, Carolina; Sanhueza, Loreto; Gonzalez, Alex; Corsini, Gino; Tello, Mario

    2015-03-01

    We report the draft genome sequence from Aeromonas salmonicida sp. strain CBA100, which was characterized as an antibiotic-resistant bacterium isolated from infected rainbow trout. The total size of the genome is 4,788,109 bp, with a G + C content of 60.55%. Comparison of its open reading frames shows that the closest homologue to one third of the genes of strain CBA100 are found in A. hydrophila. The strain contains several efflux pumps and putative genes that confer resistance to multiclass antibiotics, including macrolide, β-lactamics, florfenicol and quinolones. The antibiogram profile suggests that efflux pumps are the main mechanism of resistance to non-β-lactamic antibiotics. This is the first genome of a Chilean isolate of A. salmonicida, which should shed light on the design of strain-specific vaccines against this pathogen and reduce the use of antibiotics for preventive treatment in Chilean aquaculture.

  11. The Aeromonas salmonicida Lipopolysaccharide Core from Different Subspecies: The Unusual subsp. pectinolytica.

    PubMed

    Merino, Susana; Tomás, Juan M

    2016-01-01

    Initial hydridization tests using Aeromonas salmonicida typical and atypical strains showed the possibility of different lipopolysaccharide (LPS) outer cores among these strains. By chemical structural analysis, LPS-core SDS-PAGE gel migration, and functional and comparative genomics we demonstrated that typical A. salmonicida (subsp. salmonicida) strains and atypical subsp. masoucida and probably smithia strains showed the same LPS outer core. A. salmonicida subsp. achromogenes strains show a similar LPS outer core but lack one of the most external residues (a galactose linked α1-6 to heptose), not affecting the O-antigen LPS linkage. A. salmonicida subsp. pectinolytica strains show a rather changed LPS outer core, which is identical to the LPS outer core from the majority of the A. hydrophila strains studied by genomic analyses. The LPS inner core in all tested A. salmonicida strains, typical and atypical, is well-conserved. Furthermore, the LPS inner core seems to be conserved in all the Aeromonas (psychrophilic or mesophilic) strains studied by genomic analyses.

  12. sugE: A gene involved in tributyltin (TBT) resistance of Aeromonas molluscorum Av27.

    PubMed

    Cruz, Andreia; Micaelo, Nuno; Félix, Vitor; Song, Jun-Young; Kitamura, Shin-Ichi; Suzuki, Satoru; Mendo, Sónia

    2013-01-01

    The mechanism of bacterial resistance to tributyltin (TBT) is still unclear. The results herein presented contribute to clarify that mechanism in the TBT-resistant bacterium Aeromonas molluscorum Av27. We have identified and cloned a new gene that is involved in TBT resistance in this strain. The gene is highly homologous (84%) to the Aeromonas hydrophila-sugE gene belonging to the small multidrug resistance gene family (SMR), which includes genes involved in the transport of lipophilic drugs. In Av27, expression of the Av27-sugE was observed at the early logarithmic growth phase in the presence of a high TBT concentration (500 μM), thus suggesting the contribution of this gene for TBT resistance. E. coli cells transformed with Av27-sugE become resistant to ethidium bromide (EtBr), chloramphenicol (CP) and tetracycline (TE), besides TBT. According to the Moriguchi logP (miLogP) values, EtBr, CP and TE have similar properties and are substrates for the sugE-efflux system. Despite the different miLogP of TBT, E. coli cells transformed with Av27-sugE become resistant to this compound. So it seems that TBT is also a substrate for the SugE protein. The modelling studies performed also support this hypothesis. The data herein presented clearly indicate that sugE is involved in TBT resistance of this bacterium.

  13. Prevalence, virulence and antimicrobial resistance patterns of Aeromonas spp. isolated from children with diarrhea

    PubMed Central

    Soltan Dallal, Mohammad Mehdi; Mazaheri Nezhad Fard, Ramin; Kavan Talkhabi, Morteza; Aghaiyan, Leyla; Salehipour, Zohre

    2016-01-01

    Background Aeromonas spp. cause various intestinal and extraintestinal diseases. These bacteria are usually isolated from fecal samples, especially in children under five years old. The aim of this study was to assess the prevalence of Aeromonas spp. and their antimicrobial resistance profile in children with diarrhea referred to the Children Medical Center in Tehran, between 2013 and 2014. Methods A total number of 391 stool samples were collected from children with ages between 1 day and 14 years old, with diarrhea (acute or chronic), referred to the Children Hospital, Tehran, Iran, between 2013 and 2014. Samples were enriched in alkaline peptone water broth for 24 hours at 37 °C and then cultured. Suspicious colonies were analyzed through biochemical tests. Furthermore, antimicrobial susceptibility tests were carried out for the isolates. Isolates were further studied for act, ast, alt, aerA and hlyA virulence genes using polymerase chain reaction. Results In total, 12 isolates (3.1%) were identified as Aeromonas spp.; all were confirmed using the API-20E test. Of these isolates, five A. caviae (42%), four A. veronii (33%) and three A. hydrophila (25%) were identified in cases with gastroenteritis. Second to ampicillin (which was included in the growth medium used), the highest rate of antimicrobial resistance was seen against nalidixic acid and trimethoprim-sulfamethoxazole (5 isolates each, 41.6%) and the lowest rate of antimicrobial resistance was seen against gentamicin, amikacin and cefepime (none of the isolates). Results included 76.4% act, 64.7% ast, 71.5% alt, 83.3% aerA and 11.7% hlyA genes. Conclusion Aeromonas spp. are important due to their role in diarrhea in children; therefore, isolation and identification of these fecal pathogens should seriously be considered in medical laboratories. Since virulence genes play a significant role in gastroenteritis symptoms caused by these bacteria, Aeromonas species that include virulence genes are potentially

  14. Symbiosis of Aeromonas veronii Biovar sobria and Hirudo medicinalis, the Medicinal Leech: a Novel Model for Digestive Tract Associations

    PubMed Central

    Graf, Joerg

    1999-01-01

    Hirudo medicinalis, the medicinal leech, is applied postoperatively in modern medicine. Infections by Aeromonas occur in up to 20% of patients unless a preemptive antibiotic treatment is administered. The associated infections demonstrate the need for a better understanding of the digestive tract flora of H. medicinalis. Early studies reported the presence of a single bacterial species in the digestive tract and suggested that these bacteria were endosymbionts contributing to the digestion of blood. In this study, we cultivated bacteria from the digestive tract and characterized them biochemically. The biochemical test results identified the isolates as Aeromonas veronii biovar sobria. This species identification was supported by sequence comparison of a variable region of the genes coding for 16S rRNA. In a colonization assay, a rifampin-resistant derivative of a symbiotic isolate was fed in a blood meal to H. medicinalis. The strain colonized the digestive tract rapidly and reached a concentration similar to that of the native bacterial flora. For the first 12 h, the in vivo doubling time was 1.2 h at 23°C. After 12 h, at a density of 5 × 107 CFU/ml, the increase in viable counts ceased, suggesting a dramatic reduction in the bacterial growth rate. Two human fecal isolates, identified as Aeromonas hydrophila and A. veronii biovar sobria, were also able to colonize the digestive tract. These data demonstrate that the main culturable bacterium in the crop of H. medicinalis is A. veronii biovar sobria and that the medicinal leech can be used as a model for digestive tract association of Aeromonas species. PMID:9864188

  15. Active Shiga-Like Toxin Produced by Some Aeromonas spp., Isolated in Mexico City.

    PubMed

    Palma-Martínez, Ingrid; Guerrero-Mandujano, Andrea; Ruiz-Ruiz, Manuel J; Hernández-Cortez, Cecilia; Molina-López, José; Bocanegra-García, Virgilio; Castro-Escarpulli, Graciela

    2016-01-01

    Shiga-like toxins (Stx) represent a group of bacterial toxins involved in human and animal diseases. Stx is produced by enterohemorrhagic Escherichia coli, Shigella dysenteriae type 1, Citrobacter freundii, and Aeromonas spp.; Stx is an important cause of bloody diarrhea and hemolytic uremic syndrome (HUS). The aim of this study was to identify the stx1/stx2 genes in clinical strains and outer membrane vesicles (OMVs) of Aeromonas spp., 66 strains were isolated from children who live in Mexico City, and Stx effects were evaluated in Vero cell cultures. The capacity to express active Stx1 and Stx2 toxins was determined in Vero cell cultures and the concentration of Stx was evaluated by 50% lethal dose (LD50) assays, observing inhibition of damaged cells by specific monoclonal antibodies. The results obtained in this study support the hypothesis that the stx gene is another putative virulence factor of Aeromonas, and since this gene can be transferred horizontally through OMVs this genus should be included as a possible causal agents of gastroenteritis and it should be reported as part of standard health surveillance procedures. Furthermore, these results indicate that the Aeromonas genus might be a potential causative agent of HUS.

  16. Active Shiga-Like Toxin Produced by Some Aeromonas spp., Isolated in Mexico City

    PubMed Central

    Palma-Martínez, Ingrid; Guerrero-Mandujano, Andrea; Ruiz-Ruiz, Manuel J.; Hernández-Cortez, Cecilia; Molina-López, José; Bocanegra-García, Virgilio; Castro-Escarpulli, Graciela

    2016-01-01

    Shiga-like toxins (Stx) represent a group of bacterial toxins involved in human and animal diseases. Stx is produced by enterohemorrhagic Escherichia coli, Shigella dysenteriae type 1, Citrobacter freundii, and Aeromonas spp.; Stx is an important cause of bloody diarrhea and hemolytic uremic syndrome (HUS). The aim of this study was to identify the stx1/stx2 genes in clinical strains and outer membrane vesicles (OMVs) of Aeromonas spp., 66 strains were isolated from children who live in Mexico City, and Stx effects were evaluated in Vero cell cultures. The capacity to express active Stx1 and Stx2 toxins was determined in Vero cell cultures and the concentration of Stx was evaluated by 50% lethal dose (LD50) assays, observing inhibition of damaged cells by specific monoclonal antibodies. The results obtained in this study support the hypothesis that the stx gene is another putative virulence factor of Aeromonas, and since this gene can be transferred horizontally through OMVs this genus should be included as a possible causal agents of gastroenteritis and it should be reported as part of standard health surveillance procedures. Furthermore, these results indicate that the Aeromonas genus might be a potential causative agent of HUS. PMID:27725813

  17. Aeromonas flagella and colonisation mechanisms.

    PubMed

    Lowry, Rebecca; Balboa, Sabela; Parker, Jennifer L; Shaw, Jonathan G

    2014-01-01

    Aeromonas species are inhabitants of aquatic environments and are able to cause disease in humans and fish among other animals. In aquaculture, they are responsible for the economically important diseases of furunculosis and motile Aeromonas septicaemia (MAS). Whereas gastroenteritis and wound infections are the major human diseases associated with the genus. As they inhabit and survive in diverse environments, aeromonads possess a wide range of colonisation factors. The motile species are able to swim in liquid environments through the action of a single polar flagellum, the flagellin subunits of which are glycosylated; although essential for function the biological role of glycan addition is yet to be determined. Approximately 60% of aeromonads possess a second lateral flagella system that is expressed in viscous environments for swarming over surfaces; both flagellar systems have been shown to be important in the initial colonisation of surfaces. Subsequently, other non-flagellar colonisation factors are employed; these can be both filamentous and non-filamentous. The aeromonads possess a number of fimbrial systems with the bundle-forming MSHA type IV pilus system, having a major role in human cell adherence. Furthermore, a series of outer-membrane proteins have also been implicated in the aeromonad adhesion process. A number of strains are also capable of cell invasion and that maybe linked with the more invasive diseases of bacteraemia or wound infections. These strains employ cell surface factors that allow the colonisation of these niches that protect them from the host's immune system such as S-layers, capsules or particular lipopolysaccharides.

  18. Characterization of integrons and tetracycline resistance determinants in Aeromonas spp. isolated from South African aquaculture systems.

    PubMed

    Jacobs, Liezl; Chenia, Hafizah Y

    2007-03-20

    An increasing incidence of multidrug resistance amongst Aeromonas spp. isolates, which are both fish pathogens and emerging opportunistic human pathogens, has been observed worldwide. This can be attributed to the horizontal transfer of mobile genetic elements, viz.: plasmids and class 1 integrons. The antimicrobial susceptibilities of 37 Aeromonas spp. isolates, from tilapia, trout and koi aquaculture systems, were determined by disc-diffusion testing. The plasmid content of each isolate was examined using the alkaline lysis protocol. Tet determinant type was determined by amplification using two degenerate primer sets and subsequent HaeIII restriction. The presence of integrons was determined by PCR amplification of three integrase genes, as well as gene cassettes, and the qacEDelta1-sulI region. Thirty-seven Aeromonas spp. isolates were differentiated into six species by aroA PCR-RFLP, i.e., A. veronii biovar sobria, A. hydrophila, A. encheleia, A. ichtiosoma, A. salmonicida, and A. media. High levels of resistance to tetracycline (78.3%), amoxicillin (89.2%), and augmentin (86.5%) were observed. Decreased susceptibility to erythromycin was observed for 67.6% of isolates. Although 45.9% of isolates displayed nalidixic acid resistance, majority of isolates were susceptible to the fluoroquinolones. The MAR index ranged from 0.12 to 0.59, with majority of isolates indicating high-risk contamination originating from humans or animals where antibiotics are often used. Plasmids were detected in 21 isolates, with 14 of the isolates displaying multiple plasmid profiles. Single and multiple class A family Tet determinants were observed in 27% and 48.7% of isolates, respectively, with Tet A being the most prevalent Tet determinant type. Class 1 integron and related structures were amplified and carried different combinations of the antibiotic resistance gene cassettes ant(3'')Ia, aac(6')Ia, dhfr1, oxa2a and/or pse1. Class 2 integrons were also amplified, but the

  19. Draft Genome Sequence of Aeromonas sp. Strain EERV15

    PubMed Central

    Ehsani, Elham; Barrantes, Israel; Vandermaesen, Johanna; Geffers, Robert; Jarek, Michael; Boon, Nico; Springael, Dirk; Pieper, Dietmar H.

    2016-01-01

    We report here the draft genome sequence of Aeromonas sp. strain EERV15 isolated from sand filter. The organism most closely related to Aeromonas sp. EERV15 is Aeromonas veronii B565, with an average 83% amino acid sequence similarity of putatively encoded protein open reading frames. PMID:27540061

  20. Two Catechol Siderophores, Acinetobactin and Amonabactin, Are Simultaneously Produced by Aeromonas salmonicida subsp. salmonicida Sharing Part of the Biosynthetic Pathway.

    PubMed

    Balado, Miguel; Souto, Alba; Vences, Ana; Careaga, Valeria P; Valderrama, Katherine; Segade, Yuri; Rodríguez, Jaime; Osorio, Carlos R; Jiménez, Carlos; Lemos, Manuel L

    2015-12-18

    The iron uptake mechanisms based on siderophore synthesis used by the fish pathogen Aeromonas salmonicida subsp. salmonicida are still not completely understood, and the precise structure of the siderophore(s) is unknown. The analysis of genome sequences revealed that this bacterium possesses two gene clusters putatively involved in the synthesis of siderophores. One cluster is a candidate to encode the synthesis of acinetobactin, the siderophore of the human pathogen Acinetobacter baumannii, while the second cluster shows high similarity to the genes encoding amonabactin synthesis in Aeromonas hydrophila. Using a combination of genomic analysis, mutagenesis, biological assays, chemical purification, and structural determination procedures, here we demonstrate that most A. salmonicida subsp. salmonicida strains produce simultaneously the two siderophores, acinetobactin and amonabactin. Interestingly, the synthesis of both siderophores relies on a single copy of the genes encoding the synthesis of the catechol moiety (2,3-dihydroxybenzoic acid) and on one encoding a phosphopantetheinyl transferase. These genes are present only in the amonabactin cluster, and a single mutation in any of them abolishes production of both siderophores. We could also demonstrate that some strains, isolated from fish raised in seawater, produce only acinetobactin since they present a deletion in the amonabactin biosynthesis gene amoG. Our study represents the first evidence of simultaneous production of acinetobactin and amonabactin by a bacterial pathogen and reveals the plasticity of bacterial genomes and biosynthetic pathways. The fact that the same siderophore is produced by unrelated pathogens highlights the importance of these systems and their interchangeability between different bacteria.

  1. Aeromonas salmonicida grown in vivo.

    PubMed Central

    Garduño, R A; Thornton, J C; Kay, W W

    1993-01-01

    The virulent fish pathogen Aeromonas salmonicida was rapidly killed in vivo when restricted inside a diffusion chamber implanted intraperitoneally in rainbow trout. After a period of regrowth, the survivors had acquired resistance to host-mediated bacteriolysis, phagocytosis, and oxidative killing, properties which were subsequently lost by growth in vitro. Resistance to bacteriolysis and phagocytosis was associated with a newly acquired capsular layer revealed by acidic polysaccharide staining and electron microscopy. This capsular layer shielded the underlying, regular surface array (S-layer) from immunogold labeling with a primary antibody to the S-layer protein. Resistance to oxidative killing was mediated by a mechanism not associated with the presence of the capsular layer. An attenuated vaccine strain of A. salmonicida grown in vivo failed to express the capsular layer. Consequently, the in vivo-grown cells of this attenuated strain remained as sensitive to bacteriolysis, and as avidly adherent to macrophages, as the in vitro-grown cells. The importance of these new virulence determinants and their relation to the known virulence factors of A. salmonicida are discussed. Images PMID:8359906

  2. Aeromonas phages encode tRNAs for their overused codons.

    PubMed

    Prabhakaran, Ramanandan; Chithambaram, Shivapriya; Xia, Xuhua

    2014-01-01

    The GC-rich bacterial species, Aeromonas salmonicida, is parasitised by both GC-rich phages (Aeromonas phages - phiAS7 and vB_AsaM-56) and GC-poor phages (Aeromonas phages - 25, 31, 44RR2.8t, 65, Aes508, phiAS4 and phiAS5). Both the GC-rich Aeromonas phage phiAS7 and Aeromonas phage vB_AsaM-56 have nearly identical codon usage bias as their host. While all the remaining seven GC-poor Aeromonas phages differ dramatically in codon usage from their GC-rich host. Here, we investigated whether tRNA encoded in the genome of Aeromonas phages facilitate the translation of phage proteins. We found that tRNAs encoded in the phage genome correspond to synonymous codons overused in the phage genes but not in the host genes.

  3. [Prevalence of Aeromonas spp. in surface water].

    PubMed

    Hernández, P; Rodríguez de García, R

    1997-03-01

    Some Aeromonas strains are well recognized enteropathogens according to microbiological, clinical, immunological and epidemiological evidence. The main source of infection seems to be untreated water, these microorganisms can be found in virtually all aquatic environments. Additionally, some Aeromonas, which include enterotoxigenic strains, are capable of rapid growth at 5 degrees C and even of producing toxins. Vegetable products irrigated with contaminated water may reach critical Aeromonas levels after being kept under refrigeration, this could represent a public health risk when they are consumed as uncooked salads. This study was pursued to evaluate such risk. Surface water samples were streaked on starch ampicillin and inositol-brilliant green-bile salts agar dishes. In addition, 100 ml of each sample were filtered through a 0.45 micron Millipore membrane filter. The filters were incubated on alkaline peptone water as enrichment media during 24 h at 35 degrees C. Enrichment broth was then streaked on the selective agars above mentioned. Isolates from both tests were identified using the API 20 E System. The prevalence of Aeromonas strains in the analyzed samples was 17.8%. A higher isolation rate was observed after the enrichment technique. Starch ampicillin agar showed a higher recuperation rate. A Veronii biotype sobria (formerly A. sobria) was isolated with higher frequency. Since this species has been associated with the greatest virulence, the use of contaminated water to irrigate vegetable products that are to be kept under refrigeration and consumed without ulterior cooking may represent a risk to the public health.

  4. Recombinant goose-type lysozyme in channel catfish: Lysozyme activity and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were: 1) to investigate whether recombinant channel catfish lysozyme g (CC-Lys-g) produced in E. coli expression system possesses any lysozyme activity; and 2) to evaluate whether channel catfish lysozyme g plasmid DNA could be used as an immunostimulant to protect chann...

  5. Recombinant goose-type lysozyme in channel catfish: lysozyme activity and efficacy as plasmid DNA immunostimulant against Aeromonas hydrophila infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were: 1) to investigate whether recombinant channel catfish lysozyme g (CC-Lys-g) produced in E. coli expression system possesses any lysozyme activity; and 2) to evaluate whether channel catfish lysozyme g plasmid DNA could be used as an immunostimulant to protect chann...

  6. An in vitro screening method to evaluate chemicals as potential chemotherapeutants to control Aeromonas hydrophila infection in channel catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using catfish gill cells G1B and four chemicals (hydrogen peroxide, sodium chloride, potassium permanganate, and D-mannose), the feasibility of using an in vitro screening method to identify potential effective chemotherapeutants was evaluated in this study. In vitro screening results revealed that,...

  7. Edwardsiella tarda and Aeromonas hydrophila isolated from diseased Southern flounder (Paralichthys lethostigma) are virulent to channel catfish and Nile tilapia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The aim of this study is to identify bacterial pathogens isolated from diseased Southern flounder and determine their virulence to channel catfish and Nile tilapia. Twenty five Gram-negative bacteria isolates were recovered from five tissues (skin lesions, brain, liver, intestine, and posterior kidn...

  8. Virulence factors in clinical and food isolates of Aeromonas species.

    PubMed

    Pin, C; Marín, M L; Selgas, M D; García, M L; Tormo, J; Casas, C

    1994-01-01

    Virulence factors were compared in 15 Aeromonas spp. isolated from faeces of patients with Aeromonas-associated gastroenteritis and in 81 strains isolated from food. Strains from food did not show differences in the distribution of virulence factors when compared with strains isolated from faeces. However, 88.8% of Aeromonas strains isolated from food were capable of producing possible virulence factors. Characterization of 28 autoagglutinating (AA+) Aeromonas spp. indicated that the human strains differed from the food strains in hemagglutinating and hemolytic capacities. These results suggest that autoagglutination associated with hemagglutinating and hemolytic capacities in food strains may be a helpful indicator of potential pathogenicity.

  9. High Prevalence of BlaCTX-M Group Genes in Aeromonas dhakensis Isolated from Aquaculture Fish Species in South Korea

    PubMed Central

    YI, Seung-Won; CHUNG, Tae-Ho; JOH, Seong-Joon; PARK, Chul; PARK, Byoung-Yong; SHIN, Gee-Wook

    2014-01-01

    The prevalence of resistant genes against β-lactams in 119 Aeromonas strains was determined. A large number (99.2%) of the present fish strains were resistant to one or more β- lactams including ceftiofur, amoxicillin-clavulanic acid, ampicillin, piperacillin and cefpodoxime. Among antibiotic resistance phenotypes, the simultaneous resistance to all β-lactams occurred in 25.2% (n=30) of all strains, which consisted of 18 strains of A. dhakensis, 8 strains of A. caviae, 2 strains of A. hydrophila and only one strain of A. veronii. For exploring genetic background of the antibiotic resistances, multiple PCR assays were subjected to detect β-lactamase-encoding genes, blaTEM, blaOXA-B and blaCTX-M. In the results, the blaTEM-1 gene was harbored in all strains, whereas only 3 strains harbored blaOXA gene. In the case of blaCTX-M gene, the gene was detected in 21.0% (25 out of 119) of all strains, which countered with 80% (20 out of 25) of A. dhakensis, 8% (2 out of 25) of A. caviae and 12% (3 out of 25) of A. hydrophila. In addition, most of the blaCTX-M positive strains showed simultaneous resistance to all β-lactams (18 out of 30 strains). In sequence analysis for blaCTX-M genes detected, they were CTX-M group 1-encoding genes including blaCTX-M-33 from 3 eel strains of A. dhakensis. Therefore, A. dhakensis obtained from cultured fish could represent a reservoir for spreading genes encoding CTX-M group 1 enzymes and hence should be carefully monitored, especially for its potential risk to public health. PMID:25649940

  10. High prevalence of blaCTX-M group genes in Aeromonas dhakensis isolated from aquaculture fish species in South Korea.

    PubMed

    Yi, Seung-Won; Chung, Tae-Ho; Joh, Seong-Joon; Park, Chul; Park, Byoung-Yong; Shin, Gee-Wook

    2014-12-01

    The prevalence of resistant genes against β-lactams in 119 Aeromonas strains was determined. A large number (99.2%) of the present fish strains were resistant to one or more β- lactams including ceftiofur, amoxicillin-clavulanic acid, ampicillin, piperacillin and cefpodoxime. Among antibiotic resistance phenotypes, the simultaneous resistance to all β-lactams occurred in 25.2% (n=30) of all strains, which consisted of 18 strains of A. dhakensis, 8 strains of A. caviae, 2 strains of A. hydrophila and only one strain of A. veronii. For exploring genetic background of the antibiotic resistances, multiple PCR assays were subjected to detect β-lactamase-encoding genes, bla(TEM), bla(OXA-B) and bla(CTX-M). In the results, the bla(TEM-1) gene was harbored in all strains, whereas only 3 strains harbored bla(OXA) gene. In the case of bla(CTX-M) gene, the gene was detected in 21.0% (25 out of 119) of all strains, which countered with 80% (20 out of 25) of A. dhakensis, 8% (2 out of 25) of A. caviae and 12% (3 out of 25) of A. hydrophila. In addition, most of the bla(CTX-M) positive strains showed simultaneous resistance to all β-lactams (18 out of 30 strains). In sequence analysis for bla(CTX-M) genes detected, they were CTX-M group 1-encoding genes including bla(CTX-M-33) from 3 eel strains of A. dhakensis. Therefore, A. dhakensis obtained from cultured fish could represent a reservoir for spreading genes encoding CTX-M group 1 enzymes and hence should be carefully monitored, especially for its potential risk to public health.

  11. An alternative bacteriological medium for the isolation of Aeromonas spp.

    USGS Publications Warehouse

    Jenkins, J.A.; Taylor, P.W.

    1995-01-01

    Two solid bacteriologic media were compared for cultivating Aeromonas spp. from piscine sources: the Rimler-Shotts (RS) medium and a starch-glutamate-ampicillin-penicillin-based medium (SGAP-10C) used for the recovery of Aeromonas spp. from water samples. The selective and differential capacities of the media were assessed March through October 1992 by recovery rate and phenotype of 99 isolates representing 15 genera of bacteria. Recovery frequency of Aeromonas spp. (n = 62) was similar at 97% on RS and 95% on SGAP-10C. The SGAP-10C medium proved to be more specific than RS toward Aeromonas species (P ≤ 0.005). Use of SGAP-10C at 24 C for 48 hr offers a better choice for the laboratory recovery of Aeromonas spp. from clinical fish specimens.

  12. Insight into the mobilome of Aeromonas strains

    PubMed Central

    Piotrowska, Marta; Popowska, Magdalena

    2015-01-01

    The mobilome is a pool of genes located within mobile genetic elements (MGE), such as plasmids, IS elements, transposons, genomic/pathogenicity islands, and integron-associated gene cassettes. These genes are often referred to as “flexible” and may encode virulence factors, toxic compounds as well as resistance to antibiotics. The phenomenon of MGE transfer between bacteria, known as horizontal gene transfer (HGT), is well documented. The genes present on MGE are subject to continuous processes of evolution and environmental changes, largely induced or significantly accelerated by man. For bacteria, the only chance of survival in an environment contaminated with toxic chemicals, heavy metals and antibiotics is the acquisition of genes providing the ability to survive in such conditions. The process of acquiring and spreading antibiotic resistance genes (ARG) is of particular significance, as it is important for the health of humans and animals. Therefore, it is important to thoroughly study the mobilome of Aeromonas spp. that is widely distributed in various environments, causing many diseases in fishes and humans. This review discusses the recently published information on MGE prevalent in Aeromonas spp. with special emphasis on plasmids belonging to different incompatibility groups, i.e., IncA/C, IncU, IncQ, IncF, IncI, and ColE-type. The vast majority of plasmids carry a number of different transposons (Tn3, Tn21, Tn1213, Tn1721, Tn4401), the 1st, 2nd, or 3rd class of integrons, IS elements (e.g., IS26, ISPa12, ISPa13, ISKpn8, ISKpn6) and encode determinants such as antibiotic and mercury resistance genes, as well as virulence factors. Although the actual role of Aeromonas spp. as a human pathogen remains controversial, species of this genus may pose a serious risk to human health. This is due to the considerable potential of their mobilome, particularly in terms of antibiotic resistance and the possibility of the horizontal transfer of resistance genes

  13. Occurrence and antibiotic resistance of mesophilic Aeromonas in three riverine freshwaters of Marrakech, Morocco.

    PubMed

    Imziln, B

    2001-12-01

    In order to evaluate the impact of pollution and sewage on the occurrence and antibiotic resistance of mesophilic aeromonads in riverine freshwaters of Marrakech, samples were collected from three rivers (Oukaimeden, Ourika, and Tensift) upstream and downstream from the principal bordering villages. During a 2-year study, indicators of pollution increased dramatically in the downstream waters. Bacterial indicators (faecal coliforms and faecal streptococci) correlated with mesophilic aeromonads only in heavily polluted waters. In low and moderately polluted sources, densities of mesophilic aeromonads were independent of water quality indicators and did not correlate statistically with faecal indicators. Average counts of Aeromonas in low and heavily polluted waters were 2.5 x 10(3) and 2.1 x 10(6) colony forming units per 100 ml, respectively. The biochemical identification of 841 isolates indicated a predominance of A. caviae in heavily and moderately polluted water and sediment. A. hydrophila was dominant only in low polluted waters and when the temperature was below 12 degrees C. High densities of A. sobria were found in low, moderately polluted, or cleaned waters and when the water temperature was above 18 degrees C. All selected isolates (total = 841) were tested for antibiotic susceptibility against 21 antibiotics. Antibiotic resistance frequencies recorded were: ampicillin and amoxicillin, 100%; novobiocin, 96%; cefalotin, 81%; colistin, 72%; sulfamethoxazole, 40%; cefamandole, 37%; polymyxin B, 23%; trimethoprim, 17%; erythromycin, 15%; streptomycin, 8%; amoxicillin-clavulanate, 5%. Resistance to cefotaxime, kanamycin, gentamycin, chloramphenicol, tetracycline, oxytetracycline, nalidixic acid, rifampicin, or trimethoprim-sulfameth-oxazole was found to be <5%. Antibiotic resistance rates did vary according to the source of a strain"s isolation, and high numbers of antibiotic resistant strains were recorded in polluted samples. Since no correlation between

  14. Vibrio cholerae and Aeromonas: do they share a mutual host?

    PubMed

    Senderovich, Yigal; Gershtein, Yana; Halewa, Etti; Halpern, Malka

    2008-03-01

    Species of the genus Aeromonas are native inhabitants of aquatic environments and have recently been considered as an emergent human pathogen. It is estimated that aeromonads cause up to 13% of reported gastroenteritis cases in the United States. Although the autochthonous existence of Aeromonas in the aquatic environment has been established, its natural reservoir is as yet unknown. Chironomids are closely related to mosquitoes except they do not bite and they are the most widely distributed insects in freshwater. They infest drinking water systems in Israel and all over the world. Vibrio cholerae inhabit chironomids and are able to degrade their egg masses. The degradation of the egg masses is followed by failure of the eggs to hatch. In the current study, egg masses from a waste stabilization pond and a river in northern Israel were collected and cultured during a five-month period. Bacterial colonies were randomly chosen and checked for their egg mass degradation abilities. In addition to V. cholerae, most of the other isolates that had the ability to degrade the egg masses were identified as Aeromonas species, thus, demonstrating that Aeromonas species are natural inhabitants of chironomid egg masses. The following virulence-associated genes were detected in Aeromonas species that were isolated from chironomid egg masses: alt (78%); ahpB (76%); act/aerA/hlyA (65%); fla (59%); pla/lipH3/apl-1/lip (43%); and ast (2%). These findings indicate that the Aeromonas species inhabiting chironomid egg masses pose a potential health risk. Understanding the natural reservoir of Aeromonas will help to develop methods to monitor and control the bacteria in fresh and drinking water reservoirs and to better understand the relationships between chironomids, V. cholerae and Aeromonas populations.

  15. Aeromonas Caviae Strain Induces Th1 Cytokine Response in Mouse Intestinal Tract

    EPA Science Inventory

    Aeromonas caviae has been associated with human gastrointestinal disease. Strains of this species typically lack virulence factors (VFs) such as enterotoxins and hemolysins that are produced by other human pathogens of the Aeromonas genus. Microarray profiling of murine small i...

  16. Biological characterization of Aeromonas spp. isolated from the environment.

    PubMed Central

    Rahim, Z.; Khan, S. I.; Chopra, A. K.

    2004-01-01

    Cytotoxic enterotoxin (Act) is a key virulence factor in the pathogenesis of infections caused by Aeromonas spp. The cytotoxic enterotoxin gene (act) was detected in 32 out of 69 environmental isolates of Aeromonas spp. by hybridization with the act gene probe. To evaluate the pathogenic potential of the act gene probe-positive isolates, 32 act gene probe-positive and 31 randomly selected act gene probe-negative isolates were tested for enterotoxicity in a suckling mice assay (SMA), for haemolytic activity on sheep blood agar plates, for the presence of CAMP-like factors, and for cytotoxicity in a Vero cell line. The act gene probe-positive isolates significantly differed from the toxin gene probe-negative ones with respect to enterotoxicity in the SMA (P=0.009) and haemolytic activity (P=0.005). The CAMP-haemolysin phenotype was significantly associated with the rabbit ileal loop assay (P= 0.08), Vero cell assay (P= 0.064), and haemolysin production under the microaerophilic conditions (P= 0.056) of the act gene probe-positive isolates of Aeromonas spp. These data indicated the role of Act in the pathogenesis of Aeromonas infections and that the enterotoxic potential of Aeromonas spp. could be assessed by simply performing a CAMP-haemolysin assay. PMID:15310164

  17. Clustering subspecies of Aeromonas salmonicida using IS630 typing

    PubMed Central

    2013-01-01

    Background The insertion element IS630 found in Aeromonas salmonicida belongs to the IS630-Tc1-mariner superfamily of transposons. It is present in multiple copies and represents approximately half of the IS present in the genome of A. salmonicida subsp. salmonicida A449. Results By using High Copy Number IS630 Restriction Fragment Length Polymorphism (HCN-IS630-RFLP), strains of various subspecies of Aeromonas salmonicida showed conserved or clustering patterns, thus allowing their differentiation from each other. Fingerprints of A. salmonicida subsp. salmonicida showed the highest homogeneity while ‘atypical’ A. salmonicida strains were more heterogeneous. IS630 typing also differentiated A. salmonicida from other Aeromonas species. The copy number of IS630 in Aeromonas salmonicida ranges from 8 to 35 and is much lower in other Aeromonas species. Conclusions HCN-IS630-RFLP is a powerful tool for subtyping of A. salmonicida. The high stability of IS630 insertions in A. salmonicida subsp. salmonicida indicates that it might have played a role in pathoadaptation of A. salmonicida which has reached an optimal configuration in the highly virulent and specific fish pathogen A. salmonicida subsp. salmonicida. PMID:23406017

  18. High-yield production of a chitinase from Aeromonas veronii B565 as a potential feed supplement for warm-water aquaculture.

    PubMed

    Zhang, Yuting; Zhou, Zhigang; Liu, Yuchun; Cao, Yanan; He, Suxu; Huo, Fengmin; Qin, Chubin; Yao, Bin; Ringø, Einar

    2014-02-01

    Chitin, present in crustacean shells, insects, and fungi, is the second most plentiful natural organic fiber after wood. To effectively use chitin in a cost-saving and environmentally friendly way in aquaculture, crustacean shells (e.g., shrimp-shell meal) are supplemented into aquafeed after degradation by chemical methods. Herein, we describe a chitinase from Aeromonas veronii B565, designated ChiB565, which potently degrades shrimp-shell chitin and resists proteolysis. We isolated recombinant ChiB565 of the expected molecular mass in large yield from Pichia pastoris. ChiB565 is optimally active at pH 5.0 and 50 °C and stable between pH 4.5 and 9.0 at 50 °C and below. Compared with the commercial chitinase C-6137, which cannot degrade shrimp-shell chitin, ChiB565 hydrolyzes shrimp-shell chitin in addition to colloidal chitin, powdered chitin, and β-1,3-1,4-glucan. The optimal enzyme concentration and reaction time for in vitro degradation of 0.1 g of powdered shrimp shell are 30 U of ChiB565 and 3 h, respectively. A synergistic protein-release effect occurred when ChiB565 and trypsin were incubated in vitro with shrimp shells. Tilapia were fed an experimental diet containing 5% (w/w) shrimp bran and 16.2 U/kg ChiB565, which significantly improved growth and feed conversion compared with a control diet lacking ChiB565. Dietary ChiB565 enhanced nitrogen digestibility and downregulated intestinal IL-1β expression. The immunologically relevant protective effects of dietary ChiB565 were also observed for 2 to 3 days following exposure to pathogenic Aeromonas hydrophila.

  19. Oral Challenge with Aeromonas in Protein-Malnourished Mice

    DTIC Science & Technology

    1994-01-01

    precipitaion after boiling. SRBC. sheep diarrhoea (RITARD) model (25) have red blood cells (hmolysis): Yt. adrenal cell culture; CHO. Chinese hamster...did not seem to affect the susceptibility of mice to deficient substitute for colostrum was not available. Aeromonas spp. infection, but there may be

  20. SENSITIVITY OF DIFFERENT AEROMONAS SPECIES TO COPPER AND SILVER

    EPA Science Inventory

    Aeromonas bacteria are common flora in surface and ground waters and are considered to be human pathogens. They can also be found in municipally treated drinking water, likely as a component of biofilms, as found in distribution system pipes and point of use water filters. It ...

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

    PubMed

    Davis, W A; Chretien, J H; Garagusi, V F; Goldstein, M A

    1978-04-01

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

  2. Viscoelastic properties of an exopolysaccharide: Aeromonas gum, produced by Aeromonas nichidenii 5797.

    PubMed

    Xu, Xiaojuan; Chen, Pan; Zhang, Lina

    2007-01-01

    The viscoelastic properties of Aeromonas (A) gum in water were investigated by using the Rheometric Scientific ARES controlled strain rheometer. An intrinsic viscosity of 8336 ml/g was obtained according to the Fuoss-Straus equation. The effect of salt concentration on intrinsic viscosity revealed that the A gum exists as semiflexible chain. Typical shear-thinning (pseudoplastic) behavior was observed at concentrations higher than 0.52%. The zero shear viscosity (eta(0)) increased with increasing polysaccharide concentration (c) showing a gradient of approximately 1.0, 2.9 and 4.8 in different concentration domains. The critical concentrations c* and c**, at which the transitions from a dilute solution of independently moving chains to semidilute and then concentrated domains occurred, were determined roughly to be 1.2% and 3.5%. The results from dynamic experiments revealed that the A gum solution shows characteristics of polymer solutions without any evidence of gel-like character. All the results from steady and dynamic tests suggest that the A gum is a non-gelling polysaccharide. The temperature dependence of apparent viscosity was described by Arrhenius equation and the flow activation energy was estimated to be 45.2 kJ/mol, which is independent on polymer concentration.

  3. Evolutionary Roots and Diversification of the Genus Aeromonas

    PubMed Central

    Sanglas, Ariadna; Albarral, Vicenta; Farfán, Maribel; Lorén, J. G.; Fusté, M. C.

    2017-01-01

    Despite the importance of diversification rates in the study of prokaryote evolution, they have not been quantitatively assessed for the majority of microorganism taxa. The investigation of evolutionary patterns in prokaryotes constitutes a challenge due to a very scarce fossil record, limited morphological differentiation and frequently complex taxonomic relationships, which make even species recognition difficult. Although the speciation models and speciation rates in eukaryotes have traditionally been established by analyzing the fossil record data, this is frequently incomplete, and not always available. More recently, several methods based on molecular sequence data have been developed to estimate speciation and extinction rates from phylogenies reconstructed from contemporary taxa. In this work, we determined the divergence time and temporal diversification of the genus Aeromonas by applying these methods widely used with eukaryotic taxa. Our analysis involved 150 Aeromonas strains using the concatenated sequences of two housekeeping genes (approximately 2,000 bp). Dating and diversification model analyses were performed using two different approaches: obtaining the consensus sequence from the concatenated sequences corresponding to all the strains belonging to the same species, or generating the species tree from multiple alignments of each gene. We used BEAST to perform a Bayesian analysis to estimate both the phylogeny and the divergence times. A global molecular clock cannot be assumed for any gene. From the chronograms obtained, we carried out a diversification analysis using several approaches. The results suggest that the genus Aeromonas began to diverge approximately 250 millions of years (Ma) ago. All methods used to determine Aeromonas diversification gave similar results, suggesting that the speciation process in this bacterial genus followed a rate-constant (Yule) diversification model, although there is a small probability that a slight

  4. Dietary β-glucans differentially modulate immune and stress-related gene expression in lymphoid organs from healthy and Aeromonas hydrophila-infected rainbow trout (Oncorhynchus mykiss).

    PubMed

    Douxfils, Jessica; Fierro-Castro, Camino; Mandiki, S N M; Emile, Wakson; Tort, Lluis; Kestemont, Patrick

    2017-04-01

    Although β-glucans stimulating effects have already been demonstrated on the immune system of numerous animal species, available data remain relatively variable and more research should be done regarding the complexity of underlying mechanisms. In this context, the present study aimed to evaluate the stress and immune-related effects of dietary β-glucans (i.e. Macrogard(®)) by considering a number of influencing factors such as the dose (0, 0.1, 0.2 and 0.5% in food), feeding duration (15 versus 30 days), tissue (blood, kidney, spleen, gills) and infection status (healthy or infected). Blood parameters (lysozyme, ACH50 activities, leucocyte populations) and mRNA expression level of several immune- and stress-related genes (TFN-α1, IL-1β, IL10, COX-2, TGF-β, MC2R, HSP70) were measured. Our results suggest that spleen may be a highly responsive organ to dietary β-glucans both in healthy or infected fish, and that this organ may therefore significantly contribute to the immune reinforcement induced by such immunostimulatory diet. Our study further reveals that overdoses of β-glucans and/or prolonged medication can lead to a non-reactive physiological status and, consequently, to a poor immune response. All in all, the current data emphasizes the need for further extensive research in the field of dietary β-glucans as a preventive method for farmed fish protection.

  5. Subunit vaccine candidates against Aeromonas salmonicida in rainbow trout Oncorhynchus mykiss

    PubMed Central

    Jørgensen, Louise von Gersdorff; Skov, Jakob; Chettri, Jiwan Kumar; Holm Mattsson, Andreas; Dalsgaard, Inger; Kania, Per Walter; Buchmann, Kurt

    2017-01-01

    Aeromonas salmonicida subsp. salmonicida is the etiological agent of furunculosis and a major fish health problem in salmonid aquaculture worldwide. Injection vaccination with commercial mineral oil-adjuvanted bacterin vaccines has been partly successful in preventing the disease but in Danish rainbow trout (Oncorhynchus mykiss, Walbaum) aquaculture furunculosis outbreaks still occur. In this study we tested the efficacy of experimental subunit vaccines against A. salmonicida infection in rainbow trout. We utilized in silico screening of the proteome of A. salmonicida subsp. salmonicida strain A449 and identified potential protective protein antigens that were tested by in vivo challenge trial. A total of 14 proteins were recombinantly expressed in Escherichia coli and prepared in 3 different subunit vaccine combinations to immunize 3 groups of rainbow trout by intraperitoneal (i.p.) injection. The fish were exposed to virulent A. salmonicida 7 weeks after immunization. To assess the efficacy of the subunit vaccines we evaluated the immune response in fish after immunization and challenge infection by measuring the antibody levels and monitoring the survival of fish in different groups. The survival of fish at 3 weeks after challenge infection showed that all 3 groups of fish immunized with 3 different protein combinations exhibited significantly lower mortalities (17–30%) compared to the control groups (48% and 56%). The ELISA results revealed significantly elevated antibody levels in fish against several protein antigens, which in some cases were positively correlated to the survival. PMID:28182704

  6. Evolutionary and Diagnostic Implications of Intragenomic Heterogeneity in the 16S rRNA Gene in Aeromonas Strains

    PubMed Central

    Morandi, Alessia; Zhaxybayeva, Olga; Gogarten, J. Peter; Graf, Joerg

    2005-01-01

    Sequencing 16S rRNA genes (SSU) cloned from Aeromonas strains revealed that strains contained up to six copies differing by ≤1.5%. The SSU copies from Aeromonas veronii LMG13695 clustered with sequences from four Aeromonas species. These results demonstrate intragenomic heterogeneity of SSU and suggest caution when using SSU to identify aeromonads. PMID:16159790

  7. Biological control of Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) using Aeromonas phage PAS-1.

    PubMed

    Kim, J H; Choresca, C H; Shin, S P; Han, J E; Jun, J W; Park, S C

    2015-02-01

    The potential control efficacy of Aeromonas phage PAS-1 was evaluated against Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) model in this study. The phage was co-cultured with the virulent A. salmonicida subsp. salmonicida strain AS05 that possesses the type III secretion system (TTSS) ascV gene, and efficient bacteriolytic activity was observed against the bacteria. The administration of PAS-1 in rainbow trout demonstrated that the phage was cleared from the fish within 200 h post-administration, and a temporal neutralizing activity against the phage was detected in the sera of phage-administrated fish. The administration of PAS-1 (multiplicity of infection: 10 000) in A. salmonicida subsp. salmonicida infected rainbow trout model showed notable protective effects, with increased survival rates and mean times to death. These results demonstrated that Aeromonas phage PAS-1 could be considered as an alternative biological control agent against A. salmonicida subsp. salmonicida infections in rainbow trout culture.

  8. Production of Non-Ribosomal Peptide Synthetase (NRPS)- Dependent Siderophore by Aeromonas Isolates

    PubMed Central

    Amsaveni, Ramasamy; Sureshkumar, Muthusamy; Aravinth, Arthanari; Mary, Joseph Reshma; Vivekanandhan, Govindasami

    2016-01-01

    Background: Aeromonas species are Gram-negative ubiquitous bacteria, facultative anaerobic rods that infect both invertebrates and vertebrates. Various fish species develop hemorrhagic disease and furunculosis due to Aeromonas spp. Aeromonas strains generate certain active compounds such as siderophores, which are the final products of non-ribosomal peptide synthetase (NRPS) activity. The present study attempted to investigate the prevalence of Aeromonas isolates in marketed fish sources. We also examined the siderophore production ability of these isolates. Methods: Among the molecular tools, 16S rRNA analysis was used to identify Aeromonas species and their epidemiological distributions. The hemolytic activity of the strains and biochemical assays were used to confirm the identity of the isolates. We also determined the chemical nature of siderophores in these strains. Results: A total of seven Aeromonas isolates obtained from fish were included to determine the siderophore production. Of 7 isolates, 4 produced siderophore, and their chemical nature was also determined. The siderophore produced by Aeromonas was invariably found to be of hydroxamate. Four Aeromonas isolates were selected for PCR identification of NRPS-encoding gene. The conserved sequence was present in all four selected isolates. Furthermore, siderophores were qualitatively tested for their antibacterial activity against pathogenic bacteria and a significant level of inhibitory activity was observed in siderophores from the four isolates. Conclusion: Our results showed the ability of the isolated strains in production of siderophores with a high level of activity against Salmonella paratyphi. These siderophores could find applications in biomedical industries. PMID:27155016

  9. Aeromonas caviae strain induces Th1 cytokine response in mouse intestinal tract

    EPA Science Inventory

    Aeromonas caviae has been associated with human gastrointestinal disease. Strains of this species typically lack virulence factors (VFs) such as enterotoxins and hemolysins that are produced by other human pathogens of the Aeromonas genus,. Microarray profiling of...

  10. Aeromonas-Associated Diarrhea in Children Under 5 Years: The GEMS Experience.

    PubMed

    Qamar, Farah Naz; Nisar, Muhammad Imran; Quadri, Farheen; Shakoor, Sadia; Sow, Samba O; Nasrin, Dilruba; Blackwelder, William C; Wu, Yukun; Farag, Tamer; Panchalingham, Sandra; Sur, Dipika; Qureshi, Shahida; Faruque, Abu S G; Saha, Debasish; Alonso, Pedro L; Breiman, Robert F; Bassat, Quique; Tamboura, Boubou; Ramamurthy, Thandavarayan; Kanungo, Suman; Ahmed, Shahnawaz; Hossain, Anowar; Das, Sumon K; Antonio, Martin; Hossain, M Jahangir; Mandomando, Inacio; Tennant, Sharon M; Kotloff, Karen L; Levine, Myron M; Zaidi, Anita K M

    2016-10-05

    We report the clinical findings, epidemiology, and risk factors for moderate-to-severe diarrhea (MSD) associated with Aeromonas species in children 0-59 months of age, from the Global Enteric Multicenter Study, conducted at three sites in south Asia and four sites in sub-Saharan Africa. Children with MSD were enrolled along with controls matched for age, gender, and neighborhood. Pooled, age-stratified conditional logistic regression models were applied to evaluate the association of Aeromonas infection controlling for coinfecting pathogens and sociodemographic variables. A pooled, age-stratified, multivariate logistic regression analysis was done to identify risk factors associated with Aeromonas positivity in MSD cases. A total of 12,110 cases and 17,291 matched controls were enrolled over a period of 48 months. Aeromonas was identified as a significant pathogen in 736 cases of MSD in Pakistan and Bangladesh (22.2%). Aeromonas remained a significant pathogen even after adjustment for the presence of other pathogens and sociodemographic factors. Odds ratio (OR) for Aeromonas were higher in the presence of Shigella (matched OR: 6.2, 95% confidence interval [CI]: 1.9-20.2). Cases of Aeromonas were likely to present with dysentery, particularly in the 0-11 months (OR: 1.4, 95% CI 1.0-2.0) and 12-23 months (OR: 1.8, 95% CI: 1.3-2.5) age group. The odds of Aeromonas increased with increasing degree of stunting, being highest for severe stunting (OR: 10.1, 95% CI: 3.6-28.9). Aeromonas is a significant pathogen for MSD in Pakistan and Bangladesh. Presence of dysentery and co-occurrence with other pathogens, notably Shigella spp. are significant features of Aeromonas-associated diarrhea.

  11. Effect of solar irradiation on extracellular enzymes of Aeromonas proteolytica

    NASA Technical Reports Server (NTRS)

    Foster, B. G.

    1973-01-01

    The bacterium Aeromonas proteolytica was selected for studying the effects of solar irradiation on extracellular enzymes because it produces an endopeptidase that is capable of degrading proteins and a hemolysin that is active in lysing human erythrocytes. Possible alterations in the rate of enzyme production in response to the test conditions are currently underway and are not available for this preliminary report. Completed viability studies are indicative that little difference exists among the survival curves derived for cells exposed to various components of ultraviolet irradiation in space.

  12. Differences in production of several extracellular virulence factors in clinical and food Aeromonas spp. strains.

    PubMed

    Pin, C; Marín, M L; Selgas, D; García, M L; Tormo, J; Casas, C

    1995-02-01

    Production of several extracellular virulence factors (lipase, protease and haemolysin) was compared in 15 Aeromonas spp. isolated from faeces of patients with Aeromonas-associated gastroenteritis and 81 strains isolated from food. Strains from food did not show differences in production of these factors when compared with strains isolated from faeces. However, if strains were considered in relation to autoagglutination (AA) character, the AA+ differed from AA- strains in lipase and protease production. Supernatant fluids of AA+ food and human strains showed 2.5-fold more protease production than that observed in AA- strains. These two characteristics of certain Aeromonas strains could be related with the more virulent capacity.

  13. Antibiotic resistance in Aeromonas upstream and downstream of a water resource recovery facility.

    PubMed

    Cisar, Cindy R; Henderson, Samantha K; Askew, Maegan L; Risenhoover, Hollie G; McAndrews, Chrystle R; Kennedy, S Dawn; Paine, C Sue

    2014-09-01

    Aeromonas strains isolated from sediments upstream and downstream of a water resource recovery facility (WRRF) over a two-year time period were tested for susceptibility to 13 antibiotics. Incidence of resistance to antibiotics, antibiotic resistance phenotypes, and diversity (based on resistance phenotypes) were compared in the two populations. At the beginning of the study, the upstream and downstream Aeromonas populations were different for incidence of antibiotic resistance (p < 0.01), resistance phenotypes (p < 0.005), and diversity. However, these differences declined over time and were not significant at the end of the study. These results (1) indicate that antibiotic resistance in Aeromonas in stream sediments fluctuates considerably over time and (2) suggest that WRRF effluent does not, when examined over the long- term, affect antibiotic resistance in Aeromonas in downstream sediment.

  14. IDENTIFICATION AND CHARACTERIZATION OF AEROMONAS ISOLATES FROM DRINKING WATER DISTRIBUTION SYSTEMS

    EPA Science Inventory

    Members of the bacterial genus Aeromonas are commonly isolated from both fresh and salt waters worldwide and some are believed to cause infections in humans, including gastroenteritis and wound infections. Currently, aeromonads are on the United States Environmental Protection A...

  15. Escherichia Coli

    ERIC Educational Resources Information Center

    Goodsell, David S.

    2009-01-01

    Diverse biological data may be used to create illustrations of molecules in their cellular context. I describe the scientific results that support a recent textbook illustration of an "Escherichia coli cell". The image magnifies a portion of the bacterium at one million times, showing the location and form of individual macromolecules. Results…

  16. Molecular Detection, Quantification, and Toxigenicity Profiling of Aeromonas spp. in Source- and Drinking-Water.

    PubMed

    Robertson, Boakai K; Harden, Carol; Selvaraju, Suresh B; Pradhan, Suman; Yadav, Jagjit S

    2014-01-01

    Aeromonas is ubiquitous in aquatic environments and has been associated with a number of extra-gastrointestinal and gastrointestinal illnesses. This warrants monitoring of raw and processed water sources for pathogenic and toxigenic species of this human pathogen. In this study, a total of 17 different water samples [9 raw and 8 treated samples including 4 basin water (partial sand filtration) and 4 finished water samples] were screened for Aeromonas using selective culturing and a genus-specific real-time quantitative PCR assay. The selective culturing yielded Aeromonas counts ranging 0 - 2 x 10(3)CFU/ml and 15 Aeromonas isolates from both raw and treated water samples. The qPCR analysis indicated presence of a considerable nonculturable population (3.4 x 10(1) - 2.4 x 10(4) cells/ml) of Aeromonas in drinking water samples. Virulence potential of the Aeromonas isolates was assessed by multiplex/singleplex PCR-based profiling of the hemolysin and enterotoxin genes viz cytotoxic heat-labile enterotoxin (act), heat-labile cytotonic enterotoxin (alt), heat-stable cytotonic enterotoxin (ast), and aerolysin (aerA) genes. The water isolates yielded five distinct toxigenicity profiles, viz. act, alt, act+alt, aerA+alt, and aerA+alt+act. The alt gene showed the highest frequency of occurrence (40%), followed by the aerA (20%), act (13%), and ast (0%) genes. Taken together, the study demonstrated the occurrence of a considerable population of nonculturable Aeromonads in water and prevalence of toxigenic Aeromonas spp. potentially pathogenic to humans. This emphasizes the importance of routine monitoring of both source and drinking water for this human pathogen and role of the developed molecular approaches in improving the Aeromonas monitoring scheme for water.

  17. Gastroenteritis caused by Aeromonas trota in a child.

    PubMed Central

    Reina, J; Lopez, A

    1996-01-01

    A case of acute diarrhoea caused by Aeromonas trota (formerly HG 13 group) in a Spanish child is reported. The strain was isolated in the faeces using the CIN agar (cefsulodin-irgasan-novobiocin) culture media. The strain was initially identified as A sobria by the commercial GNI card and API 20E biochemical systems. The strain was, however, VogesProskauer and sucrose negative, so complementary tests of cellobiose fermentation and gluconate oxidation were performed. These tests, together with the strain susceptibility to ampicillin (MIC 1 microgram/ml) and carbenicillin (MIC < 16 micrograms/ml) led to the final identification of A trota. The microbiological characteristics of this new species and the principal tests required for its identification are presented. The isolation, for the first time, of A trota in the Mediterranean area confirms the suspected worldwide distribution of this species. PMID:8655689

  18. Occurrence of motile Aeromonas in municipal drinking water and distribution of genes encoding virulence factors.

    PubMed

    Pablos, Manuel; Rodríguez-Calleja, Jose M; Santos, Jesús A; Otero, Andrés; García-López, María-Luisa

    2009-10-31

    Aeromonas-associated cases of gastroenteritis are generally considered waterborne. The purpose of this study was to evaluate the potential microbiological risk associated with the presence of these bacteria in public drinking water. Over a period of one year, 132 drinking-water samples were monitored in León (NW of Spain, 137,000 inhabitants) for mandatory drinking-water standards and the occurrence of Aeromonas spp. Samples were taken at the municipal water treatment plant, one storage facility, and two public artesian drinking-water fountains. Because of low numbers of coliforms or Clostridium perfringens, the non-compliance rate with microbial standards was 3.8% whereas the percentage of positive samples for motile mesophilic Aeromonas was 26.5%. For all but two samples, Aeromonas was recovered between October and early March when the temperature was below 14 degrees C and the residual chlorine ranged from 0.21 to 0.72 mg/l. An apparent relationship was observed between rainfall and the incidence of Aeromonas. The 35 selected Aeromonas isolates were identified as A. caviae and A. media. The alt and laf genes were present in all isolates, the aerA gene was present in six isolates, and the four remaining genes investigated (hlyA, ast, stx1 and stx2) were absent. The combinations of putative virulence genes were: aerA(-)/hlyA(-)/alt(+)/ast(-)/laf(+)/stx1(-)/stx2(-) (82.9%) and aerA(+)/hlyA(-)/alt(+)/ast(-)/laf(+)/stx1(-)/stx2(-) (17.1%). None of the isolates bore plasmids. As Aeromonas strains harbouring two or more virulence-associated genes have the potential to cause disease by direct transmission via drinking water or by water use in food preparation, it would be advisable to control excessive numbers of these bacteria in drinking-water supplies.

  19. Low Prevalence of Aeromonas hydrophilain Infectious Diarrhea Samples of Pediatric Patients in Arak, Iran

    PubMed Central

    Elnaz, Abbasi; Behzad, Khansari-nejad; Hamid, Abtahi; Majid, Akbari; Ehsanollah, Ghaznavi-rad

    2016-01-01

    Background: Aeromonashydrophila (A. hydrophila) is an aquatic bacterium that can cause a spectrum of infectious diseases, including both gastrointestinal and extraintestinal infections. Due to the high rate of diarrheal infections in pediatric patients in central Iran, this study was designed to determine the frequency of A. hydrophila in diarrhea samples from children in this region. Methods: In this descriptive cross-sectional study, diarrheal stool specimens were collected from 200 children admitted between February and October of 2015 to educational and medical centers affiliated with the Arak University of Medical Sciences. The samples were analyzed both phenotypically by culture and genotypically by the polymerase chain reaction (PCR). Results: A. hydrophila was isolated from two of the 200 samples tested (1%). The presence of bacterial genetic markers further confirmed the diagnosis. Conclusion: Based on this study, A. hydrophilais not highly prevalent in children with diarrhea in Arak; however clinical diagnostic laboratory personnel should be aware of the possible presence of A.hydrophila in children with diarrhea as it can cause dangerous health problems in both them and young adolescents. PMID:28070529

  20. Identification and Molecular Characterization of the Homogentisate Pathway Responsible for Pyomelanin Production, the Major Melanin Constituents in Aeromonas media WS

    PubMed Central

    Wang, He; Qiao, Yunqian; Chai, Baozhong; Qiu, Chenxi; Chen, Xiangdong

    2015-01-01

    The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine) based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate). Gene products of phhA (encodes phenylalanine hydroxylase), tyrB and aspC (both encode aromatic amino acid aminotransferase), and hppD (encodes 4-hydroxyphenylpyruvate dioxygenase) constitute a linear pathway of converting phenylalanine to HGA and disruption of any one of these genes impairs or blocks pigmentation of A. media strain WS. This HGA biosynthesis pathway is widely distributed in Aeromonas, but HGA is only detectable in the cultures of pigmented Aeromonas species. Heterologous expression of HppD from both pigmented and non-pigmented Aeromonas species in E. coli leads to the production of pyomelanin and thus pigmentation, suggesting that most Aeromonas species have the critical enzymes to produce pyomelanin through HGA. Taken together, we have identified a widely conserved biosynthesis pathway of HGA based pyomelanin in Aeromonas that may be responsible for pigmentation of many Aeromonas species. PMID:25793756

  1. A prospective study on Aeromonas in outpatients with diarrhea in the central region of Rio Grande do Sul State

    PubMed Central

    Prediger, Karoline deCampos; Pereira, Renata da Silva; Winckler Neto, Carlos Hugo Del Priore; Santos, Roberto Christ Vianna; Fadel-Picheth, Cyntia Maria Telles; Vizzotto, Bruno Stefanello

    2012-01-01

    Aeromonas spp. were identified in five (2,7%) of 182 diarrheal stool cultures, A. caviae was predominant, resistant mainly to ampicillin and cephalotin. This is the first study showing the presence of Aeromonas spp. in diarrheal stools of outpatients in the central region of Rio Grande do Sul State, Brazil. PMID:24031914

  2. Isolation and partial characterization of a virulent bacteriophage IHQ1 specific for Aeromonas punctata from stream water.

    PubMed

    Ul Haq, Irshad; Chaudhry, Waqas Nasir; Andleeb, Saadia; Qadri, Ishtiaq

    2012-05-01

    Aeromonas punctata is the causative agent of septicemia, diarrhea, wound infections, meningitis, peritonitis, and infections of the joints, bones and eyes. Bacteriophages are often considered alternative agents for controlling bacterial infection and contamination. In this study, we described the isolation and preliminary characterization of bacteriophage IHQ1 (family Myoviridae) active against the Gram-negative bacterial strain A. punctata. This virulent bacteriophage was isolated from stream water sample. Genome analysis indicated that phage IHQ1 was a double-stranded DNA virus with an approximate genome size of 25-28 kb. The initial characterization of this newly isolated phage showed that it has a narrow host range and infects only A. punctata as it failed to infect seven other clinically isolated pathogenic strains, i.e., methicillin-resistant Staphylococcus aureus 6403, MRSA 17644, Acinetobacter 33408, Acinetobacter 1172, Pseudomonas aeruginosa 22250, P. aeruginosa 11219, and Escherichia coli. Proteomic pattern of phage IHQ1, generated by SDS-PAGE using purified phage particles, showed three major and three minor protein bands with molecular weights ranging from 25 to 70 kDa. The adsorption rate of phage IHQ1 to the host bacterium was also determined, which was significantly enhanced by the addition of 10 mM CaCl(2). From the single-step growth experiment, it was inferred that the latent time period of phage IHQ1 was 24 min and a burst size of 626 phages per cell. Moreover, the pH and thermal stability of phage IHQ1 were also investigated. The maximum stability of the phage was observed at optimal pH 7.0, and it was totally unstable at extreme acidic pH 3; however, it was comparatively stable at alkaline pH 11.0. At 37°C the phage showed maximum number of plaques, and the viability was almost 100%. The existence of Aeromonas bacteriophage is very promising for the eradication of this opportunistic pathogen and also for future applications such as the

  3. A Molecular Study on the Prevalence and Virulence Potential of Aeromonas spp. Recovered from Patients Suffering from Diarrhea in Israel

    PubMed Central

    Senderovich, Yigal; Ken-Dror, Shifra; Vainblat, Irina; Blau, Dvora; Izhaki, Ido; Halpern, Malka

    2012-01-01

    Background Species of the genus Aeromonas are native inhabitants of aquatic environments and have recently been considered emerging human pathogens. Although the gastrointestinal tract is by far the most common anatomic site from which aeromonads are recovered, their role as etiologic agents of bacterial diarrhea is still disputed. Aeromonas-associated diarrhea is a phenomenon occurring worldwide; however, the exact prevalence of Aeromonas infections on a global scale is unknown. Methodology/Principal Findings The prevalence and virulence potential of Aeromonas in patients suffering from diarrhea in Israel was studied using molecular methods. 1,033 diarrheal stools were sampled between April and September 2010 and Aeromonas species were identified in 17 (∼2%) patients by sequencing the rpoD gene. Aeromonas species identity and abundance was: A. caviae (65%), A. veronii (29%) and Aeromonas taiwanensis (6%). This is the first clinical record of A. taiwanensis as a diarrheal causative since its recent discovery from a wound infection in a patient in Taiwan. Most of the patients (77%) from which Aeromonas species were isolated were negative for any other pathogens. The patients ranged from 1 to 92 years in age. Aeromonas isolates were found to possess different virulence-associated genes: ahpB (88%), pla/lip/lipH3/apl-1 (71%), act/hlyA/aerA (35%), alt (18%), ast (6%), fla (65%), lafA (41%), TTSS ascV (12%), TTSS ascF-ascG (12%), TTSS-dependent ADP-ribosylating toxins aexU (41%) and aexT (6%) in various combinations. Most of the identified strains were resistant to beta-lactam antibiotics but susceptible to third-generation cephalosporin antibiotics. Conclusions Aeromonas may be a causative agent of diarrhea in patients in Israel and therefore should be included in routine bacteriological screenings. PMID:22355306

  4. Maturation pathway of metalloprotease produced by Aeromonas sobria.

    PubMed

    Takahashi, Eizo; Fujii, Yoshio; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Nair, Gopinath Balakrish; Takeda, Yoshifumi; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

    2010-10-01

    Previously, we cloned the metalloprotease gene of Aeromonas sobria (amp) and determined its nucleotide sequence (GenBank accession number DQ784565). The protease is composed of 591 amino acid residues. In this study, we purified the mature metalloprotease from the culture supernatant of A. sobria and determined the amino terminal sequence and molecular size of AMP. In addition, we examined the production of AMP diachronically and found that AMP emerges outside of the cell as an intermediate composed of mature and propeptide regions. Subsequently, we determined that the N-terminal amino acid sequence of the intermediate and found that the sequence is identical to that of the mature metalloprotease. This means that the intermediate is composed of a mature AMP region and a C-terminal propeptide. The cross culture experiment of mutants of metalloprotease and serine protease of A. sobria on skim milk agar medium indicates that the intermediate released outside of the cell is inactive and that serine protease produced by A. sobria accelerates the conversion of the intermediate from the inactive to the active form.

  5. Lectin-binding properties of Aeromonas caviae strains

    PubMed Central

    Rocha-de-Souza, Cláudio M.; Hirata-Jr, Raphael; Mattos-Guaraldi, Ana L.; Freitas-Almeida, Angela C.; Andrade, Arnaldo F. B.

    2008-01-01

    The cell surface carbohydrates of four strains of Aeromonas caviae were analyzed by agglutination and lectin-binding assays employing twenty highly purified lectins encompassing all sugar specificities. With the exception of L-fucose and sialic acid, the sugar residues were detected in A. caviae strains. A marked difference, however, in the pattern of cell surface carbohydrates in different A. caviae isolates was observed. Specific receptors for Tritricum vulgaris (WGA), Lycopersicon esculentum (LEL) and Solanum tuberosum (STA) (D-GlcNAc-binding lectins) were found only in ATCC 15468 strain, whereas Euonymus europaeus (EEL, D-Gal-binding lectin) sites were present exclusively in AeQ32 strain, those for Helix pomatia (HPA, D-GalNAc-binding lectin) in AeC398 and AeV11 strains, and for Canavalia ensiformes (Con A, D-Man-binding lectin) in ATCC 15468, AeC398, AeQ32 and AeV11 strains, after bacterial growing at 37°C. On the other hand, specific receptors for WGA and EEL were completely abrogated growing the bacteria at 22°C. Binding studies with 125I- labeled lectins from WGA, EEL and Con A were performed. These assays essentially confirmed the selectivity, demonstrated in the agglutination assays of these lectins for the A. caviae strains. PMID:24031204

  6. Levels of enzymes in leukaemic mice treated withAeromonas L-asparaginase.

    PubMed

    Benny, P J; Muraleedhara Kurup, G; Sreejith, K

    1999-07-01

    L-asparaginase isolated in our laboratory fromAeromonas has been found to be antileukaemic. In the present study changes in the levels of serum enzymes in leukaemic mice and under treatment withAeromonas L-asparaginase has been compared. A significant increase in the levels of serum lactate dehydrogenase with tumour growth and a decrease during therapy was observed. A significant decrease in alanine transaminase activity during tumour growth and an increase during treatment was noticed. Increased levels of aspartate transaminase and alkaline phosphatase was observed during enzyme therapy. Total acid phosphatase was found to be increased during tumour growth and decreased considerably during treatment.

  7. Metabolism of proteins and glycoproteins in tumour bearing mice treated with Aeromonas L-asparaginase.

    PubMed

    Benny, P J; Muraleedhara, K G; Sreejith, K; Jayashree, G

    1996-12-01

    L-asparaginase, isolated in our laboratory, from Aeromonas had been found to be antileukaemic. In the present study, changes in the levels of proteins and glycoproteins in leukaemic mice and under treatment with Aeromonas L-asparaginase have been compared. Levels of protein bound hexose, fucose and sialic acid which were increased during leukaemia attained normal levels when treated with L-asparaginase. The increased blood urea level declined significantly during enzyme therapy. Effects of L-asparaginase are compared with 'Leunase', a commercially available drug used in the treatment of leukaemia.

  8. Integrated Bioenvironmental Hazards Research Program

    DTIC Science & Technology

    2004-01-01

    Aeromonas hydrophila, Chromobacterium violaceum , and Burkholderia cepacia strains isolated from different sites and at different times of year...indicated that the major genera of these bacteria were Aeromonas, Chromobacterium , Enterobacter, Burkholderia, and Pseudomonas. The majority of the...Spatial and seasonal conditions had no significant effect on the antibiotic resistance patterns of Aeromonas hydrophila and Chromabacterium violaceum

  9. Molecular characterization of Shewanella and Aeromonas isolates associated with spoilage of Common carp (Cyprinus carpio).

    PubMed

    Beaz-Hidalgo, Roxana; Agüeria, Daniela; Latif-Eugenín, Fadua; Yeannes, Maria I; Figueras, Maria J

    2015-01-01

    Storage in ice is a common way of preserving commercial fish species but some microorganisms can still contaminate and participate in the spoilage of the product; therefore, identification of potential harmful microbes is important. Thirteen colonies were isolated from common carp (Cyprinus carpio) that had been stored in ice, whose phenotypic identification revealed that they belonged to the genera Aeromonas (n = 5) and Shewanella (n = 8). Molecular genotyping with ERIC-PCR showed clonality only among two of the five Aeromonas isolates and for two groups (n = 3; n = 2) of the eight Shewanella isolates. Sequencing the rpoD gene showed that four Aeromonas isolates belonged to the species Aeromonas salmonicida and one to A. sobria. Of the eight Shewanella, seven isolates cluster with Shewanella putrefaciens and one with Shewanella profunda in the 16S rRNA phylogenetic tree. However, analysis of the gyrB gene showed that these eight isolates could constitute a new species closely related to S. baltica. The Shewanella and A. salmonicida isolates produce off-odours and reduce trimethylamine oxide, indicating that they might contribute to the spoilage of the fish.

  10. Draft Genome Sequence of Marine-Derived Aeromonas caviae CHZ306, a Potential Chitinase Producer Strain

    PubMed Central

    Zimpel, Cristina Kraemer; Guimaraes, Ana Marcia Sa; Pessoa, Adalberto; Rivera, Irma Nelly Gutierrez

    2016-01-01

    We report here a draft genome sequence of Aeromonas caviae CHZ306, a marine-derived bacterium with the ability to hydrolyze chitin and express high levels of chitinases. The assembly resulted in 65 scaffolds with approximately 4.78 Mb. Genomic analysis revealed different genes encoding chitin-degrading enzymes that can be used for chitin derivative production. PMID:27856589

  11. Diversity and antibiotic resistance of Aeromonas spp. in drinking and waste water treatment plants.

    PubMed

    Figueira, Vânia; Vaz-Moreira, Ivone; Silva, Márcia; Manaia, Célia M

    2011-11-01

    The taxonomic diversity and antibiotic resistance phenotypes of aeromonads were examined in samples from drinking and waste water treatment plants (surface, ground and disinfected water in a drinking water treatment plant, and raw and treated waste water) and tap water. Bacteria identification and intra-species variation were determined based on the analysis of the 16S rRNA, gyrB and cpn60 gene sequences. Resistance phenotypes were determined using the disc diffusion method. Aeromonas veronii prevailed in raw surface water, Aeromonas hydrophyla in ozonated water, and Aeromonas media and Aeromonas puntacta in waste water. No aeromonads were detected in ground water, after the chlorination tank or in tap water. Resistance to ceftazidime or meropenem was detected in isolates from the drinking water treatment plant and waste water isolates were intrinsically resistant to nalidixic acid. Most of the times, quinolone resistance was associated with the gyrA mutation in serine 83. The gene qnrS, but not the genes qnrA, B, C, D or qepA, was detected in both surface and waste water isolates. The gene aac(6')-ib-cr was detected in different waste water strains isolated in the presence of ciprofloxacin. Both quinolone resistance genes were detected only in the species A. media. This is the first study tracking antimicrobial resistance in aeromonads in drinking, tap and waste water and the importance of these bacteria as vectors of resistance in aquatic environments is discussed.

  12. Draft Genome Sequence of Marine-Derived Aeromonas caviae CHZ306, a Potential Chitinase Producer Strain.

    PubMed

    Cardozo, Flávio Augusto; Zimpel, Cristina Kraemer; Guimaraes, Ana Marcia Sa; Pessoa, Adalberto; Rivera, Irma Nelly Gutierrez

    2016-11-17

    We report here a draft genome sequence of Aeromonas caviae CHZ306, a marine-derived bacterium with the ability to hydrolyze chitin and express high levels of chitinases. The assembly resulted in 65 scaffolds with approximately 4.78 Mb. Genomic analysis revealed different genes encoding chitin-degrading enzymes that can be used for chitin derivative production.

  13. VIRULENCE FACTORS OF AEROMONAS: A GENETIC CHARACTERIZATION OF DRINKING WATER ISOLATES

    EPA Science Inventory

    A survey of finished drinking water conducted by the US EPA during 2000-2001, revealed that 8 out of 18 water utilities encompassing several states (NY, KY, IA, OH) were contaminated with aeromonas species. Altogether 205 organisms were isolated by EPA method 1601. All of the ...

  14. A Mouse Model for Characterization of Gastrointestinal Colonization Rates Among Environmental Aeromonas Isolates

    EPA Science Inventory

    The colonization rates of ten different environmental isolates of Aeromonas were determined using a novel mouse-streptomycin pre-treatment method. A novel streptomycin pre-treatment prepared animals with a transient alteration in colon flora that allowed colonization by Aeromon...

  15. Lippia alba essential oil promotes survival of silver catfish (Rhamdia quelen) infected with Aeromonas sp.

    PubMed

    Sutili, Fernando J; Cunha, Mauro A; Ziech, Rosangela E; Krewer, Carina C; Zeppenfeld, Carla C; Heldwein, Clarissa G; Gressler, Leticia T; Heinzmann, Berta M; Vargas, Agueda C; Baldisserotto, Bernardo

    2015-03-01

    In vitro and in vivo activity of the Lippia alba essential oil (EO) against Aeromonas sp. was evaluated. In the in vitro assay the minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of EO for Aeromonas cells were determined using the microdilution method. Twenty five strains of Aeromonas sp. isolated from infected fish obtained from local fish farms were used. MIC and MBC values were 2862 and 5998 µg mL-1 for L. alba EO and 0.5 and 1.2 µg mL-1 for gentamicin, respectively. In the in vivo assay silver catfish juveniles (Rhamdia quelen) (7.50 ± 1.85 g and 10.0 ± 1.0 cm) with typical injuries associated to Aeromonas infection were divided into four treatments (in triplicate n=10): untreated fish (negative control), 10 mg L-1 of gentamicin, and 20 or 50 µL L-1 of EO. Fish were maintained in aerated 20 L plastic boxes. After 10 days survival of silver catfish infected with Aermonas sp. and treated with essential oil (50 µL L-1) was greater than 90%.

  16. Antibiotic susceptibility profile of Aeromonas species isolated from wastewater treatment plant.

    PubMed

    Igbinosa, Isoken H; Okoh, Anthony I

    2012-01-01

    This study assessed the prevalence of antibiotic-resistant Aeromonas species isolated from Alice and Fort Beaufort wastewater treatment plant in the Eastern Cape Province of South Africa. Antibiotic susceptibility was determined using the disc diffusion method, and polymerase chain reaction (PCR) assay was employed for the detection of antibiotics resistance genes. Variable susceptibilities were observed against ciprofloxacin, chloramphenicol, nalidixic acid, gentamicin, minocycline, among others. Aeromonas isolates from both locations were 100% resistant to penicillin, oxacillin, ampicillin, and vancomycin. Higher phenotypic resistance was observed in isolates from Fort Beaufort compared to isolates from Alice. Class A pse1 β-lactamase was detected in 20.8% of the isolates with a lower detection rate of 8.3% for bla(TEM) gene. Class 1 integron was present in 20.8% of Aeromonas isolates while class 2 integron and TetC gene were not detected in any isolate. The antibiotic resistance phenotypes observed in the isolates and the presence of β-lactamases genes detected in some isolates are of clinical and public health concern as this has consequences for antimicrobial chemotherapy of infections associated with Aeromonas species. This study further supports wastewater as potential reservoirs of antibiotic resistance determinants in the environment.

  17. The mosaic architecture of Aeromonas salmonicida subsp. salmonicida pAsa4 plasmid and its consequences on antibiotic resistance

    PubMed Central

    Tanaka, Katherine H.; Vincent, Antony T.; Trudel, Mélanie V.; Paquet, Valérie E.; Frenette, Michel

    2016-01-01

    Aeromonas salmonicida subsp. salmonicida, the causative agent of furunculosis in salmonids, is an issue especially because many isolates of this bacterium display antibiotic resistances, which limit treatments against the disease. Recent results suggested the possible existence of alternative forms of pAsa4, a large plasmid found in A. salmonicida subsp. salmonicida and bearing multiple antibiotic resistance genes. The present study reveals the existence of two newly detected pAsa4 variants, pAsa4b and pAsa4c. We present the extensive characterization of the genomic architecture, the mobile genetic elements and the antimicrobial resistance genes of these plasmids in addition to the reference pAsa4 from the strain A449. The analysis showed differences between the three architectures with consequences on the content of resistance genes. The genomic plasticity of the three pAsa4 variants could be partially explained by the action of mobile genetic elements like insertion sequences. Eight additional isolates from Canada and Europe that bore similar antibiotic resistance patterns as pAsa4-bearing strains were genotyped and specific pAsa4 variants could be attributed to phenotypic profiles. pAsa4 and pAsa4c were found in Europe, while pAsa4b was found in Canada. In accordance with their content in conjugative transfer genes, only pAsa4b and pAsa4c can be transferred by conjugation in Escherichia coli. The plasticity of pAsa4 variants related to the acquisition of antibiotic resistance indicates that these plasmids may pose a threat in terms of the dissemination of antimicrobial-resistant A. salmonicida subsp. salmonicida bacteria. PMID:27812409

  18. Effect of temperature on growth and activity of Aeromonas spp. and mixed bacterial populations in the Anacostia River.

    PubMed Central

    Cavari, B Z; Allen, D A; Colwell, R R

    1981-01-01

    During the winter months, total bacterial counts in the water column and in the sediment in the Anacostia River were two- to eightfold higher than at other times of the year, whereas Aeromonas spp. decreased in number of several orders of magnitude. This significant decrease in number in the Anacostia River during the cold months of the year can be explained by the low metabolic activity of Aeromonas at low temperatures. PMID:7235703

  19. Effect of temperature on growth and activity of Aeromonas spp. and mixed bacterial populations in the Anacostia River.

    PubMed

    Cavari, B Z; Allen, D A; Colwell, R R

    1981-04-01

    During the winter months, total bacterial counts in the water column and in the sediment in the Anacostia River were two- to eightfold higher than at other times of the year, whereas Aeromonas spp. decreased in number of several orders of magnitude. This significant decrease in number in the Anacostia River during the cold months of the year can be explained by the low metabolic activity of Aeromonas at low temperatures.

  20. Reciprocal immune benefit based on complementary production of antibiotics by the leech Hirudo verbana and its gut symbiont Aeromonas veronii

    PubMed Central

    Tasiemski, Aurélie; Massol, François; Cuvillier-Hot, Virginie; Boidin-Wichlacz, Céline; Roger, Emmanuel; Rodet, Franck; Fournier, Isabelle; Thomas, Frédéric; Salzet, Michel

    2015-01-01

    The medicinal leech has established a long-term mutualistic association with Aeromonas veronii, a versatile bacterium which can also display free-living waterborne and fish- or human-pathogenic lifestyles. Here, we investigated the role of antibiotics in the dynamics of interaction between the leech and its gut symbiont Aeromonas. By combining biochemical and molecular approaches, we isolated and identified for the first time the antimicrobial peptides (AMPs) produced by the leech digestive tract and by its symbiont Aeromonas. Immunohistochemistry data and PCR analyses evidenced that leech AMP genes are induced in the gut epithelial cells when Aeromonas load is low (starved animals), while repressed when Aeromonas abundance is the highest (post blood feeding). The asynchronous production of AMPs by both partners suggests that these antibiotic substances (i) provide them with reciprocal protection against invasive bacteria and (ii) contribute to the unusual simplicity of the gut microflora of the leech. This immune benefit substantially reinforces the evidence of an evolutionarily stable association between H. verbana and A. veronii. Altogether these data may provide insights into the processes making the association with an Aeromonas species in the digestive tract either deleterious or beneficial. PMID:26635240

  1. Reciprocal immune benefit based on complementary production of antibiotics by the leech Hirudo verbana and its gut symbiont Aeromonas veronii.

    PubMed

    Tasiemski, Aurélie; Massol, François; Cuvillier-Hot, Virginie; Boidin-Wichlacz, Céline; Roger, Emmanuel; Rodet, Franck; Fournier, Isabelle; Thomas, Frédéric; Salzet, Michel

    2015-12-04

    The medicinal leech has established a long-term mutualistic association with Aeromonas veronii, a versatile bacterium which can also display free-living waterborne and fish- or human-pathogenic lifestyles. Here, we investigated the role of antibiotics in the dynamics of interaction between the leech and its gut symbiont Aeromonas. By combining biochemical and molecular approaches, we isolated and identified for the first time the antimicrobial peptides (AMPs) produced by the leech digestive tract and by its symbiont Aeromonas. Immunohistochemistry data and PCR analyses evidenced that leech AMP genes are induced in the gut epithelial cells when Aeromonas load is low (starved animals), while repressed when Aeromonas abundance is the highest (post blood feeding). The asynchronous production of AMPs by both partners suggests that these antibiotic substances (i) provide them with reciprocal protection against invasive bacteria and (ii) contribute to the unusual simplicity of the gut microflora of the leech. This immune benefit substantially reinforces the evidence of an evolutionarily stable association between H. verbana and A. veronii. Altogether these data may provide insights into the processes making the association with an Aeromonas species in the digestive tract either deleterious or beneficial.

  2. EVALUATING VIRULENCE OF WATERBORNE AND CLINCIAL AEROMONAS ISOLATES USING GENE EXPRESSION AND MORTALITY IN NEONATAL MICE FOLLOWED BY ASSESSING CELL CULTURE'S ABILITY TO PREDICT VIRULENCE BASED ON TRANSCRIPTIONAL RESPONSE

    EPA Science Inventory

    The virulence of multiple Aeromonas spp. were assessed using two models, a neonatal mouse assay and a mouse intestinal cell culture. Transcriptional responses to both infection models were assessed using microarrays. After artificial infection with a variety of Aeromonas spp., ...

  3. Lipid metabolism in tumour bearing mice treated withAeromonas L-asparaginase.

    PubMed

    Benny, P J; Kurup, G M; Sreejith, K

    1997-07-01

    The anticancerous drug isolated in our laboratory from estuarineAeromonas was characterised and is found to be an enzyme, L-asparaginase. The antileukaemic effect of this drug was studied in mice by inducing leukaemia with Ehrlich ascites cell lines. It was compared with commercially available drug, Leunase, isolated fromE. coli. The lipid profiles in mice during leukaemia and under treatment was studied. The decreased levels of cholesterol and increased levels of triglycerides and phospholipids in serum, liver and kidney were observed in tumour bearing mice. Significant changes in the above values were observed with enzyme therapy. It could bring some of the values to near normal level. L-asparaginase fromAeromonas was found to be more effective.

  4. The Social Life of Aeromonas through Biofilm and Quorum Sensing Systems

    PubMed Central

    Talagrand-Reboul, Emilie; Jumas-Bilak, Estelle; Lamy, Brigitte

    2017-01-01

    Bacteria of the genus Aeromonas display multicellular behaviors herein referred to as “social life”. Since the 1990s, interest has grown in cell-to-cell communication through quorum sensing signals and biofilm formation. As they are interconnected, these two self-organizing systems deserve to be considered together for a fresh perspective on the natural history and lifestyles of aeromonads. In this review, we focus on the multicellular behaviors of Aeromonas, i.e., its social life. First, we review and discuss the available knowledge at the molecular and cellular levels for biofilm and quorum sensing. We then discuss the complex, subtle, and nested interconnections between the two systems. Finally, we focus on the aeromonad multicellular coordinated behaviors involved in heterotrophy and virulence that represent technological opportunities and applied research challenges. PMID:28163702

  5. Aeromonas caviae strain induces Th1 cytokine response in mouse intestinal tract

    SciTech Connect

    Hayes, S L; Lye, D J; McKinstry, Craig A.; Vesper, Sephen J.

    2010-01-01

    Aeromonas caviae has been associated with human gastrointestinal disease. Strains of this species typically lack virulence factors (VFs) such as enterotoxins and hemolysins that are produced by other human pathogens of the Aeromonas genus. Microarray profiling of murine small intestinal extracts, 24 hours after oral infection with an A. caviae strain, provides evidence of a Th1 type immune response. A large number of gamma-interferon (γ-IFN) induced genes are up-regulated as well as several tumor necrosis factor-alpha (TNF-α) transcripts. A. caviae has always been considered as opportunistic pathogen because it lacks obvious virulence factors. This current effort suggests that an A. caviae strain can colonize the murine intestinal tract and cause what has been described by others as a dysregulatory cytokine response. This response could explain why a number of diarrheal waterborne disease cases have been attributed to A. caviae even though it lacks obvious enteropathogenic properties.

  6. A comparison of different culture media for the membrane filter quantification of Aeromonas in water.

    PubMed

    Villari, P; Pucino, A; Santagata, N; Torre, I

    1999-10-01

    A comparative assessment of culture media for the membrane filter enumeration of Aeromonas spp. in water was performed, testing the effects of different incubation conditions (aerobic and anaerobic), temperatures (30 and 37 degrees C) and times (24 and 48 h). Different water samples seeded with test suspensions of Aeromonas spp., fecal material or raw sewage were examined. Results indicate clearly that plates should be incubated aerobically at 30 degrees C for 24 h. If the bacterial contamination is likely to be low, the use of most sensitive culture media, such as SAA, mA, ADA or PADE Agar, is recommended. By contrast, samples with an expected high level of background microbial flora should be analysed through more selective media, such as MIX Agar. However, the low selectivity of all media tested and the high likelihood of false negatives based upon the macroscopic examination of colonies means that further research directed to the development of more efficient media is needed.

  7. Bioinformatic Genome Comparisons for Taxonomic and Phylogenetic Assignments Using Aeromonas as a Test Case

    PubMed Central

    Colston, Sophie M.; Fullmer, Matthew S.; Beka, Lidia; Lamy, Brigitte

    2014-01-01

    ABSTRACT Prokaryotic taxonomy is the underpinning of microbiology, as it provides a framework for the proper identification and naming of organisms. The “gold standard” of bacterial species delineation is the overall genome similarity determined by DNA-DNA hybridization (DDH), a technically rigorous yet sometimes variable method that may produce inconsistent results. Improvements in next-generation sequencing have resulted in an upsurge of bacterial genome sequences and bioinformatic tools that compare genomic data, such as average nucleotide identity (ANI), correlation of tetranucleotide frequencies, and the genome-to-genome distance calculator, or in silico DDH (isDDH). Here, we evaluate ANI and isDDH in combination with phylogenetic studies using Aeromonas, a taxonomically challenging genus with many described species and several strains that were reassigned to different species as a test case. We generated improved, high-quality draft genome sequences for 33 Aeromonas strains and combined them with 23 publicly available genomes. ANI and isDDH distances were determined and compared to phylogenies from multilocus sequence analysis of housekeeping genes, ribosomal proteins, and expanded core genes. The expanded core phylogenetic analysis suggested relationships between distant Aeromonas clades that were inconsistent with studies using fewer genes. ANI values of ≥96% and isDDH values of ≥70% consistently grouped genomes originating from strains of the same species together. Our study confirmed known misidentifications, validated the recent revisions in the nomenclature, and revealed that a number of genomes deposited in GenBank are misnamed. In addition, two strains were identified that may represent novel Aeromonas species. PMID:25406383

  8. Complex Evolutionary History of the Aeromonas veronii Group Revealed by Host Interaction and DNA Sequence Data

    PubMed Central

    Faucher, Joshua; Horneman, Amy J.; Gogarten, J. Peter; Graf, Joerg

    2011-01-01

    Aeromonas veronii biovar sobria, Aeromonas veronii biovar veronii, and Aeromonas allosaccharophila are a closely related group of organisms, the Aeromonas veronii Group, that inhabit a wide range of host animals as a symbiont or pathogen. In this study, the ability of various strains to colonize the medicinal leech as a model for beneficial symbiosis and to kill wax worm larvae as a model for virulence was determined. Isolates cultured from the leech out-competed other strains in the leech model, while most strains were virulent in the wax worms. Three housekeeping genes, recA, dnaJ and gyrB, the gene encoding chitinase, chiA, and four loci associated with the type three secretion system, ascV, ascFG, aexT, and aexU were sequenced. The phylogenetic reconstruction failed to produce one consensus tree that was compatible with most of the individual genes. The Approximately Unbiased test and the Genetic Algorithm for Recombination Detection both provided further support for differing evolutionary histories among this group of genes. Two contrasting tests detected recombination within aexU, ascFG, ascV, dnaJ, and gyrB but not in aexT or chiA. Quartet decomposition analysis indicated a complex recent evolutionary history for these strains with a high frequency of horizontal gene transfer between several but not among all strains. In this study we demonstrate that at least for some strains, horizontal gene transfer occurs at a sufficient frequency to blur the signal from vertically inherited genes, despite strains being adapted to distinct niches. Simply increasing the number of genes included in the analysis is unlikely to overcome this challenge in organisms that occupy multiple niches and can exchange DNA between strains specialized to different niches. Instead, the detection of genes critical in the adaptation to specific niches may help to reveal the physiological specialization of these strains. PMID:21359176

  9. Complex evolutionary history of the Aeromonas veronii group revealed by host interaction and DNA sequence data.

    PubMed

    Silver, Adam C; Williams, David; Faucher, Joshua; Horneman, Amy J; Gogarten, J Peter; Graf, Joerg

    2011-02-16

    Aeromonas veronii biovar sobria, Aeromonas veronii biovar veronii, and Aeromonas allosaccharophila are a closely related group of organisms, the Aeromonas veronii Group, that inhabit a wide range of host animals as a symbiont or pathogen. In this study, the ability of various strains to colonize the medicinal leech as a model for beneficial symbiosis and to kill wax worm larvae as a model for virulence was determined. Isolates cultured from the leech out-competed other strains in the leech model, while most strains were virulent in the wax worms. Three housekeeping genes, recA, dnaJ and gyrB, the gene encoding chitinase, chiA, and four loci associated with the type three secretion system, ascV, ascFG, aexT, and aexU were sequenced. The phylogenetic reconstruction failed to produce one consensus tree that was compatible with most of the individual genes. The Approximately Unbiased test and the Genetic Algorithm for Recombination Detection both provided further support for differing evolutionary histories among this group of genes. Two contrasting tests detected recombination within aexU, ascFG, ascV, dnaJ, and gyrB but not in aexT or chiA. Quartet decomposition analysis indicated a complex recent evolutionary history for these strains with a high frequency of horizontal gene transfer between several but not among all strains. In this study we demonstrate that at least for some strains, horizontal gene transfer occurs at a sufficient frequency to blur the signal from vertically inherited genes, despite strains being adapted to distinct niches. Simply increasing the number of genes included in the analysis is unlikely to overcome this challenge in organisms that occupy multiple niches and can exchange DNA between strains specialized to different niches. Instead, the detection of genes critical in the adaptation to specific niches may help to reveal the physiological specialization of these strains.

  10. Precipitating antibody against Aeromonas salmonicida in serums of inbred albino Rainbow Trout (Salmo gairdneri)

    USGS Publications Warehouse

    Anderson, Douglas P.; Klontz, George W.

    1970-01-01

    Precipitins in albino rainbow trout serums were demonstrated by gel diffusion after a single parenteral exposure to the soluble antigens of Aeromonas salmonicida. The fraction of the serum containing antibody activity against the presented antigens was shown by immunoelectrophoresis to be in the nonmigrating region. This corresponded to the beta-2 fraction of rabbit serum. An antibody-containing component comparable with rabbit gamma globulin was not detected.

  11. Draft Genome Sequence of Aeromonas sobria Strain 08005, Isolated from Sick Rana catesbeiana

    PubMed Central

    Yang, Qiu-Hua; Zhou, Chen; Lin, Qi; Lu, Zhen; He, Li-Bin

    2017-01-01

    ABSTRACT Aeromonas sobria is a Gram-negative, rod-shaped, and ubiquitous bacterium. We present here the draft genome sequence of A. sobria strain 08005, isolated from an infected bullfrog. It is composed of 66 contigs totaling 4,678,951 bp, contains 4,252 coding DNA sequences (CDSs), four rRNAs, and 88 tRNA sequences, and shows the presence of various putative virulence-related genes. PMID:28082483

  12. Isolation of a pigment-producing strain of Aeromonas liquefaciens from silver salmon (Oncorhynchus kisutch)

    USGS Publications Warehouse

    Ross, A.J.

    1962-01-01

    Aeromonas salmonicida, the etiological agent of furunculosis in fish, is distinctive in the field of fish diseases in that it may readily be recognized by the water-soluble reddish-brown pigment formed on culture media containing tyrosine. Additional tests for the identification of this organism include blackening of the colonial growth when flooded with an aqueous solution of p-phenylenediamine and a lack of motility (Griffin, Progressive Fish Culturist 14:74, 1952).

  13. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    USGS Publications Warehouse

    Starliper, Clifford E.; Ketolab, Henry G.; Noyes, Andrew D.; Schill, William B.; Henson, Fred G.; Chalupnicki, Marc A.; Dittman, Dawn E.

    2015-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments for captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine if selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBC’s (0.02 to 0.04%) were obtained with three different sources of cinnamon oil. MBC’s for three sources of oregano and lemongrass oils ranged from 0.14 to 0.30% and 0.10 to 0.65%, respectively, and for two thyme oils were 2.11 and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBC’s to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBC’s for all but one isolate

  14. Prevalence and antimicrobial susceptibilities of Vibrio, salmonella, and Aeromonas isolates from various uncooked seafoods in Thailand.

    PubMed

    Woodring, Joseph; Srijan, Apichai; Puripunyakom, Paksathorn; Oransathid, Wilawan; Wongstitwilairoong, Boonchai; Mason, Carl

    2012-01-01

    Uncooked seafood samples were collected from open markets and supermarkets in Bangkok, Thailand, and were examined for the presence of Vibrio, Salmonella, and Aeromonas species from January to February 2008. From 120 samples, 272 bacterial isolates were identified through biochemical testing. Of all sea bass, shrimp, oyster, and blood cockle samples (30 of each) that were processed for culture, 114 (95%) samples had at least one detectable isolate of Vibrio, Salmonella, or Aeromonas, leaving only 6 (5%) samples free of them. All oyster sample (100%) had at least one pathogen, followed by sea bass (97%), blood cockles (97%), and shrimp (90%). Overall, 111 (92%) of all samples had detectable Vibrio spp., 32 (27%) had detectable Aeromonas spp., and 25 (21%) had detectable Salmonella enterica. There was no overall difference between positive samples collected from fresh markets versus supermarkets (relative risk, 0.97; 95% CI, 0.89 to 1.05). Resistance to ampicillin among isolated pathogens was relatively high (56%), while resistance to 12 other antibiotics, including azithromycin, ciprofloxacin, and trimethoprim-sulfamethoxazole, was relatively low (0, 0, and 3%, respectively). Study results indicate that uncooked seafood in Bangkok, Thailand, commonly harbors enteric pathogens and that consumption of uncooked seafood should be avoided to reduce foodborne illnesses.

  15. The Genus Aeromonas: Biochemical Characteristics, Atypical Reactions, and Phenotypic Identification Schemes

    PubMed Central

    Abbott, Sharon L.; Cheung, Wendy K. W.; Janda, J. Michael

    2003-01-01

    A total of 193 strains representing 14 different Aeromonas genomospecies were evaluated for 63 phenotypic properties to create useful tables for the reference identification of mesophilic aeromonads. Only 9 of 62 biochemical tests (14%) yielded uniform results, and the fermentation of certain carbohydrates was found to be linked to specific species. A number of unusual or aberrant properties for the genus Aeromonas were also detected in the collection of 428 strains (193 in the phenotypic study, 235 in a retrospective review). These tests included susceptibility to the vibriostatic agent, fermentation of m-inositol and d-xylose, hydrolysis of urea, and the lack of cytochrome oxidase activity. Fermentation of melibiose was linked to raffinose fermentation in all Aeromonas species except A. jandaei. Keys are provided for clinical laboratories choosing to identify aeromonads to species level based upon initial Møeller decarboxylase and dihydrolase reactions. In addition, several new tests were identified that help to separate members of the A. caviae complex (A. caviae, A. media, and A. eucreonophila). PMID:12791848

  16. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    USGS Publications Warehouse

    Starliper, Clifford E.; Ketola, H. George; Noyes, Andrew D.; Schill, William B.; Henson, Fred G.; Chalupnicki, Marc; Dittman, Dawn E.

    2015-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02–0.04%) were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate.

  17. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    PubMed

    Starliper, Clifford E; Ketola, Henry G; Noyes, Andrew D; Schill, William B; Henson, Fred G; Chalupnicki, Marc A; Dittman, Dawn E

    2015-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC's) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02-0.04%) were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate.

  18. A Disease Model of Muscle Necrosis Caused by Aeromonas dhakensis Infection in Caenorhabditis elegans

    PubMed Central

    Chen, Po-Lin; Chen, Yi-Wei; Ou, Chun-Chun; Lee, Tzer-Min; Wu, Chi-Jung; Ko, Wen-Chien; Chen, Chang-Shi

    2017-01-01

    A variety of bacterial infections cause muscle necrosis in humans. Caenorhabditis elegans has epidermis and bands of muscle that resemble soft-tissue structures in mammals and humans. Here, we developed a muscle necrosis model caused by Aeromonas dhakensis infection in C. elegans. Our data showed that A. dhakensis infected and killed C. elegans rapidly. Characteristic muscle damage in C. elegans induced by A. dhakensis was demonstrated in vivo. Relative expression levels of host necrosis-associated genes, asp-3, asp-4, and crt-1 increased significantly after A. dhakensis infection. The RNAi sensitive NL2099 rrf-3 (pk1426) worms with knockdown of necrosis genes of crt-1 and asp-4 by RNAi showed prolonged survival after A. dhakensis infection. Specifically knockdown of crt-1 and asp-4 by RNAi in WM118 worms, which restricted RNAi only to the muscle cells, conferred significant resistance to A. dhakensis infection. In contrast, the severity of muscle damage and toxicity produced by the A. dhakensis hemolysin-deletion mutant is attenuated. In another example, shiga-like toxin-producing enterohemorrhagic E. coli (EHEC) known to elicit toxicity to C. elegans with concomitant enteropathogenicty, did not cause muscle necrosis as A. dhakensis did. Taken together, these results show that Aeromonas infection induces muscle necrosis and rapid death of infected C. elegans, which are similar to muscle necrosis in humans, and then validate the value of the C. elegans model with A. dhakensis infection in studying Aeromonas pathogenicity. PMID:28101079

  19. Quorum Sensing Activity of Aeromonas Caviae Strain YL12, A Bacterium Isolated from Compost

    PubMed Central

    Lim, Yan-Lue; Ee, Robson; Yin, Wai-Fong; Chan, Kok-Gan

    2014-01-01

    Quorum sensing is a well-studied cell-to-cell communication method that involves a cell-density dependent regulation of genes expression mediated by signalling molecules. In this study, a bacterium isolated from a plant material compost pile was found to possess quorum sensing activity based on bioassay screening. Isolate YL12 was identified using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry and molecular typing using rpoD gene which identified the isolate as Aeromonas caviae. High resolution tandem mass spectrometry was subsequently employed to identify the N-acyl homoserine lactone profile of Aeromonas caviae YL12 and confirmed that this isolate produced two short chain N-acyl homoserine lactones, namely C4-HSL and C6, and the production was observed to be cell density-dependent. Using the thin layer chromatography (TLC) bioassay, both AHLs were found to activate C. violaceum CV026, whereas only C6-HSL was revealed to induce bioluminescence expression of E. coli [pSB401]. The data presented in this study will be the leading steps in understanding the role of quorum sensing in Aeromonas caviae strain YL12. PMID:24759107

  20. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    PubMed Central

    Starliper, Clifford E.; Ketola, Henry G.; Noyes, Andrew D.; Schill, William B.; Henson, Fred G.; Chalupnicki, Marc A.; Dittman, Dawn E.

    2014-01-01

    Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s) were determined for these four oils, Allimed® (garlic extract, Allium sativum) and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02–0.04%) were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5%) of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate. PMID:25685547

  1. Integron-containing IncU R plasmids pRAS1 and pAr-32 from the fish pathogen Aeromonas salmonicida.

    PubMed

    Sørum, Henning; L'Abée-Lund, Trine M; Solberg, Asta; Wold, Anette

    2003-04-01

    A 45-kb R plasmid, pRAS1, that confers resistance to tetracyclines, trimethoprim, and sulfonamides was isolated in 1989 from an atypical strain of the fish pathogen Aeromonas salmonicida. This plasmid could be transferred by conjugation to Escherichia coli with a high degree of efficiency (frequency, 0.48). The following year pRAS1 was isolated from A. salmonicida subsp. salmonicida in the same area. Incompatibility group U plasmid pRAS1 contained a drug resistance-determining region of 12 kb consisting of a class 1 integron similar to In4 of Tn1696 but with a dfrA16 gene cassette inserted. Close to IS6100 at the right end of Tn4 was a truncated Tn1721. Restriction enzyme analysis showed that R plasmid pAr-32, isolated from A. salmonicida in Japan in 1970, had the same backbone structure as pRAS1, while the drug resistance-determining region contained a complex class 1 integron with an aadA2 cassette; the chloramphenicol resistance gene catA2, as in In6 of pSa; and a duplicate of the 3' conserved segment of the integron.

  2. Aeromonas Diversity and Antimicrobial Susceptibility in Freshwater—An Attempt to Set Generic Epidemiological Cut-Off Values

    PubMed Central

    Baron, Sandrine; Granier, Sophie A.; Larvor, Emeline; Jouy, Eric; Cineux, Maelan; Wilhelm, Amandine; Gassilloud, Benoit; Le Bouquin, Sophie; Kempf, Isabelle; Chauvin, Claire

    2017-01-01

    The importance of the role of environment in the dissemination of antimicrobial resistant bacteria is now well recognized. Thus, bacterial indicators to monitor the phenomena are required. The Aeromonas genus is autochthonous in the aquatic environment and easy to detect in any water type, such as freshwater, or wastewater. These microorganisms are also causing infections in humans and animals (including fish). Furthermore, as Aeromonas spp. is able to acquire antimicrobial resistance mechanisms, it is candidate for indicator bacteria to follow antimicrobial resistance dissemination in aquatic environments. Unfortunately, to date, interpretation criteria for Aeromonas spp. for antimicrobial susceptibility tests are scarce in the literature. No epidemiological cut-off values for Aeromonas are currently available at EUCAST to interpret Minimum Inhibitory Concentrations (MIC). The only interpretation criteria available are clinical breakpoints from CLSI that are adapted from Enterobacteriaceae. Based on the results of MIC distributions obtained for a collection of environmental isolates of Aeromonas, this study aimed at proposing tentative epidemiological cut-off values (COWT) for Aeromonas spp. assessing whether the genus is an acceptable level of definition. Thus, 233 isolates collected from 16 rivers were identified at species level using Maldi-Tof (Bruker). Eleven different species were identified, the most abundant were A. bestiarum (n = 54), A. salmonicida (n = 45), A. sobria (n = 41), and A. eucrenophila (n = 37). 96-well micro-plates containing different concentrations of 15 antimicrobials, namely cefotaxime, ceftazidime, chloramphenicol, colistin, enrofloxacin, erythromycin, florfenicol, flumequine, gentamicin, nalidixic acid, oxolinic acid, streptomycin, temocillin, tetracycline, and trimethoprim-sulfamethoxazole, were prepared. The broth micro-dilution method was used to determine the antimicrobial susceptibility of each isolate. The estimation of COWT

  3. Population dynamics, antibiotics resistance and biofilm formation of Aeromonas and Vibrio species isolated from aquatic sources in Northern Malaysia.

    PubMed

    Odeyemi, Olumide A; Ahmad, Asmat

    2017-02-01

    This study aimed to compare population dynamics, antibiotic resistance and biofilm formation of Aeromonas and Vibrio species from seawater and sediment collected from Northern Malaysia. Isolates with different colony morphology were characterized using both biochemical and molecular methods before testing for antibiotic resistance and biofilm formation. Results obtained from this study showed that in Kedah, the population of Aeromonas isolated from sediment was highest in Pantai Merdeka (8.22 log CFU/ml), Pulau Bunting recorded the highest population of Aeromonas from sediment (8.43 log CFU/g). It was observed that Vibrio species isolated from seawater and sediment were highest in Kuala Sanglang (9.21 log CFU/ml). In Kuala Perlis, the population of Aeromonas isolated from seawater was highest in Jeti (7.94 log CFU/ml). Highest population of Aeromonas from sediment was recorded in Kampong Tanah Baru (7.99 log CFU/g). It was observed that Vibrio species isolated from seawater was highest in Padang Benta (8.42 log CFU/g) while Jeti Kuala Perlis had highest population of Vibrio isolated from sediment. It was observed that location does not influence population of Aeromonas. The results of the independent t - test revealed that there was no significant relationship between location and population of Vibrio (df = 10, t = 1.144, p > 0.05). The occurrence of biofilm formation and prevalence of antibiotic resistant Aeromonas and Vibrio species in seawater and sediment pose danger to human and aquatic animals' health.

  4. Development and application of real-time PCR assays for quantifying total and aerolysin gene-containing aeromonas in source, intermediate, and finished drinking water.

    PubMed

    Yu, Chang-Ping; Farrell, Sara K; Robinson, Bruce; Chu, Kung-Hui

    2008-02-15

    Aeromonas spp., opportunistic pathogens, are listed as a microbiological contaminant on the Environmental Protection Agency's (EPA) Drinking Water Contaminant Candidate List. Culture-based methods for identification and quantification of Aeromonas in drinking water are time-consuming and often fail to differentiate pathogenic species from nonpathogenic ones. This study reports successful development and applications of two real-time PCR assays, based on 16S rRNA gene sequences and a virulence gene (aerolysin gene), for rapid and effective quantification of total and aerolysin gene-containing Aeromonas spp. The assays successfully quantified total and aerolysin gene-containing Aeromonas in source, intermediate, and finished water samples collected from seven water works and one pilot plant. The effectiveness of Aeromonas removal by different drinking water treatment processes was examined by comparing the results obtained from the EPA culture-based method and developed real-time PCR assays. Regardless of the methods, our results indicated that conventional water treatment combination (prechlorination/ coagulation/sedimentation/rapid sand filtration) and membrane filtration alone could effectively remove Aeromonas. Slow sand filtration alone might not be effective. The removal efficiencies by different disinfection treatments were not determined, due to the lack of detectable Aeromonas. No Aeromonas was detected in samples with turbidity below 0.06 NTU.

  5. Characterization of genetic determinants involved in antibiotic resistance in Aeromonas spp. and fecal coliforms isolated from different aquatic environments.

    PubMed

    Carnelli, Alessandro; Mauri, Federica; Demarta, Antonella

    2017-03-02

    Aeromonas spp. and fecal coliforms, two abundant and cultivable bacterial populations that can be found in water ecosystems, might substantially contribute to the spread of antibiotic resistance. We investigated the presence and spread of transposons (elements that can move from one location to another in the genome), integrons (structures able to capture and incorporate gene cassettes) and resistance plasmids in strains isolated from polluted and unpolluted water. We recovered 231 Aeromonas and 250 fecal coliforms from water samplings with different degrees of pollution (hospital sewage, activated sludge of a wastewater treatment plant, river water before and after treatment and water from an alpine lake). Sixteen Aeromonas spp. and 22 fecal coliforms carried intI, coding for the site-specific integrase of class 1 integrons, while 22 Aeromonas spp. and 14 fecal coliforms carried tnpA, the transposase gene of the Tn3-family of replicative transposons. The majority of intI and tnpA-positive strains were phenotypically resistant to at least four antibiotics. Integrons and transposons were mainly located on mobilizable plasmids. Our results did not detect common mobile structures in the two populations and therefore relativize the role played by Aeromonas spp. as vectors of antimicrobial resistance determinants between water and commensal gut bacteria.

  6. Presence of Aeromonas spp in water from drinking-water- and wastewater-treatment plants in Mexico City.

    PubMed

    Villarruel-López, Angélica; Fernández-Rendón, Elizabeth; Mota-de-la-Garza, Lydia; Ortigoza-Ferado, Jorge

    2005-01-01

    The frequency of Aeromonas spp in three wastewater-treatment plants (WWTPs) and two drinking-water plants (DWPs) in México City was determined. Samples were taken throughout a year by the Moore's swab technique. A total of 144 samples were obtained from WWTPs and 96 from DWPs of both incoming and outflowing water. Aeromonas spp was isolated in 31% of the samples, from both kinds of sources. The technique used for the isolation of the pathogen was suitable for samples with high associate microbiota content and for those with a scarce microbial content. The presence of mesophilic-aerobic, coliform, and fecal-coliform organisms was investigated to determine whether there was any correlation with the presence of Aeromonas spp. Most samples from WWTP, which did not comply with the Mexican standards, had the pathogen, and some of the samples from the outflow of the DWP, which were within the limits set by the Mexican standards, also had Aeromonas spp. Most samples containing Aeromonas spp. had concentrations below 0.1 ppm residual chlorine, and the strains were resistant to 0.3 ppm, which supports the recommendation to increase the residual chlorine concentration to 0.5 to 1.0 ppm, as recommended by the Mexican standards.

  7. Temperate bacteriophage {phi}O18P from an Aeromonas media isolate: Characterization and complete genome sequence

    SciTech Connect

    Beilstein, Frauke

    2008-03-30

    A group of 74 Aeromonas isolates from surface water of three ponds in Bielefeld, Germany was screened for prophage induction after UV irradiation. The phage {phi}O18P was induced from the Aeromonas media isolate O18. {phi}O18P belongs to the Myoviridae phage family. The complete nucleotide sequence of the double stranded DNA genome of bacteriophage {phi}O18P consists of 33,985 bp. The genome has 5' protruding cohesive ends of 16 bases. On the {phi}O18P genome 46 open reading frames (orfs) were identified which are organized in the modules integration and regulation, replication, head, packaging, tail and lysis. Additionally the phage DNA includes a methylase gene. Comparison of the genome architecture with those of other bacteriophages revealed significant similarities to the P2 phage family and especially to the prophages of Aeromonas salmonicida and the Vibrio cholerae phage K139.

  8. Evidence of Increased Antibiotic Resistance in Phylogenetically-Diverse Aeromonas Isolates from Semi-Intensive Fish Ponds Treated with Antibiotics

    PubMed Central

    Patil, Hemant J.; Benet-Perelberg, Ayana; Naor, Alon; Smirnov, Margarita; Ofek, Tamir; Nasser, Ahmed; Minz, Dror; Cytryn, Eddie

    2016-01-01

    The genus Aeromonas is ubiquitous in aquatic environments encompassing a broad range of fish and human pathogens. Aeromonas strains are known for their enhanced capacity to acquire and exchange antibiotic resistance genes and therefore, are frequently targeted as indicator bacteria for monitoring antimicrobial resistance in aquatic environments. This study evaluated temporal trends in Aeromonas diversity and antibiotic resistance in two adjacent semi-intensive aquaculture facilities to ascertain the effects of antibiotic treatment on antimicrobial resistance. In the first facility, sulfadiazine-trimethoprim was added prophylactically to fingerling stocks and water column-associated Aeromonas were monitored periodically over an 11-month fish fattening cycle to assess temporal dynamics in taxonomy and antibiotic resistance. In the second facility, Aeromonas were isolated from fish skin ulcers sampled over a 3-year period and from pond water samples to assess associations between pathogenic strains to those in the water column. A total of 1200 Aeromonas isolates were initially screened for sulfadiazine resistance and further screened against five additional antimicrobials. In both facilities, strong correlations were observed between sulfadiazine resistance and trimethoprim and tetracycline resistances, whereas correlations between sulfadiazine resistance and ceftriaxone, gentamicin, and chloramphenicol resistances were low. Multidrug resistant strains as well as sul1, tetA, and intI1 gene-harboring strains were significantly higher in profiles sampled during the fish cycle than those isolated prior to stocking and these genes were extremely abundant in the pathogenic strains. Five phylogenetically distinct Aeromonas clusters were identified using partial rpoD gene sequence analysis. Interestingly, prior to fingerling stocking the diversity of water column strains was high, and representatives from all five clusters were identified, including an A. salmonicida cluster

  9. Historical record of Yersinia ruckeri and Aeromonas salmonicida among sea-run Atlantic salmon (Salmo salar) in the Penobscot River

    USGS Publications Warehouse

    Cipriano, R.C.; Coll, J.

    2005-01-01

    Despite restoration efforts, only about 2,000 Atlantic salmon (Salmo salar) salmon have annually returned to New England Rivers and more than 71% of these fish migrate to the Penobscot River alone. This report provides a historical compilation on the prevalence's of both Yersinia ruckeri, cause of enteric redmouth disease, and Aeromonas salmonicida, cause of furunculosis, among mature sea-run Atlantic salmon that returned to the Penobscot River from 1976 to 2003. Aeromonas salmonicida was detected in 28.6% and Yersinia ruckeri was detected among 50% of the yearly returns. Consequently, Atlantic salmon that return to the river are potential reservoirs of infection.

  10. Rapid biochemical screening for Salmonella, Shigella, Yersinia, and Aeromonas isolates from stool specimens.

    PubMed

    De Ryck, R; Struelens, M J; Serruys, E

    1994-06-01

    Four screens for the rapid (4 to 6 h) biochemical detection of pathogens from enteric isolation media are described. The Salmonella screen consisted of Kligler iron agar (KIA), motility-indole-urea-tryptophan-deamination semisolid medium (MIU-TDA), and the o-nitrophenyl-beta-D-galactopyranoside (ONPG) test; the Shigella screen consisted of KIA, MIU-TDA, the ONPG test, and the lysine decarboxylation-indole test; the Yersinia screen consisted of a rhamnose broth; the Aeromonas screen consisted of a xylose agar plate. When tested on 2,102 fresh isolates and 71 stock strains, the screens correctly detected 212 enteric pathogens (sensitivity, 100%), with a specificity of 98.1%.

  11. Effects of temperature on biochemical reactions and drug resistance of virulent and avirulent Aeromonas salmonicida

    USGS Publications Warehouse

    Hahnel, G.B.; Gould, R.W.

    1982-01-01

    Incubation temperatures of 11°, 18° and 28° did not substantially affect biochemical reactions of either virulent or avirulent forms of Aeromonas salmonicida subspecies salmonicida. The only change observed, amygdalin fermentation, was positive at 11° and 18° but negative at 28°C. Several isolates utilized sucrose, a characteristic not normally recognized for A. salmonicida subspecies salmonicida.Antimicrobial susceptibility screening indicated resistance to novobiocin increased at the higher incubation temperatures. Standardized drug sensitivity testing procedures and precise zone diameter interpretive standards for bacterial fish pathogens are needed.

  12. Virulence factors-pathogenicity relationships for Aeromonas species from clinical and food isolates.

    PubMed

    Pin, C; Morales, P; Marín, M L; Selgas, M D; García, M L; Casas, C

    1997-01-01

    The presence of virulence factors in 96 Aeromonas strains isolated from food and clinical samples was studied. Neither cytotoxic activity and hydrophobicity, not the presence of pili or an extra surface layer made it possible to establish differences between food and clinical strains. Statistical studies showed that cytotoxin production was associated with a positive Voges-Proskauer reaction, inability to ferment arabinose and a positive lysine decarboxylation. Therefore, when comparing cytotoxic clinical and food strains with lysine decarboxylation phenotype, there was a significant difference (p < 0.05) between the two groups. The association of a cytotoxin production and lysine decarboxylation character should thus be considered as a possible virulence marker.

  13. Comparative genomics profiling of clinical isolates of Aeromonas salmonicida using DNA microarrays

    PubMed Central

    Nash, John HE; Findlay, Wendy A; Luebbert, Christian C; Mykytczuk, Oksana L; Foote, Simon J; Taboada, Eduardo N; Carrillo, Catherine D; Boyd, Jessica M; Colquhoun, Duncan J; Reith, Michael E; Brown, Laura L

    2006-01-01

    Background Aeromonas salmonicida has been isolated from numerous fish species and shows wide variation in virulence and pathogenicity. As part of a larger research program to identify virulence genes and candidates for vaccine development, a DNA microarray was constructed using a subset of 2024 genes from the draft genome sequence of A. salmonicida subsp. salmonicida strain A449. The microarray included genes encoding known virulence-associated factors in A. salmonicida and homologs of virulence genes of other pathogens. We used microarray-based comparative genomic hybridizations (M-CGH) to compare selected A. salmonicida sub-species and other Aeromonas species from different hosts and geographic locations. Results Results showed variable carriage of virulence-associated genes and generally increased variation in gene content across sub-species and species boundaries. The greatest variation was observed among genes associated with plasmids and transposons. There was little correlation between geographic region and degree of variation for all isolates tested. Conclusion We have used the M-CGH technique to identify subsets of conserved genes from amongst this set of A. salmonicida virulence genes for further investigation as potential vaccine candidates. Unlike other bacterial characterization methods that use a small number of gene or DNA-based functions, M-CGH examines thousands of genes and/or whole genomes and thus is a more comprehensive analytical tool for veterinary or even human health research. PMID:16522207

  14. Polyphasic characterization of Aeromonas salmonicida isolates recovered from salmonid and non-salmonid fish

    USGS Publications Warehouse

    Diamanka, A.; Loch, T.P.; Cipriano, R.C.; Faisal, M.

    2013-01-01

    Michigan's fisheries rely primarily upon the hatchery propagation of salmonid fish for release in public waters. One limitation on the success of these efforts is the presence of bacterial pathogens, including Aeromonas salmonicida, the causative agent of furunculosis. This study was undertaken to determine the prevalence of A. salmonicida in Michigan fish, as well as to determine whether biochemical or gene sequence variability exists among Michigan isolates. A total of 2202 wild, feral and hatchery-propagated fish from Michigan were examined for the presence of A. salmonicida. The examined fish included Chinook salmon, Oncorhynchus tshawytscha (Walbaum), coho salmon, O. kisutcha (Walbaum), steelhead trout, O. mykiss (Walbaum), Atlantic salmon, Salmo salar L., brook trout, Salvelinus fontinalis (Mitchill), and yellow perch, Perca flavescens (Mitchill). Among these, 234 fish yielded a brown pigment-producing bacterium that was presumptively identified as A. salmonicida. Further phenotypic and phylogenetic analyses identified representative isolates as Aeromonas salmonicida subsp. salmonicida and revealed some genetic and biochemical variability. Logistic regression analyses showed that infection prevalence varied according to fish species/strain, year and gender, whereby Chinook salmon and females had the highest infection prevalence. Moreover, this pathogen was found in six fish species from eight sites, demonstrating its widespread nature within Michigan.

  15. Aeromonas punctata derived depolymerase that disrupts the integrity of Klebsiella pneumoniae capsule: optimization of depolymerase production.

    PubMed

    Bansal, Shruti; Soni, Sanjeev Kumar; Harjai, Kusum; Chhibber, Sanjay

    2014-07-01

    Formation of dense, highly hydrated biofilm structures pose a risk for public and environmental health. Extracellular polymeric substances encompassing biofilms offer 1000-fold greater resistance as compared to the planktonic cells. Using enzymes as anti-biofouling agents, will improve penetration of antimicrobials and increase susceptibility of biofilms to components of immune system. The challenge of using enzymes derived from unrelated bacteria for the degradation of capsular matrix of Klebsiella pneumoniae has not been dealt in the past. Thus, statistical optimization was done to enhance depolymerase production by Aeromonas punctata, directed against the exopolysaccharide matrix of Klebsiella pneumoniae B5055, capable of substituting the available phage borne depolymerase enzyme. Optimization via central composite design (CCD) resulted in 16-fold enhancement in depolymerase yield (166.65 µmoles ml(-1)  min(-1) ) over unoptimized medium. Out of the 19 variables, media composition giving maximum expression levels of the enzyme consisted of 1 mg ml(-1) galactose and ammonium chloride, 1.5 mg ml(-1) each of capsular polysaccharide (CPS) and magnesium sulfate. Tryptic peptide analysis of the purified 29 kDa band by Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) showed a high homology with a protein of unknown function from Aeromonas cavaie Ae398. Further improvements in the enzyme can lead to its successful development as prophylactic and/or a therapeutic agent.

  16. Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources.

    PubMed

    Odeyemi, Olumide A; Ahmad, Asmat

    2017-01-01

    This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics - Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole (25 μg), Nalixidic acid (30 μg), Trimethoprim (1.25 μg), Novobiocin (5 μg), Penicilin (10 μg) and Chloramphenicol (10 μg) was tested. The results obtained from this study reveal multi drug resistance pattern among the isolates. All the isolates were completely resistant to Ampicillin, Novobiocin, Sulphamethoxazole and Trimethoprim, respectively but susceptible to Tetracycline (100%), Kanamycin (5.7%), Gentamicin (5.7%) and Oxytetracycline (24.5%). Antibiotics phenotyping of the bacteria revealed 21 different phenotypes among the isolates.

  17. DISTRIBUTION OF SIX VIRULENCE FACTORS IN AEROMONAS SPECIES ISOLATED FROM US DRINKING WATER UTILITIES: A PCR IDENTIFICATION

    EPA Science Inventory

    Surveys of finished drinking water conducted by the U.S. EPA during 2000-2001, revealed 7 out of 16 water utilities encompassing four states, were contaminated with Aeromonas species. A Polymerase Chain reaction (PCR) based genetic characterization determined the presence of six...

  18. Molecular characterization of antibiotic resistance in Pseudomonas and Aeromonas isolates from catfish of the Mekong Delta, Vietnam.

    PubMed

    Nguyen, Hoang Nam Kha; Van, Thi Thu Hao; Nguyen, Huu Thinh; Smooker, Peter M; Shimeta, Jeff; Coloe, Peter J

    2014-07-16

    A collection of 116 motile Pseudomonas spp. and 92 Aeromonas spp. isolated from 15 Vietnamese intensive catfish farms was analyzed to examine the molecular antibiotic resistance characteristics and the transferability of resistance markers within and between species. High levels of resistance to ampicillin, trimethoprim/sulfamethoxazole, nalidixic acid, chloramphenicol, and nitrofurantoin were observed. The percentage of multiple drug resistance of Pseudomonas spp. and Aeromonas spp. isolates was 96.6% and 61.9%, respectively. The multiple antibiotic resistance (MAR) index mean values of 0.457 and 0.293 of Pseudomonas and Aeromonas isolates, respectively, indicated that these isolates were exposed to high risk sources of contamination where antibiotics were commonly used. Approximately 33% of Pseudomonas spp. and 28% of Aeromonas spp. isolates from catfish contained class 1 integrons, but no class 2 integrons were detected. Several common resistance genes including aadA, dfrA and catB were harbored in class 1 integrons. Large plasmids (>55 kb) were frequently detected in 50% and 71.4% of the plasmids extracted from Pseudomonas and Aeromonas isolates, respectively. Conjugation and transformation experiments demonstrated the successful transfer of all or part of the resistance phenotypes of catfish isolates to the recipient strains, including laboratory strains and strains isolated from this study. These results highlight the likely role of catfish bacteria as a reservoir of antibiotic resistant, Gram-negative bacteria harboring a pool of mobile genetic elements that can readily be transferred intra- and interspecies. To our knowledge, this is the first report on molecular characterization of antibiotic resistance of bacteria isolated from catfish in Vietnam.

  19. Butyrate production under aerobic growth conditions by engineered Escherichia coli.

    PubMed

    Kataoka, Naoya; Vangnai, Alisa S; Pongtharangkul, Thunyarat; Yakushi, Toshiharu; Matsushita, Kazunobu

    2017-01-11

    Butyrate is an important industrial platform chemical. Although several groups have reported butyrate production under oxygen-limited conditions by a native producer, Clostridium tyrobutylicum, and by a metabolically engineered Escherichia coli, efforts to produce butyrate under aerobic growth conditions have met limited success. Here, we constructed a novel butyrate synthetic pathway that functions under aerobic growth conditions in E. coli, by modifying the 1-butanol synthetic pathway reported previously. The pathway consists of phaA (acetyltransferase) and phaB (NADPH-dependent acetoacetyl-CoA reductase) from Ralstonia eutropha, phaJ ((R)-specific enoyl-CoA hydratase) from Aeromonas caviae, ter (trans-enoyl-CoA reductase) from Treponema denticola, and endogenous thioesterase(s) of E. coli. To evaluate the potential of this pathway for butyrate production, culture conditions, including pH, oxygen supply, and concentration of inorganic nitrogen sources, were optimized in a mini-jar fermentor. Under the optimal conditions, butyrate was produced at a concentration of up to 140 mM (12.3 g/L in terms of butyric acid) after 54 h of fed-batch culture.

  20. Cylindrospermopsin Biodegradation Abilities of Aeromonas sp. Isolated from Rusałka Lake.

    PubMed

    Dziga, Dariusz; Kokocinski, Mikolaj; Maksylewicz, Anna; Czaja-Prokop, Urszula; Barylski, Jakub

    2016-02-25

    The occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in freshwater reservoirs is a common phenomenon. However, the biodegradation of this toxin in environmental samples has been observed only occasionally. In this work the biodegradation ability of cylindrospermopsin was investigated based on isolates from lakes with previous cyanotoxin history. Bacterial strains were identified based on the 16S rDNA and rpoD gene comparison. CYN biodegradation was monitored using the HPLC method. The R6 strain identified as Aeromonas sp. was documented as being capable of CYN removal. This biodegradation was dependent on the pH and temperature. Additionally, the stimulation of the growth of the R6 strain in the presence of CYN was indicated. Our discovery supports the hypothesis that (in analogy to the well-known phenomenon of microcystin biodegradation) in lakes dominated by potential CYN-producing cyanobacteria, the processes of microbial utilization of this toxin may occur.

  1. Aeromonas proteolyrica bacteria in aerospace environments. [possible genetic alterations and effects on man

    NASA Technical Reports Server (NTRS)

    Foster, B. G.

    1974-01-01

    Preflight studies on Aeromonas proteolytica are reported to investigate the possibility of genetic alterations resulting in increased proteolysis in spacecraft environments. This organism may be present on human tissue and could pose medical problems if its endopeptidase and a hemolysin were to be produced in ususually high quantities or altered in such a way as to be more effective in their activities. Considered are: (1) Development of a nutrative holding medium for suspension of organisms; (2) the establishment of baseline information for the standardization of the assay for endopeptidase levels and hemolytic titers; (3) formulation of a method by which intracutaneous hemorrhage could be quantitated in guinea pig tissue; and (4) the responses of these organisms to parameters of spaceflight and experimentation.

  2. Aeromonas salmonicida subsp. salmonicida in the light of its type-three secretion system

    PubMed Central

    Vanden Bergh, Philippe; Frey, Joachim

    2014-01-01

    Aeromonas salmonicida subsp. salmonicida is an important pathogen in salmonid aquaculture and is responsible for the typical furunculosis. The type-three secretion system (T3SS) is a major virulence system. In this work, we review structure and function of this highly sophisticated nanosyringe in A. salmonicida. Based on the literature as well as personal experimental observations, we document the genetic (re)organization, expression regulation, anatomy, putative functional origin and roles in the infectious process of this T3SS. We propose a model of pathogenesis where A. salmonicida induces a temporary immunosuppression state in fish in order to acquire free access to host tissues. Finally, we highlight putative important therapeutic and vaccine strategies to prevent furunculosis of salmonid fish. PMID:24119189

  3. A Report of Peritonitis from Aeromonas sobria in a Peritoneal Dialysis (PD) Patient with Necrotizing Fasciitis.

    PubMed

    Janma, Jirayut; Linasmita, Patcharasarn; Changsirikulchai, Siribha

    2015-11-01

    A 70-years of age, male patient with underlying type 2 diabetes mellitus, hypertension, dyslipidemia and ischemic heart disease had undergone continuous ambulatory peritoneal dialysis (CAPD)for 3 years without any episodes of peritonitis. He was diagnosed with necrotizing fasciitis and later developed peritonitis after receiving a laceration from an aquatic injury suffered during the flood disaster of 2011. The blood culture, necrotic tissue and the clear dialysate collected upon admission had shown Aeromonas sobria. The route of peritonitis may be from the hematogenous spread of A. sobria resulting in necrotizing fasciitis. A. sobria should be considered as the pathogen of peritonitis in PD patients who have history of wounds from contaminated water. We suggest that the PD patients who present with septicemia and did not meet the criteria for peritonitis, the initial dialysate effluent should be sent for culture. The benefit of this is to allow early recognition and treatment of peritonitis.

  4. The Aeromonas caviae AHA0618 gene modulates cell length and influences swimming and swarming motility.

    PubMed

    Lowry, Rebecca C; Parker, Jennifer L; Kumbhar, Ramhari; Mesnage, Stephane; Shaw, Jonathan G; Stafford, Graham P

    2014-12-17

    Aeromonas caviae is motile via a polar flagellum in liquid culture, with a lateral flagella system used for swarming on solid surfaces. The polar flagellum also has a role in cellular adherence and biofilm formation. The two subunits of the polar flagellum, FlaA and FlaB, are posttranslationally modified by O-linked glycosylation with pseudaminic acid on 6-8 serine and threonine residues within the central region of these proteins. This modification is essential for the formation of the flagellum. Aeromonas caviae possesses the simplest set of genes required for bacterial glycosylation currently known, with the putative glycosyltransferase, Maf1, being described recently. Here, we investigated the role of the AHA0618 gene, which shares homology (37% at the amino acid level) with the central region of a putative deglycosylation enzyme (HP0518) from the human pathogen Helicobacter pylori, which also glycosylates its flagellin and is proposed to be part of a flagellin deglycosylation pathway. Phenotypic analysis of an AHA0618 A. caviae mutant revealed increased swimming and swarming motility compared to the wild-type strain but without any detectable effects on the glycosylation status of the polar flagellins when analyzed by western blot analysis or mass spectroscopy. Bioinformatic analysis of the protein AHA0618, demonstrated homology to a family of l,d-transpeptidases involved in cell wall biology and peptidoglycan cross-linking (YkuD-like). Scanning electron microscopy (SEM) and fluorescence microscopy analysis of the wild-type and AHA0618-mutant A. caviae strains revealed the mutant to be subtly but significantly shorter than wild-type cells; a phenomenon that could be recovered when either AHA0618 or H. pylori HP0518 were introduced. We can therefore conclude that AHA0618 does not affect A. caviae behavior by altering polar flagellin glycosylation levels but is likely to have a role in peptidoglycan processing at the bacterial cell wall, consequently altering

  5. Acinetobacter, Aeromonas, and Trichococcus populations dominate the microbial community within urban sewer infrastructure

    PubMed Central

    VandeWalle, J. L.; Goetz, G.W.; Huse, S.M.; Morrison, H. G.; Sogin, M.L.; Hoffmann, R.G.; Yan, K.; McLellan, S.L.

    2012-01-01

    We evaluated the population structure and temporal dynamics of the dominant community members within sewage influent from two wastewater treatment plants (WWTPs) in Milwaukee, WI. We generated >1.1M bacterial pyrotag sequences from the V6 hypervariable region of 16S rRNA genes from 38 influent samples and two samples taken upstream in the sanitary sewer system. Only a small fraction of pyrotags from influent samples (~15%) matched sequences from human fecal samples. The fecal components of the sewage samples included enriched pyrotag populations from Lactococcus and Enterobacteriaceae relative to their fractional representation in human fecal samples. In contrast to the large number of distinct pyrotags that represent fecal bacteria such as Lachnospiraceae and Bacteroides, only one or two unique V6 sequences represented Acinetobacter, Trichococcus and Aeromonas, which collectively account for nearly 35% of the total sewage community. Two dominant Acinetobacter V6 pyrotags (designated Acineto tag 1 and Acineto tag 2) fluctuated inversely with a seasonal pattern over a 3-year period, suggesting two distinct Acinetobacter populations respond differently to ecological forcings in the system. A single nucleotide change in the V6 pyrotags accounted for the difference in these populations and corresponded to two phylogenically distinct clades based on full-length sequences. Analysis of wavelet functions, derived from a mathematical model of temporal fluctuations, demonstrated that other abundant sewer associated populations including Trichococcus and Aeromonas had temporal patterns similar to either Acineto tag 1 or Acineto tag 2. Populations with related temporal fluctuations were found to significantly correlate with the same WWTP variables (5-day BOD, flow, ammonia, total phosphorous, and suspended solids). These findings illustrate that small differences in V6 sequences can represent phylogenetically and ecologically distinct taxa. This work provides insight into

  6. Infection of sea lamprey with an unusual strain of Aeromonas salmonicida

    USGS Publications Warehouse

    Diamanka, Arfang; Loch, Thomas P.; Cipriano, Rocco C.; Winters, Andrew D.; Faisal, Mohamed

    2014-01-01

    The invasion of the Laurentian Great Lakes by the fish-parasitic sea lamprey has led to catastrophic consequences, including the potential introduction of fish pathogens. Aeromonas salmonicida is a bacterial fish pathogen that causes devastating losses worldwide. Currently, there are five accepted subspecies of Aeromonas salmonicida: A. salmonicida subsp. salmonicida, masoucida, smithia, achromogenes, and pectinolytica. We discuss the discovery of an isolate of A. salmonicida that is pathogenic to rainbow trout (Oncorhynchus mykiss) and exhibits unique phenotypic and molecular characteristics. We examined 181 adult sea lamprey (Petromyzon marinus) from the Humber River (Lake Ontario watershed) and 162 adult sea lamprey from Duffins Creek (Lake Ontario watershed) during the spring seasons of 2005–11. Among those, 4/343 (1.2%) sea lamprey were culture positive for A. salmonicida, whereby biochemical and molecular studies identified three of the isolates as A. salmonicida subsp. salmonicida. The remaining isolate (As-SL1) recovered from Humber River sea lamprey was phenotypically more similar to A. salmonicida subsp. salmonicida than to the four other A. salmonicida subspecies. However, unlike A. salmonicida subsp. salmonicida, As-SL1 was sucrose positive, produced an acid-over-acid reaction on triple-sugar iron medium and did not amplify with A. salmonicida subsp. salmonicida specific primers. Phylogenetic analysis based on partial stretches of the 16S rRNA and DNA gyrase subunit B genes further confirmed that the As-SL1 isolate was not A. salmonicida subsp. masoucida, smithia, achromogenes, or pectinolytica. Based on our analyses, the As-SL1 isolate is either an unusual strain of A. salmonicida subsp. salmonicida or a novel A. salmonicida subspecies. The four A. salmonicida isolates that were recovered from sea lamprey were pathogenic to rainbow trout in experimental challenge studies. Our study also underscores the potential role of sea lamprey in the ecology of

  7. PATHOGENIC ESCHERICHIA COLI

    EPA Science Inventory

    Escherichia coli is a bacterial species which inhabits the gastrointestinal tract of man and warm-blooded animals. Because of the ubiquity of this bacterium in the intestinal flora, it serves as an important indicator organism of fecal contamination. E. coli, aside from serving a...

  8. Pathogenic Escherichia coli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Escherichia coli, a member of the Enterobacteriaceae family, is a part of the normal flora of the intestinal tract of humans and a variety of animals. E. coli strains are classified on the basis of antigenic differences in two surface components (serotyping), the somatic antigen (O) of the lipopoly...

  9. Emerging Escherichia Pathogen

    PubMed Central

    Permpalung, Nitipong; Sentochnik, Deborah E.

    2013-01-01

    Escherichia hermannii was first identified as a new species in 1982. It has rarely been reported as a human pathogen. We report the first case of E. hermannii as the sole pathogen in a catheter-related bloodstream infection. PMID:23740732

  10. RFLP-PCR analysis of the aroA gene as a taxonomic tool for the genus Aeromonas.

    PubMed

    Cascón Soriano, A; Anguita Castillo, J; Hernanz Moral, C; Sánchez Salazar, M; Yugueros Marcos, J; Naharro Carrasco, G

    1997-11-15

    The aroA gene has been identified as a target in screening for the presence of most Aeromonas species so far described by PCR. Synthetic oligonucleotide primers of 24 and 25 nucleotides were used by PCR assay to amplify a sequence of the aroA gene, which encodes 3-phosphoshikimate-1-carboxyvinyltransferase, a key enzyme of aromatic amino acids and folate biosynthetic pathway. A 1236-bp DNA fragment, representing most of the aroA gene, according to the nucleotide sequence of A. salmonicida, was amplified from all Aeromonas species tested, which represented most of the 14 hybridization groups. HaeII digestion of the 1236-bp fragment generated a restriction fragment length polymorphisms which could be used as a powerful tool for identification of aeromonads to the genus level.

  11. Differential partition of virulent Aeromonas salmonicida and attenuated derivatives possessing specific cell surface alterations in polymer aqueous-phase systems

    NASA Technical Reports Server (NTRS)

    Van Alstine, J. M.; Trust, T. J.; Brooks, D. E.

    1986-01-01

    Two-polymer aqueous-phase systems in which partitioning of biological matter between the phases occurs according to surface properties such as hydrophobicity, charge, and lipid composition are used to compare the surface properties of strains of the fish pathogen Aeromonas salmonicida. The differential ability of strains to produce a surface protein array crucial to their virulence, the A layer, and to produce smooth lipopolysaccharide is found to be important in the partitioning behavior of Aeromonas salmonicida. The presence of the A layer is shown to decrease the surface hydrophilicity of the pathogen, and to increase specifically its surface affinity for fatty acid esters of polyethylene glycol. The method has application to the analysis of surface properties crucial to bacterial virulence, and to the selection of strains and mutants with specific surface characteristics.

  12. Molecular Phylogenetics and Temporal Diversification in the Genus Aeromonas Based on the Sequences of Five Housekeeping Genes

    PubMed Central

    Lorén, J. Gaspar; Farfán, Maribel; Fusté, M. Carmen

    2014-01-01

    Several approaches have been developed to estimate both the relative and absolute rates of speciation and extinction within clades based on molecular phylogenetic reconstructions of evolutionary relationships, according to an underlying model of diversification. However, the macroevolutionary models established for eukaryotes have scarcely been used with prokaryotes. We have investigated the rate and pattern of cladogenesis in the genus Aeromonas (γ-Proteobacteria, Proteobacteria, Bacteria) using the sequences of five housekeeping genes and an uncorrelated relaxed-clock approach. To our knowledge, until now this analysis has never been applied to all the species described in a bacterial genus and thus opens up the possibility of establishing models of speciation from sequence data commonly used in phylogenetic studies of prokaryotes. Our results suggest that the genus Aeromonas began to diverge between 248 and 266 million years ago, exhibiting a constant divergence rate through the Phanerozoic, which could be described as a pure birth process. PMID:24586399

  13. Identification of antibiotic resistance cassettes in class 1 integrons in Aeromonas spp. strains isolated from fresh fish (Cyprinus carpio L.).

    PubMed

    Sarria-Guzmán, Yohanna; López-Ramírez, María Patricia; Chávez-Romero, Yosef; Ruiz-Romero, Erick; Dendooven, Luc; Bello-López, Juan Manuel

    2014-05-01

    Forty-six Aeromonas spp. strains were isolated from fresh fish and investigated for their antimicrobial susceptibility, detection of Class 1 integrons by PCR, and arrangement of gene cassettes. Selected isolates were further characterized by enterobacterial repetitive intergenic consensus-PCR. Twenty isolates were found to carry Class 1 integrons. Amplification of the variable regions of the integrons revealed diverse bands ranging in size from 150 to 1,958 pb. Sequence analysis of the variable regions revealed the presence of several gene cassettes, such as adenylyl transferases (aadA2 and aadA5), dihydrofolate reductases (dfrA17 and dfrA1), chloramphenicol acetyl transferase (catB3), β-lactamase (oxa2), lincosamide nucleotidil transferase (linF), aminoglycoside-modifying enzyme (apha15), and oxacillinase (bla OXA-10). Two open reading frames with an unknown function were identified as orfC and orfD. The aadA2 cassette was the most common integron found in this study. Interestingly, five integrons were detected in the plasmids that might be involved in the transfer of resistance genes to other bacteria. This is a first report of cassette encoding for lincosamides (linF) resistance in Aeromonas spp. Implications on the incidence of integrons in isolates of Aeromonas spp. from fresh fish for human consumption, and its possible consequences to human health are discussed.

  14. Use of Aeromonas spp. as General Indicators of Antimicrobial Susceptibility among Bacteria in Aquatic Environments in Thailand

    PubMed Central

    Usui, Masaru; Tagaki, Chie; Fukuda, Akira; Okubo, Torahiko; Boonla, Chanchai; Suzuki, Satoru; Seki, Kanako; Takada, Hideshige; Tamura, Yutaka

    2016-01-01

    Antimicrobials are widely used, not only for treating human infections, but also for treatment of livestock and in fish farms. Human habitats in Southeastern Asian countries are located in close proximity to aquatic environments. As such, the human populations within these regions are at risk of exposure to antimicrobial resistant bacteria, and thereby disseminating antimicrobial resistance genes (ARGs). In this study, we collected water samples from 15 sites (5 sites in Chao Phraya River, 2 sites at the mouth of Chao Phraya River, 3 sites in Ta Chin River, and 5 sites at city canals) and 12 sites (6 sites at city canals; 2 sites at chicken farms; 2 sites at pig farms; and 2 samples from sites at pig farms, which were subsequently treated at a biogas plant) in Thailand in 2013 and 2014, respectively. In total, 117 Aeromonas spp. were isolated from the water samples, and these organisms exhibited various antimicrobial susceptibility profiles. Notably, there was a significant correlation between the environmental concentration of tetracyclines and the rates of tetracycline resistance in the isolated Aeromonas spp.; however, both the concentration and rates of tetracycline resistance in samples derived from pig farms were higher than those of samples harvested from other aquatic environments. These findings suggest that the high concentrations of antimicrobials observed in these aquatic environments likely select for ARGs. Furthermore, they indicate that Aeromonas spp. comprise an effective marker for monitoring antimicrobial resistance in aquatic environments. PMID:27433156

  15. Use of Aeromonas spp. as General Indicators of Antimicrobial Susceptibility among Bacteria in Aquatic Environments in Thailand.

    PubMed

    Usui, Masaru; Tagaki, Chie; Fukuda, Akira; Okubo, Torahiko; Boonla, Chanchai; Suzuki, Satoru; Seki, Kanako; Takada, Hideshige; Tamura, Yutaka

    2016-01-01

    Antimicrobials are widely used, not only for treating human infections, but also for treatment of livestock and in fish farms. Human habitats in Southeastern Asian countries are located in close proximity to aquatic environments. As such, the human populations within these regions are at risk of exposure to antimicrobial resistant bacteria, and thereby disseminating antimicrobial resistance genes (ARGs). In this study, we collected water samples from 15 sites (5 sites in Chao Phraya River, 2 sites at the mouth of Chao Phraya River, 3 sites in Ta Chin River, and 5 sites at city canals) and 12 sites (6 sites at city canals; 2 sites at chicken farms; 2 sites at pig farms; and 2 samples from sites at pig farms, which were subsequently treated at a biogas plant) in Thailand in 2013 and 2014, respectively. In total, 117 Aeromonas spp. were isolated from the water samples, and these organisms exhibited various antimicrobial susceptibility profiles. Notably, there was a significant correlation between the environmental concentration of tetracyclines and the rates of tetracycline resistance in the isolated Aeromonas spp.; however, both the concentration and rates of tetracycline resistance in samples derived from pig farms were higher than those of samples harvested from other aquatic environments. These findings suggest that the high concentrations of antimicrobials observed in these aquatic environments likely select for ARGs. Furthermore, they indicate that Aeromonas spp. comprise an effective marker for monitoring antimicrobial resistance in aquatic environments.

  16. Normal intestinal flora of wild Nile crocodiles (Crocodylus niloticus) in the Okavango Delta, Botswana.

    PubMed

    Lovely, C J; Leslie, A J

    2008-06-01

    Bacterial and fungal cultures were performed from cloacal swabs collected from 29 wild Nile crocodiles, captured in the Okavango Delta, Botswana. Sixteen species of bacteria and 6 fungal species were cultured. Individual crocodiles yielded 1-4 bacterial species, and 0-2 fungal species. The most commonly isolated bacteria were Microbacterium, Enterococcus faecalis, Aeromonas hydrophila, and Escherichia coli. No salmonellae were cultured. The most commonly occurring fungus was Cladosporium. Several of the bacterial and fungal species isolated have been implicated in cases of septicaemia in crocodilians. Knowledge of the normal intestinal flora will contribute towards the development of a crocodile-specific probiotic for use in farmed crocodiles.

  17. Aerobic intestinal flora of wild-caught African dwarf crocodiles Osteolaemus tetraspis.

    PubMed

    Huchzermeyer, F W; Henton, M M; Riley, J; Agnagna, M

    2000-09-01

    Intestinal contents were collected from wild-caught African dwarf crocodiles (Osteolaemus tetraspis) in 1993 and 1995 which were slaughtered at urban markets in the Congo Republic. The samples were kept frozen and brought back to Onderstepoort for aerobic culture. Out of 29 specimens, 33 species of bacteria and 20 species of fungi were isolated. The bacteria included three isolates of Salmonella and eight isolates of Escherichia coli, most of the latter being rough strains. The flora of individual specimens contained 1-5 bacterial and 0-5 fungal species. Neither Aeromonas hydrophila nor Edwardsiella tarda were isolated from any of the samples.

  18. Influence of Air Quality on the Composition of Microbial Pathogens in Fresh Rainwater

    PubMed Central

    Kaushik, Rajni; de la Cruz, Armah A.

    2012-01-01

    In this study, the microbiological quality of fresh rainwater was assessed from 50 rain events under tropical weather conditions for a year. The levels of four major opportunistic waterborne pathogens, namely, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Aeromonas hydrophila, in rainwater samples were quantified by using a robust and sensitive quantitative PCR (qPCR) method. Of the 50 rainwater samples, 25 were found to be positive for at least one pathogen: 21 for E. coli, 16 for P. aeruginosa, 6 for K. pneumoniae, and 1 for A. hydrophila. In addition to the microbiological assessment of rainwater samples, we also studied the influence of prevailing air quality on the microbial quality of rainwater over the sampling period. A significant change in the diversity and relative abundance of the basic microbial indicator organisms in rainwater was observed during a major regional air pollution episode in Southeast Asia due to biomass-burning emissions. PMID:22327595

  19. Adhesion of Pathogenic Bacteria to Food Contact Surfaces: Influence of pH of Culture

    PubMed Central

    Mafu, Akier Assanta; Plumety, Corinne; Deschênes, Louise; Goulet, Jacques

    2011-01-01

    The adhesion of Aeromonas hydrophila, Escherichia coli O157:H7, Salmonella Enteritidis, and Staphylococcus aureus to hydrophobic and hydrophilic surfaces in cultures with different pHs (6, 7, and 8) was studied. The results indicated that the type of material had no effect on the attachment capacity of microorganisms, while environmental pH influenced the adhesion of A. hydrophila, E. coli, and S. aureus to both solid substrates. The attachment of S. Enteritidis (P > .05) was not affected by the type of substrate or the culture pH, whereas E. coli displayed the weakest affinity for both polystyrene and glass surfaces. No correlation was established between the physicochemical properties of the materials, or the bacterial and the rate of bacterial adhesion, except for S. aureus. Photomicrographs have shown that surfaces were contaminated by small clusters of S. Enteritidis while S. aureus invaded the food contact surfaces in the form of small chains or cell aggregates. PMID:20981289

  20. Microbial biosafety of pilot-scale bioreactor treating MTBE and TBA-contaminated drinking water supply.

    PubMed

    Schmidt, Radomir; Klemme, David A; Scow, Kate; Hristova, Krassimira

    2012-03-30

    A pilot-scale sand-based fluidized bed bioreactor (FBBR) was utilized to treat both methyl tert-butyl ether (MTBE) and tert-butyl alcohol (TBA) from a contaminated aquifer. To evaluate the potential for re-use of the treated water, we tested for a panel of water quality indicator microorganisms and potential waterborne pathogens including total coliforms, Escherichia coli, Salmonella and Shigella spp., Campylobacter jejuni, Aeromonas hydrophila, Legionella pneumophila, Vibrio cholerae, Yersinia enterocolytica and Mycobacterium avium in both influent and treated waters from the bioreactor. Total bacteria decreased during FBBR treatment. E. coli, Salmonella and Shigella spp., C. jejuni, V. cholerae, Y. enterocolytica and M. avium were not detected in aquifer water or bioreactor treated water samples. For those pathogens detected, including total coliforms, L. pneumophila and A. hydrophila, numbers were usually lower in treated water than influent samples, suggesting removal during treatment. The detection of particular bacterial species reflected their presence or absence in the influent waters.

  1. Escherichia coli biofilms

    PubMed Central

    Beloin, Christophe; Roux, Agnès; Ghigo, Jean-Marc

    2008-01-01

    Escherichia coli is a predominant species among facultative anaerobic bacteria of the gastrointestinal tract. Both its frequent community lifestyle and the availability of a wide array of genetic tools contributed to establish E. coli as a relevant model organism for the study of surface colonization. Several key factors, including different extracellular appendages, are implicated in E. coli surface colonization and their expression and activity are finely regulated, both in space and time, to ensure productive events leading to mature biofilm formation. This chapter will present known molecular mechanisms underlying biofilm development in both commensal and pathogenic E. coli. PMID:18453280

  2. Production of hexaoligochitin from colloidal chitin using a chitinase from Aeromonas schubertii.

    PubMed

    Liu, Chao-Lin; Lan, Chih-Yu; Fu, Chuan-Chieh; Juang, Ruey-Shin

    2014-08-01

    Chitin derivatives, such as those with modified main saccharide chains and deacetylated side chains, exhibit versatile biological functions. The biomedical properties of chitin oligosaccharides depend on their degree of oligomerization. Of the chitin oligosaccharides, chitin hexamers are generally the most potent. In our recent study, N-acetylchitohexaose was obtained by digesting chitin with ASCHI61, a chitinase from Aeromonas schubertii. In this work, the factors involved in the production of chitin hexasaccharide were evaluated experimentally. Using steep map analysis and cross-analysis, the substrate concentration and reaction pH were identified as the key factors in this reaction, and the interactions between these parameters were observed. Using a response surface experimental design, we predicted that a colloidal chitin concentration of 3.4mgmL(-1) and a pH of 6.54 were the optimal conditions for producing hexaoligochitin. These conditions were verified in separate experiments, in which 38.73mmolL(-1) of N-acetylchitohexaose was obtained. The maximum amount of hexamer produced was 42.175mgL(-1), an increase of only 0.27% from the predicted value.

  3. Identification and pathogenicity of Aeromonas sobria on tail-rot disease in juvenile tilapia Oreochromis niloticus.

    PubMed

    Li, Ya; Cai, Shuang-Hu

    2011-02-01

    Thirty-six strains, numbered from PY01 to PY36, were isolated from six moribund Oreochromis niloticus. The biochemical characteristics of all strains conformed to the species description of Aeromonas sobria on the basis of API 20E and Biolog GN system. Furthermore, gyrB sequence of strain PY36 was sequenced and showed high similarity (99.8%) with A. sobria in Genbank. Antibiotic-resistance of strain PY36 was assessed by the Kirby-Bauer disk diffusion method, and the results showed it was susceptible and moderately susceptible to 12 and 3 of the 19 antimicrobials tested. Virulence of strain PY36 to juvenile tilapia was also tested, and we found that LD₅₀ was about 4.17 × 10³ CFU per fish in intraperitoneal injection. This is the first article to report that A. sobria was the pathogenic agent of tail-rot disease in juvenile tilapia. A. sobria was multi-resistant to the most frequently used antimicrobial drugs in China, so the antimicrobial resistance test should be carried out when these bacteria are isolated from biological samples in order to avoid therapeutic failures and spread of the pathogenic organisms in the environment.

  4. Cloning, sequencing and expression of the gene encoding the extracellular metalloprotease of Aeromonas caviae.

    PubMed

    Kawakami, K; Toma, C; Honma, Y

    2000-01-01

    A gene (apk) encoding the extracellular protease of Aeromonas caviae Ae6 has been cloned and sequenced. For cloning the gene, the DNA genomic library was screened using skim milk LB agar. One clone harboring plasmid pKK3 was selected for sequencing. Nucleotide sequencing of the 3.5 kb region of pKK3 revealed a single open reading frame (ORF) of 1,785 bp encoding 595 amino acids. The deduced polypeptide contained a putative 16-amino acid signal peptide followed by a large propeptide. The N-terminal amino acid sequence of purified recombinant protein (APK) was consistent with the DNA sequence. This result suggested a mature protein of 412 amino acids with a molecular mass of 44 kDa. However, the molecular mass of purified recombinant APK revealed 34 kDa by SDS-PAGE, suggesting that further processing at the C-terminal region took place. The 2 motifs of zinc binding sites deduced are highly conserved in the APK as well as in other zinc metalloproteases including Vibrio proteolyticus neutral protease, Emp V from Vibrio vulnificus, HA/P from Vibrio cholerae, and Pseudomonas aeruginosa elastase. Proteolytic activity was inhibited by EDTA, Zincov, 1,10-phenanthroline and tetraethylenepentamine while unaffected by the other inhibitors tested. The protease showed maximum activity at pH 7.0 and was inactivated by heating at 80 C for 15 min. These results together suggest that APK belongs to the thermolysin family of metalloendopeptidases.

  5. Aeromonas salmonicida Ati2 is an effector protein of the type three secretion system.

    PubMed

    Dallaire-Dufresne, Stéphanie; Barbeau, Xavier; Sarty, Darren; Tanaka, Katherine H; Denoncourt, Alix M; Lagüe, Patrick; Reith, Michael E; Charette, Steve J

    2013-09-01

    The bacterium Aeromonas salmonicida, a fish pathogen, uses the type three secretion system (TTSS) to inject effector proteins into host cells to promote the infection. The study of the genome of A. salmonicida has revealed the existence of Ati2, a potential TTSS effector protein. In the present study, a structure-function analysis of Ati2 has been done to determine its role in the virulence of A. salmonicida. Biochemical assays revealed that Ati2 is secreted into the medium in a TTSS-dependent manner. Protein sequence analyses, molecular modelling and biochemical assays demonstrated that Ati2 is an inositol polyphosphate 5-phosphatase, which hydrolyses PtdIns(4,5)P2 and PtdIns(3,4,5)P3 in a way similar to VPA0450, a protein from Vibrio parahaemolyticus having high sequence similarity with Ati2. Mutants of Ati2 with altered amino acids at two different locations in the catalytic site displayed no phosphatase activity. Wild-type and mutant forms of Ati2 were cloned into expression systems for Dictyostelium discoideum, a soil amoeba used as an alternative host to study A. salmonicida virulence. Expression tests allowed us to demonstrate that Ati2 is toxic for the host cell in a catalytic-dependent manner. Finally, this study demonstrated the existence of a new TTSS effector protein in A. salmonicida.

  6. Detection and quantification of Aeromonas salmonicida in fish tissue by real-time PCR.

    PubMed

    Bartkova, S; Kokotovic, B; Skall, H F; Lorenzen, N; Dalsgaard, I

    2017-02-01

    Furunculosis, a septicaemic infection caused by the bacterium Aeromonas salmonicida subsp. salmonicida, currently causes problems in Danish seawater rainbow trout production. Detection has mainly been achieved by bacterial culture, but more rapid and sensitive methods are needed. A previously developed real-time PCR assay targeting the plasmid encoded aopP gene of A. salmonicida was, in parallel with culturing, used for the examination of five organs of 40 fish from Danish freshwater and seawater farms. Real-time PCR showed overall a higher frequency of positives than culturing (65% of positive fish by real-time PCR compared to 30% by a culture approach). Also, no real-time PCR-negative samples were found positive by culturing. A. salmonicida was detected by real-time PCR, though not by culturing, in freshwater fish showing no signs of furunculosis, indicating possible presence of carrier fish. In seawater fish examined after an outbreak and antibiotics treatment, real-time PCR showed the presence of the bacterium in all examined organs (1-482 genomic units mg(-1) ). With a limit of detection of 40 target copies (1-2 genomic units) per reaction, a high reproducibility and an excellent efficiency, the present real-time PCR assay provides a sensitive tool for the detection of A. salmonicida.

  7. A Tale of Transmission: Aeromonas veronii Activity within Leech-Exuded Mucus

    PubMed Central

    Ott, Brittany M.; Dacks, Andrew M.; Ryan, Kenneth J.

    2016-01-01

    Transmission, critical to the establishment and persistence of host-associated microbiotas, also exposes symbionts to new environmental conditions. With horizontal transmission, these different conditions represent major lifestyle shifts. Yet genome-wide analyses of how microbes adjust their transcriptomes toward these dramatic shifts remain understudied. Here, we provide a comprehensive and comparative analysis of the global transcriptional profiles of a symbiont as it shifts between lifestyles during transmission. The gammaproteobacterium Aeromonas veronii is transmitted from the gut of the medicinal leech to other hosts via host mucosal castings, yet A. veronii can also transition from mucosal habitancy to a free-living lifestyle. These three lifestyles are characterized by distinct physiological constraints and consequently lifestyle-specific changes in the expression of stress-response genes. Mucus-bound A. veronii had the greatest expression in terms of both the number of loci and levels of transcription of stress-response mechanisms. However, these bacteria are still capable of proliferating within the mucus, suggesting the availability of nutrients within this environment. We found that A. veronii alters transcription of loci in a synthetic pathway that obtains and incorporates N-acetylglucosamine (NAG; a major component of mucus) into the bacterial cell wall, enabling proliferation. Our results demonstrate that symbionts undergo dramatic local adaptation, demonstrated by widespread transcriptional changes, throughout the process of transmission that allows them to thrive while they encounter new environments which further shape their ecology and evolution. PMID:26896136

  8. Inhibition of Aeromonas sobria serine protease (ASP) by α2-macroglobulin.

    PubMed

    Murakami, Yoji; Wada, Yoshihiro; Kobayashi, Hidetomo; Irie, Atsushi; Hasegawa, Makoto; Yamanaka, Hiroyasu; Okamoto, Keinosuke; Eto, Masatoshi; Imamura, Takahisa

    2012-10-01

    ASP is a serine protease secreted by Aeromonas sobria. ASP cleaves various plasma proteins, which is associated with onset of sepsis complications, such as shock and blood coagulation disorder. To investigate a host defense mechanism against this virulence factor, we examined the plasma for ASP inhibitor(s). Human plasma inhibited ASP activity for azocasein, which was almost completely abolished by treating plasma with methylamine, which inactivates α2-macroglobulin (α2-MG). The ASP-inhibitor complex in ASP-added plasma was not detected by immunoblotting using anti-ASP antibody; however, using gel filtration of the plasma ASP activity for an oligopeptide, the ASP substrate was eluted in the void fraction (Mw>200 000), suggesting ASP trapping by α2-MG. Indeed, human α2-MG inhibited ASP azocaseinolytic activity in a dose-dependent manner, rapidly forming a complex with the ASP. Fibrinogen degradation by ASP was completely inhibited in the presence of α2-MG. α1-Protease inhibitor, antithrombin, and α2-plasmin inhibitor neither inhibited ASP activity nor formed a complex with ASP. Surprisingly, ASP degraded these plasma serine protease inhibitors. Thus, α2-MG is the major ASP inhibitor in the human plasma and can limit ASP virulence activities in A. sobria infection sites. However, as shown by fluorescence correlation spectroscopy, slow ASP inhibition by α2-MG in plasma may indicate insufficient ASP control in vivo.

  9. Effect of a phytogenic feed additive on the susceptibility of Onchorhynchus mykiss to Aeromonas salmonicida.

    PubMed

    Menanteau-Ledouble, S; Krauss, I; Santos, G; Fibi, S; Weber, B; El-Matbouli, M

    2015-06-29

    In recent years, feed additives have increasingly been adopted by the aquaculture industry. These supplements not only offer an alternative to antibiotics but have also been linked to enhanced growth performance. However, the literature is still limited and provides contradictory information on their effectiveness. This is mainly due to the wide variety of available products and their complex mechanisms of action. Phytogenic feed additives have been shown to have antimicrobial effects and can improve growth performance. In the present study, we investigated the susceptibility of several fish pathogenic bacteria to a phytogenic essential oil product in vitro. In addition, we determined the protective effect of a commercial phytogenic feed additive containing oregano, anis and citrus oils on the resistance of rainbow trout Oncorhynchus mykiss to infection by Aeromonas salmonicida. The bacterium was administered through 3 different routes: intra-peritoneal injection, immersion in a bacterial solution and cohabitation with infected fish. Mortality rates were significantly lower in infected rainbow trout that had received the feed additive: the overall mortality rate across all routes of infection was 18% in fish fed a diet containing the additive compared to 37% in fish that received unsupplemented feed. The route of infection also significantly impacted mortality, with average mortality rates of 60, 17.5 and 5% for intra-peritoneal injection, immersion and cohabitation, respectively. In general, fish were better protected against infection by immersion than infection by injection.

  10. Carbapenemase-Producing Aeromonas veronii Disseminated in the Environment of an Equine Specialty Hospital.

    PubMed

    Adams, Rachael J; Mathys, Dimitria A; Mollenkopf, Dixie F; Whittle, Andrea; Daniels, Joshua B; Wittum, Thomas E

    2017-03-27

    The emergence of bacterial resistance to carbapenem antibiotics is an urgent public health threat. Carbapenem drugs are a last resort treatment option for life-threatening infections. The frequent use of broad-spectrum antibiotics to treat hospitalized patients provides significant selection pressure favoring the emergence and dissemination of resistant organisms, including carbapenem-resistant Enterobacteriaceae (CRE). CREs have been reported in animal populations, but only rarely in horses. Our objective was to determine the prevalence of CRE in the environment of a referral equine specialty hospital. Environmental samples were collected on seven different sampling dates. Four clonal carbapenemase-producing Aeromonas veronii were recovered from 315 sampled surfaces (1.3%). All four isolates harbored the carbapenemase-producing, metallo-β-lactamase gene blacphA, although corresponding minimum inhibitory concentrations were within the susceptible range for imipenem and meropenem. All had an identical multilocus sequence type with a previously unreported allelic profile and contained multiple plasmids. To our knowledge, this recovery of blacphA-harboring A. veronii is the first report of carbapenemase-producing bacteria in the environment of an equine veterinary hospital. However, the low recovery rate suggests that environmental contamination is uncommon. Appropriate hospital cleaning and disinfection protocols are necessary to maintain a low risk of contamination for patients and personnel.

  11. Growth inhibition of Aeromonas salmonicida and Yersinia ruckeri by disinfectants containing peracetic acid.

    PubMed

    Meinelt, Thomas; Phan, Thy-My; Behrens, Sascha; Wienke, Andreas; Pedersen, Lars-Flemming; Liu, Dibo; Straus, David L

    2015-04-08

    Peracetic acid (PAA) is a therapeutic agent used for disinfection in aquaculture, but it must be investigated thoroughly in order to mitigate diseases without harming the fish. Successful disinfectants (like PAA) should not leave dangerous residues in the environment in order to successfully contribute to sustainable aquaculture. The aim of our study was to compare the effectiveness of 6 commercial PAA products with different molecular PAA:H2O2 ratios to reduce bacterial growth of Aeromonas salmonicida and Yersinia ruckeri and to determine effective concentrations and exposure times. All products reduced colony-forming units (CFUs) of A. salmonicida and Y. ruckeri. Products with higher molecular PAA:H2O2 ratios inhibited growth better than products with lower molecular PAA:H2O2 ratios at the same PAA concentration; this indicates that H2O2 is not the driving force in the reduction of A. salmonicida and Y. ruckeri growth by PAA in vitro. The practical application of the products with high molecular PAA:H2O2 ratios should be prioritized if these pathogens are diagnosed.

  12. Diverse Profiles of N-acyl Homoserine L-Lactones, Biofilm, Virulence Genes and Integrons in Food-Borne Aeromonas Isolates.

    PubMed

    Nagar, Vandan; Sinha, Vibha; Bandekar, Jayant R

    2015-08-01

    Aeromonas are regarded as opportunistic as well as primary pathogens of humans and fish, and are associated with gastroenteritis and septicemia in humans. Production of N-acyl-homoserine lactone (AHL) signal molecules and biofilm was determined in 22 Aeromonas isolates, from different food products in India, using thin-layer chromatography (TLC) analysis and microtiter-plate assay, respectively. Overall, highly heterogeneous patterns of AHL production were observed, with the production of N-butanoyl homoserine lactone (C4-HSL) and N-hexanoyl homoserine lactone (C6-HSL) by the majority (81.8%) of Aeromonas food isolates. Moreover, putative N-pentanoyl homoserine lactone (C5-HSL), N-heptanoyl homoserine lactone (C7-HSL), and N-octanoyl homoserine lactone (C8-HSL) were produced by 72.7%, 27.3%, and 9.1% of isolates, respectively. This is the 1st report of production of C7-HSL by Aeromonas species. Aeromonas food isolates were highly variable in their biofilm forming abilities with majority of them as weak biofilm producers in 2 different media, TSB and M9 minimal medium supplemented with 0.4% glucose. The genes encoding for putative virulence factors, glycerophospholipid cholesterol acyltransferase (gcat), heat-labile cytotonic enterotoxin (alt), heat-stable cytotonic enterotoxin (ast), serine protease (ser), polar flagella (fla), and lateral flagella (lafA) were present in 95.5%, 59.1%, 22.7%, 81.8%, 77.3%, and 22.7% of the strains, respectively. Class 1 integrons (100 to 3000 bp) were found in 68.2% of food isolates; whereas, 50% isolates contained class 2 integrons (150 to 1600 bp). This study provides a baseline data on the diversity of AHLs, biofilm forming ability and presence of virulence genes and integrons in Aeromonas food isolates from India.

  13. Effects of bacteria on the growth of an amoeba infecting the gills of turbot.

    PubMed

    Paniagua, E; Paramá, A; Iglesias, R; Sanmartín, M L; Leiro, J

    2001-05-04

    We analysed the influence of various bacteria on the in vitro growth of trophozoites of a Platyamoeba strain isolated from diseased gill tissues of cultured turbot. Little or no growth was shown by amoebae cultured in the presence of (1) the turbot-pathogenic bacteria Vibrio anguillarum, Aeromonas salmonicida or Streptococcus sp., (2) Pasteurella piscicida or Vibrio vulnificus (pathogenic for some fishes but not turbot), or (3) the non-pathogenic 'environmental' bacteria Vibrio campbelli, Vibrio fluvialis or Pseudomonas dondorofii. The only bacteria which were successfully utilized as food sources were Aeromonas hydrophila (pathogenic for some fishes but not turbot) and the non-pathogens Vibrio natriegens, Pseudomonas nautica and Escherichia coli. These results suggest that the colonization of the gills of cultured turbot by the epizoic amoeba Platyamoeba may be an indicator of faecal contamination.

  14. Recent advances in the study of the taxonomy, pathogenicity, and infectious syndromes associated with the genus Aeromonas.

    PubMed Central

    Janda, J M

    1991-01-01

    Over the past decade, the emergence of Aeromonas species as bona fide human pathogens and their probable role as etiologic agents of bacterial gastroenteritis have resulted in an explosion of scientific interest in the genus. Major accomplishments occurring in this field during that interval include a more refined taxonomy, identification of new cell-associated factors (surface layers, pili), and the molecular analysis of selected extracellular gene products that may play a critical role in pathogenesis (hemolysins, enterotoxins). This review provides an updated overview of recent systematic, clinical, and pathophysiologic advances and defines key areas of medical and scientific interest in which major questions remain unanswered. PMID:1747858

  15. Aeromonas jandaei (formerly genospecies DNA group 9 A. sobria), a new sucrose-negative species isolated from clinical specimens.

    PubMed Central

    Carnahan, A; Fanning, G R; Joseph, S W

    1991-01-01

    A large numerical taxonomy study conducted in 1988 of 165 mostly clinical Aeromonas strains from diverse geographic sources produced a cluster (S = 84%, SSM) of four sucrose-negative strains that included the DNA definition strain for DNA group 9 A. sobria (CDC 0787-80). These four strains, together with five additional strains received in 1989, were subjected to DNA-DNA hybridization (hydroxyapatite, 32P, 60 and 75 degrees C), and all eight strains were closely related to the ninth labeled DNA group 9 definition strain CDC 0787-80 (73 to 86% relatedness at 60 degrees C and 68 to 80% relatedness at 75 degrees C; percent divergence, 2.0 to 3.5). Type strains and DNA definition strains for all other established Aeromonas species were only 35 to 72% related (60 degrees C) to CDC 0787-80. We propose the name Aeromonas jandaei for this highly related group of nine strains, formerly known as DNA group 9 A. sobria. The type strain was designated ATCC 49568 (CDC 0787-80). The nine strains were examined at 36 degrees C and were found to be resistant to 0/129 (vibriostatic agent) and uniformly positive for oxidase, gas production from glucose, indole, lysine decarboxylase, arginine dihydrolase, o-nitrophenyl-beta-D-galactopyranoside, motility (25 degrees C), nitrate reduction, citrate utilization, hemolysis on sheep blood agar, and growth in Trypticase soy broth with no added NaCl. They all fermented D-glucose, D-mannitol, and mannose but did not ferment sucrose, cellobiose, L-arabinose, inositol, salicin, or D-sorbitol. They were uniformly negative for esculin and urea hydrolysis, elastase production, ornithine decarboxylation, and the string test. The antibiogram of A. jandaei resembled that of other aeromonads (resistance to ampicillin and cephalothin), but it differed from most other aeromonads because of resistance to single dilution of colistin and differed from clinical A. veronii biogroup sorbria (formerly A. sobria) by its nearly uniform resistance to cephalothin

  16. Diarrheagenic Escherichia coli

    PubMed Central

    Nataro, James P.; Kaper, James B.

    1998-01-01

    Escherichia coli is the predominant nonpathogenic facultative flora of the human intestine. Some E. coli strains, however, have developed the ability to cause disease of the gastrointestinal, urinary, or central nervous system in even the most robust human hosts. Diarrheagenic strains of E. coli can be divided into at least six different categories with corresponding distinct pathogenic schemes. Taken together, these organisms probably represent the most common cause of pediatric diarrhea worldwide. Several distinct clinical syndromes accompany infection with diarrheagenic E. coli categories, including traveler’s diarrhea (enterotoxigenic E. coli), hemorrhagic colitis and hemolytic-uremic syndrome (enterohemorrhagic E. coli), persistent diarrhea (enteroaggregative E. coli), and watery diarrhea of infants (enteropathogenic E. coli). This review discusses the current level of understanding of the pathogenesis of the diarrheagenic E. coli strains and describes how their pathogenic schemes underlie the clinical manifestations, diagnostic approach, and epidemiologic investigation of these important pathogens. PMID:9457432

  17. Diarrheagenic Escherichia coli.

    PubMed

    Gomes, Tânia A T; Elias, Waldir P; Scaletsky, Isabel C A; Guth, Beatriz E C; Rodrigues, Juliana F; Piazza, Roxane M F; Ferreira, Luís C S; Martinez, Marina B

    2016-12-01

    Most Escherichia coli strains live harmlessly in the intestines and rarely cause disease in healthy individuals. Nonetheless, a number of pathogenic strains can cause diarrhea or extraintestinal diseases both in healthy and immunocompromised individuals. Diarrheal illnesses are a severe public health problem and a major cause of morbidity and mortality in infants and young children, especially in developing countries. E. coli strains that cause diarrhea have evolved by acquiring, through horizontal gene transfer, a particular set of characteristics that have successfully persisted in the host. According to the group of virulence determinants acquired, specific combinations were formed determining the currently known E. coli pathotypes, which are collectively known as diarrheagenic E. coli. In this review, we have gathered information on current definitions, serotypes, lineages, virulence mechanisms, epidemiology, and diagnosis of the major diarrheagenic E. coli pathotypes.

  18. Interaction of the fish pathogen Aeromonas salmonicida with rainbow trout macrophages.

    PubMed Central

    Garduño, R A; Kay, W W

    1992-01-01

    A procedure was developed to culture rainbow trout macrophages (M phi) on supported glass coverslips. Using this method and a variety of well-characterized Aeromonas salmonicida strains with normal or altered cell surfaces, we investigated the role of this unusual bacterial surface in the bacterium-M phi interaction. An intact crystalline protein array, the A-layer, mediated adherence of A. salmonicida cells to M phi even in the absence of opsonins. In contrast, unopsonized cells of an A-layer-negative (A-) mutant with a smooth lipopolysaccharide (LPS) layer were unable to interact with M phi. However, this ability was recovered when the A-layer was reconstituted onto the smooth LPS surface of these A- LPS+ cells. Two A. salmonicida mutants possessing the A-layer in different disorganized states had a reduced ability to interact with M phi. A+ cells grown under calcium limitation produced A-layers locked into an alternative conformation which mediated the highest levels of M phi association in the absence of opsonins or any other surface coating. Coating A+ cells with hemin greatly increased their levels of M phi association, and bacterial cells grown on trout blood agar plates also had a dramatic increase in their ability to interact with M phi. Only A+ A. salmonicida cells were highly cytotoxic to trout M phi, especially after being coated with hemin, presumably due to a more focused targeting of the bacterial cell onto the M phi surface and/or into the intracellular regions of the M phi. Images PMID:1398975

  19. Inhibition of biosynthesis of metalloprotease of Aeromonas sobria by sodium chloride in the medium.

    PubMed

    Takahashi, Eizo; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Nair, Gopinath Balakrish; Takeda, Yoshifumi; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

    2011-01-01

    The present authors have previously shown that the serine protease activity of Aeromonas sobria is markedly decreased when A. sobria is cultured in medium containing 3.0% sodium chloride (NaCl, concentration almost equivalent to sea water salinity), and that this occurs because, although the synthesis of ASP is not disturbed by the salt in the medium, the maturation pathway of serine protease of A. sobria (ASP) does not proceed successfully in such a medium. In this study, the effect of salt in the medium on the production of metalloprotease by A. sobria (AMP) was examined. A. sobria produced AMP in the milieu when the bacteria were cultured in medium containing (NaCl) at a concentration of 0.5%. However, AMP was not produced when the bacteria were cultured in salty medium containing 1.5% or more NaCl. To examine how NaCl reduces the production of metalloprotease by A. sobria, the amount of amp mRNA in the cell was measured and it was found that this decreased in proportion to the concentration of NaCl in the medium. The mRNA of amp was not detected in cells cultured in medium containing 1.5% or more NaCl. This means that the transcription of amp is inhibited in salty condition. As described, NaCl in the medium disturbs the maturation pathway of ASP. The mode of action whereby NaCl suppresses AMP activity in A. sobria differs from the mechanism for suppressing ASP activity.

  20. Synergistic iron reduction and citrate dissimilation by Shewanella alga and Aeromonas veronii

    PubMed

    Knight; Caccavo; Wudyka; Blakemore

    1996-10-17

    Two bacterial isolates from Great Bay Estuary, New Hampshire, in co-culture carried out anaerobic dissimilation of citric acid with Fe(III) as the terminal electron acceptor. Neither isolate oxidized citrate with Fe(III) anaerobically in axenic culture. The Fe(III) reducer, Shewanella alga strain BrY, did not grow anaerobically with citrate as an energy source. The citrate utilizer, Aeromonas veronii, did not reduce iron axenically with a variety of electron donors including citrate. The onset of iron reduction by the co-culture occurred after initiation of citrate dissimilation and just prior to initiation of growth by either organism (as measured by viable plate counts). Anaerobic culture growth rates and final cell densities of each bacterial strain were greater in co-culture than in axenic cultures. By 48 h of growth, the co-culture had consumed 27 mM citrate as compared with 12 mM dissimilated by the axenic culture of A. veronii. By 48 h the co-culture produced half as much formate (6 mM) and twice as much acetate (40 mM) as did A. veronii grown axenically (12 mM and 20 mM, respectively). Formate produced from citrate by A. veronii appeared to have supported growth and Fe(III) reduction by S. alga.Although not obligatory, nutrient coupling between these two organisms illustrates that fermentative (A. veronii-type) organisms can convert organic compounds such as citrate to those used as substrates by dissimilatory Fe(III) reducers, including S. alga. This synergism broadens the range of substrates available for iron reduction, stimulates the extent and rate of organic electron donor degradation (and that of iron reduction) and enhances the growth of each participant.

  1. The impact of Aeromonas salmonicida infection on innate immune parameters of Atlantic salmon (Salmo salar L).

    PubMed

    Du, Yishuai; Yi, Mengmeng; Xiao, Peng; Meng, Lingjie; Li, Xian; Sun, Guoxiang; Liu, Ying

    2015-05-01

    Enzyme activities and gene expression of a number of innate immune parameters in the serum, mucus and skin of Atlantic salmon (Salmo salar) were investigated after challenge with a pathogenic strain of Aeromonas salmonicida (A. salmonicida). Fish were injected in the dorsal muscle with either 100 μl bacterium solution, about 3.05 × 10(7) CFU/ml A. salmonicida, or 100 μl 0.9% NaCl (as control group) and tissue samples were collected at days 0, 2, 4 and 6 post-injection. Lysozyme (LSZ) and alkaline phosphatase (AKP) activities in serum, mucus and skin, and LSZ and AKP mRNA expression in skin of the challenged fish were higher than those of the control at most of the experimental time, with significant differences at several time points (P < 0.05), indicating the involvement of LSZ and AKP in the innate immunity of Atlantic salmon to A. salmonicida. Superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities in mucus and skin, along with the SOD, POD and CAT mRNA expression in skin significantly decreased at day 4 and 6, indicating the decreased antioxidant capacity of the challenged fish. Glutamate pyruvate transaminase (GPT) and glutamic oxalacetic transaminase (GOT) activities in serum, mucus and skin of the challenged group were all higher than those of the control after the injection, and at several time points significant differences were found between the two groups, suggesting organs of fish were impaired after the pathogen infection. The changes of the GPT and GOT activities could be used as potential biomarkers for the impairment of physiological functions caused by the pathogen infection. Identified biomarkers of the immune responses will contribute to the early-warning system of the disease. So this study will not only provide a theoretical basis for vaccine development, but also provide basic data for the establishment of early warning systems for diseases caused by A. salmonicida in Atlantic salmon rearing.

  2. Evaluation of commercially prepared transport systems for nonlethal detection of Aeromonas salmonicida in salmonid fish

    USGS Publications Warehouse

    Cipriano, R.C.; Bullock, G.L.

    2001-01-01

    In vitro studies indicated that commercially prepared transport systems containing Amies, Stuart's, and Cary-Blair media worked equally well in sustaining the viability of the fish pathogen Aeromonas salmonicida, which causes furunculosis. The bacterium remained viable without significant increase or decrease in cell numbers for as long as 48 h of incubation at 18-20??C in Stuart's transport medium; consequently, obtaining mucus samples in such tubes were comparable to on-site detection of A. salmonicida by dilution plate counts on Coomassie Brilliant Blue agar. In three different assays of 100 samples of mucus from Atlantic salmon Salmo salar infected subclinically with A. salmonicida, dilution counts conducted on-site proved more reliable for detecting the pathogen than obtaining the samples in the transport system. In the on-site assays, dilution counts detected the pathogen in 34, 41, and 22 samples, whereas this was accomplished in only 15, 15, and 3 of the respective samples when the transport system was used. In an additional experiment, Arctic char Salvelinus alpinus sustaining a frank epizootic of furunculosis were sampled similarly. Here, too, dilution counts were more predictive of the prevalence of A. salmonicida and detected the pathogen in 46 mucus samples; in comparison, only 6 samples collected by using the transport system were positive. We also observed that the transport system supported the growth of the normal mucus bacterial flora. Particularly predominant among these were motile aeromonads and Pseudomonas fluorescens. In studies of mixed culture growth, two representatives of both of the latter genera of bacteria outgrew A. salmonicida - in some cases, to the total exclusion of the pathogen itself.

  3. The catalytic role of glutamate 151 in the leucine aminopeptidase from Aeromonas proteolytica.

    PubMed

    Bzymek, Krzysztof P; Holz, Richard C

    2004-07-23

    Glutamate 151 has been proposed to act as the general acid/base during the peptide hydrolysis reaction catalyzed by the co-catalytic metallohydrolase from Aeromonas proteolytica (AAP). However, to date, no direct evidence has been reported for the role of Glu-151 during catalytic turnover by AAP. In order to elucidate the catalytic role of Glu-151, altered AAP enzymes have been prepared in which Glu-151 has been substituted with a glutamine, an alanine, and an aspartate. The Michaelis constant (K(m)) does not change upon substitution to aspartate or glutamine, but the rate of the reaction changes drastically in the following order: glutamate (100% activity), aspartate (0.05%), glutamine (0.004%), and alanine (0%). Examination of the pH dependence of the kinetic constants k(cat) and K(m) revealed a change in the pK(a) of a group that ionizes at pH 4.8 in recombinant leucine aminopeptidase (rAAP) to 4.2 for E151D-AAP. The remaining pK(a) values at 5.2, 7.5, and 9.9 do not change. Proton inventory studies indicate that one proton is transferred in the rate-limiting step of the reaction at pH 10.50 for both rAAP and E151D-AAP, but at pH 6.50 two protons and general solvation effects are responsible for the observed effects in the reaction catalyzed by rAAP and E151D-AAP, respectively. Based on these data, Glu-151 is intrinsically involved in the peptide hydrolysis reaction catalyzed by AAP and can be assigned the role of a general acid and base.

  4. Rabbit meat as a source of bacterial foodborne pathogens.

    PubMed

    Rodríguez-Calleja, Jose M; García-López, Isabel; García-López, María-Luisa; Santos, Jesús A; Otero, Andrés

    2006-05-01

    Even though worldwide production of rabbit meat is >1,000,000 tons, little information is available for rabbit meat microbiology. This study provides data on the prevalence of Salmonella, Escherichia coli O157:H7, Yersinia enterocolitica, Listeria spp., motile Aeromonas spp., and Staphylococcus aureus on rabbit meat. A total of 24 rabbit carcasses from two abattoirs and 27 rabbit meat packages from supermarket displays were examined. In addition to culturing methods, associated virulence genes were investigated by PCR in suspect isolates and samples. Neither Salmonella nor E. coli O157:H7 was detected. All samples were negative for virulence-associated invA, stx1, and stx2 genes. At one abattoir, two carcasses (3.9%) carried Y. enterocolitica yst-, and two were positive for the yst gene, although viable Y. enterocolitica cells were not recovered from these samples. Seven samples (13.7%) were contaminated with Listeria. Of them, three were positive for hly and iap genes (Listeria monocytogenes hly+ / iap+), two carried Listeria seeligeri, one carried Listeria ivanovii, and one carried Listeria innocua. For detectable motile Aeromonas spp. (average count, 1.77 +/- 0.62 log CFU/g), the contamination rate was 35.3%, although ca. 90% of the samples were positive for the aerA and/or hlyA genes. The majority of aeromonad isolates were Aeromonas hydrophila aerA+ / hlyA+. Aeromonas caviae, Aeromonas popoffii, Aeromonas schubertii, and the two biovars of Aeromonas veronii were also isolated. The prevalence of S. aureus contamination (average count, 1.37 +/- 0.79 log CFU/g) was 52.9%. Among 27 S. aureus isolates, two harbored genes for staphylococcal enterotoxin B (seb), and two harbored genes for staphylococcal enterotoxin C (sec). The remaining isolates were negative for sea, seb, sec, sed, and see.

  5. Identification of genes associated with copper tolerance in an adhesion-defective mutant of Aeromonas veronii biovar sobria.

    PubMed

    Francki, K T; Chang, B J; Mee, B J; Collignon, P J; Susai, V; Keese, P K

    2000-10-01

    TnphoA mutagenesis was used to identify adhesins of Aeromonas veronii biovar sobria 3767, a strain isolated from a diarrhoeal stool specimen. Six mutants, from a library of 154, exhibited significantly reduced levels of adhesion to HEp-2 cells. Primers to the terminal regions of TnphoA were used for inverse PCR and the product from one mutant was cloned into pBluescript and partial sequence data obtained. Scanning GenBank and EMBL data bases revealed DNA sequence similarity to the copA gene of Pseudomonas syringae pv. tomato which confers resistance to copper and other heavy metals. The transposon was located within the copA gene and the mutant exhibited a reduced tolerance to copper. Primer walking, using the inverse PCR product as a template, revealed three open reading frames (ORFs) copA, B and C in A. veronii biovar sobria 3767. The predicted amino acid sequences of ORFs A and B had significant homology (55 and 34% respectively) to the copA and B proteins of P. syringae. No amino acid or DNA sequence homology existed between ORF C of strain 3767 and any other gene in the data bases scanned. Further analysis of the nucleotide sequence failed to reveal the presence of typical copper regulatory genes within the vicinity of the Aeromonas sequence. The association between copper tolerance and adhesion in A. veronii biovar sobria requires further study.

  6. Neutrophils exert protection in early Aeromonas veronii infections through the clearance of both bacteria and dying macrophages.

    PubMed

    Havixbeck, Jeffrey J; Rieger, Aja M; Churchill, Lucas J; Barreda, Daniel R

    2017-04-01

    Aeromonas veronii is a gram-negative opportunistic pathogen capable of infecting both fish and mammals. Left untreated, natural infection in fish can prove fatal and result in irreparable damage to the aquaculture industry. Neutrophils are essential innate effector cells that play critical roles in pathogen defense. Our aim was to investigate the immunological roles of teleost neutrophils during infection with A. veronii. We began by examining the functional defenses of neutrophils in vitro, where neutrophils efficiently killed the pathogen. In addition, we developed an in vivo infection model to assess the roles of neutrophils during an infection in goldfish. This allowed us to explore the complex dynamics between immune cells and Aeromonas veronii. Interestingly, our studies found that neutrophils are capable of sensing a diverse range of dead and dying cells, resulting in varying downstream responses. Herein, we report that neutrophils internalized dead or dying macrophages previously infected with A. veronii. Moreover, once internalized, neutrophils went on to display classical pro-inflammatory ROS responses, in contrast to the more typical anti-inflammatory responses seen in cells following the uptake of a dead host cell. This led us to hypothesize that during infection, neutrophils are capable of simultaneously clearing dead and dying cells as well as A. veronii. This study provides additional insights into the complex mechanisms by which neutrophils operate within an inflammatory site and contribute to the induction and regulation of acute inflammatory responses.

  7. Biosynthesis of silver nanoparticles using Aeromonas sp. THG-FG1.2 and its antibacterial activity against pathogenic microbes.

    PubMed

    Singh, Hina; Du, Juan; Yi, Tae-Hoo

    2017-05-01

    Silver nanoparticles were prepared through green route with the aid of Aeromonas sp. THG-FG1.2 as reductant. Visual observation, ultraviolet-visible spectroscopy, transmission electron microscopy, elemental mapping, energy dispersive X-ray spectroscopy, selected area diffraction pattern (SAED), and X-ray diffraction (XRD) were used to characterize the synthesized silver nanoparticles. UV visible studies indicated the surface plasmon resonance at 400 nm which depicts the formation of silver nanoparticles. The TEM images show spherical silver nanoparticles of 8-16 nm. XRD and SAED fringes revealed the structure of silver nanoparticles as face centered cubic (fcc). These silver nanoparticles also tested for their antimicrobial potential and showed effective antimicrobial activity against tested pathogens and thus applicable as potent antimicrobial agent. Furthermore, the nanoparticles potential has been reconnoitered for their enhanced synergistic effect with antibiotics against multidrug resistant bacteria. Thus, the silver nanoparticles synthesized by Aeromonas sp. THG-FG1.2, were effective in inhibition of pathogenic microbes and also show enhanced antibacterial activity with antibiotics.

  8. DIFFERENTIATION OF AEROMONAS ISOLATES OBTAINED FROM DRINKING WATER DISTRIBUTION SYSTEM USING MATRIX-ASSISTED LASER DESCRIPTION/IONIZATION-MASS SPECTROMETRY (MALDI-MS)

    EPA Science Inventory

    The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to analyze the...

  9. THE USE OF MATRIX-ASSISTED LASER DESORPTION/IONIZATION-MASS SPECTROMETRY FOR THE IDENTIFICATION OF AEROMONAS ISOLATES OBTAINED FROM WATER DISTRIBUTION SYSTEMS

    EPA Science Inventory

    Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) has long been established as a tool by which microorganisms can be characterized and identified. EPA is investigating the potential of using this technology as a way to rapidly identify Aeromonas species fo...

  10. Draft Genome Sequence of Pseudomonas fluorescens ML11A, an Endogenous Strain from Brook Charr with Antagonistic Properties against Aeromonas salmonicida subsp. salmonicida

    PubMed Central

    Charette, Steve J.; Derome, Nicolas

    2017-01-01

    ABSTRACT Pseudomonas fluorescens ML11A, isolated from brook charr, showed a strong in vitro inhibitory effect against Aeromonas salmonicida subsp. salmonicida, a bacterial fish pathogen. Its genome harbors gene clusters for siderophore and bacteriocin biosynthesis and shares 99% whole-genome identity with P. fluorescens A506, a biological control strain used in agriculture. PMID:28254983

  11. Draft Genome Sequence of Aeromonas caviae CH129, a Marine-Derived Bacterium Isolated from the Coast of São Paulo State, Brazil

    PubMed Central

    Alfonso Vargas, Nadia Catalina; Zimpel, Cristina Kraemer; Pessoa, Adalberto; Rivera, Irma Nelly Gutierrez

    2016-01-01

    We report here the draft genome sequence of Aeromonas caviae CH129, a marine-derived bacterium isolated from the coast of São Paulo state, Brazil. Genomic analysis revealed genes encoding enzymes involved in binding, transport, and chitin metabolism and different virulence-associated factors. PMID:27908996

  12. Draft Genome Sequence of Aeromonas caviae CH129, a Marine-Derived Bacterium Isolated from the Coast of São Paulo State, Brazil.

    PubMed

    Cardozo, Flávio Augusto; Alfonso Vargas, Nadia Catalina; Zimpel, Cristina Kraemer; Pessoa, Adalberto; Rivera, Irma Nelly Gutierrez

    2016-12-01

    We report here the draft genome sequence of Aeromonas caviae CH129, a marine-derived bacterium isolated from the coast of São Paulo state, Brazil. Genomic analysis revealed genes encoding enzymes involved in binding, transport, and chitin metabolism and different virulence-associated factors.

  13. THE IDENTIFICATION OF THE WATER-BORNE PATHOGEN AEROMONAS USING WHOLE CELL ANALYSIS BY MATRIX ASSISTED LASER DESORPTION/IONIZATION-MASS

    EPA Science Inventory

    MALDI-MS has long been established as a tool by which microorganisms can be characterized and identified. The U.S. Environmental Protection Agency (EPA) is investigating the potential of using this technique as a way to rapidly identify Aeromonas species in drinking water. A nu...

  14. Polymorphisms in the umuDC region of Escherichia species. [Escherichia coli; Escherichia alkalescens; Escherichia dispar; Escherichia aurescens

    SciTech Connect

    Sedgwick, S.G.; Robson, M.; Malik, F.

    1988-04-01

    The umuDC operon of Escherichia coli encodes mutagenic DNA repair. The umuDC regions of multiple isolates of E. coli, E. alkalescens, and E. dispar and a single stock of E. aurescens were mapped by nucleotide hybridization. umuDC is located at one end of a conserved tract of restriction endonuclease sites either 12.5 or 14 kilobase pairs long. Rearrangements, including possible deletions, were seen in the polymorphic DNA flanking the conserved tract. Restriction site polymorphisms were not found around the DNA repair gene recA or polA. The junctions of the conserved region contain direct repeats of nucleotide sequences resembling the termini of the Tn3 group of transposons. Possible mechanisms for the generation of these variants are discussed.

  15. ANIMAL ENTEROTOXIGENIC ESCHERICHIA COLI

    PubMed Central

    Dubreuil, J. Daniel; Isaacson, Richard E.; Schifferli, Dieter M.

    2016-01-01

    Enterotoxigenic Escherichia coli (ETEC) is the most common cause of E. coli diarrhea in farm animals. ETEC are characterized by the ability to produce two types of virulence factors; adhesins that promote binding to specific enterocyte receptors for intestinal colonization and enterotoxins responsible for fluid secretion. The best-characterized adhesins are expressed in the context of fimbriae, such as the F4 (also designated K88), F5 (K99), F6 (987P), F17 and F18 fimbriae. Once established in the animal small intestine, ETEC produces enterotoxin(s) that lead to diarrhea. The enterotoxins belong to two major classes; heat-labile toxin that consist of one active and five binding subunits (LT), and heat-stable toxins that are small polypeptides (STa, STb, and EAST1). This chapter describes the disease and pathogenesis of animal ETEC, the corresponding virulence genes and protein products of these bacteria, their regulation and targets in animal hosts, as well as mechanisms of action. Furthermore, vaccines, inhibitors, probiotics and the identification of potential new targets identified by genomics are presented in the context of animal ETEC. PMID:27735786

  16. Selective culturing and genus-specific PCR detection for identification of Aeromonas in tissue samples to assist the medico-legal diagnosis of death by drowning.

    PubMed

    Huys, Geert; Coopman, Vera; Van Varenbergh, Dirk; Cordonnier, Jan

    2012-09-10

    The detection of autochthonous aquatic bacteria in tissue samples from drowning cases is increasingly considered as an alternative approach to assist the medico-legal diagnosis of death by drowning. Bacteria belonging to the genus Aeromonas may be suitable candidates for this application as they are ubiquitous in natural aquatic environments but are generally not part of the human microbiota. The research aims of this study were (i) to develop a sensitive, specific and rapid screening and confirmation method for Aeromonas species in tissue samples and (ii) to evaluate aseptic sternal puncture as a post-mortem sample technique and bone marrow as an alternative matrix to provide evidence of death by drowning. The presence of Aeromonas in tissue samples was verified by cultivation using the selective media Ampicillin Dextrin Agar (ADA) and Ryan's Aeromonas Medium. The use of ADA medium was found most optimal for the sensitive, inexpensive and quick detection of aeromonads in human tissue samples. Positive culture plates were confirmed by harvesting all colonies for DNA extraction and subsequent PCR amplification using Aeromonas genus-specific primers. Aeromonads were detected in lung swab, blood and bone marrow of drowned bodies (n=3), but were negative in these three matrices for all negative controls (n=90) tested. Bone marrow proved to be a suitable alternative matrix and can be sampled post-mortem by an aseptic sternal puncture. In conclusion, this study confirms previous indications that aeromonads in cultures from blood of water bodies can be considered a potential marker for drowning. Given the fact that the number of immersed bodies (drowned and non-drowned) included in this study is statistically not significant, however, more tissue samples need to be investigated to confirm the validity of these methods to aid the diagnosis of death by wet drowning.

  17. L-allo-threonine aldolase from Aeromonas jandaei DK-39: gene cloning, nucleotide sequencing, and identification of the pyridoxal 5'-phosphate-binding lysine residue by site-directed mutagenesis.

    PubMed Central

    Liu, J Q; Dairi, T; Kataoka, M; Shimizu, S; Yamada, H

    1997-01-01

    We have isolated the gene encoding L-allo-threonine aldolase (L-allo-TA) from Aeromonas jandaei DK-39, a pyridoxal 5'-phosphate (PLP)-dependent enzyme that stereospecifically catalyzes the interconversion of L-allo-threonine and glycine. The gene contains an open reading frame consisting of 1,014 nucleotides corresponding to 338 amino acid residues. The protein molecular weight was estimated to be 36,294, which is in good agreement with the subunit molecular weight of the enzyme determined by polyacrylamide gel electrophoresis. The enzyme was overexpressed in recombinant Escherichia coli cells and purified to homogeneity by one hydrophobic column chromatography step. The predicted amino acid sequence showed no significant similarity to those of the currently known PLP-dependent enzymes but displayed 40 and 41% identity with those of the hypothetical GLY1 protein of Saccharomyces cerevisiae and the GLY1-like protein of Caenorhabditis elegans, respectively. Accordingly, L-allo-TA might represent a new type of PLP-dependent enzyme. To determine the PLP-binding site of the enzyme, all of the three conserved lysine residues of L-allo-TA were replaced by alanine by site-directed mutagenesis. The purified mutant enzymes, K51A and K224A, showed properties similar to those of the wild type, while the mutant enzyme K199A was catalytically inactive, with corresponding disappearance of the absorption maximum at 420 nm. Thus, Lys199 of L-allo-TA probably functions as an essential catalytic residue forming an internal Schiff base with PLP of the enzyme to catalyze the reversible aldol reaction. PMID:9171400

  18. Antibacterial activity of guava (Psidium guajava L.) and Neem (Azadirachta indica A. Juss.) extracts against foodborne pathogens and spoilage bacteria.

    PubMed

    Mahfuzul Hoque, M D; Bari, M L; Inatsu, Y; Juneja, Vijay K; Kawamoto, S

    2007-01-01

    The antibacterial activity of guava (Psidium guajava) and neem (Azadirachta indica) extracts against 21 strains of foodborne pathogens were determined--Listeria monocytogenes (five strains), Staphylococcus aureus (four strains), Escherichia coli O157:H7 (six strains), Salmonella Enteritidis (four strains), Vibrio parahaemolyticus, and Bacillus cereus, and five food spoilage bacteria: Pseudomonas aeroginosa, P. putida, Alcaligenes faecalis, and Aeromonas hydrophila (two strains). Guava and neem extracts showed higher antimicrobial activity against Gram-positive bacteria compared to Gram-negative bacteria except for V. parahaemolyticus, P. aeroginosa, and A. hydrophila. None of the extracts showed antimicrobial activity against E. coli O157:H7 and Salmonella Enteritidis. The minimum inhibitory concentration (MIC) of ethanol extracts of guava showed the highest inhibition for L. monocytogenes JCM 7676 (0.1 mg/mL), S. aureus JCM 2151 (0.1 mg/mL), S. aureus JCM 2179 (0.1 mg/mL), and V. parahaemolyticus IFO 12711 (0.1 mg/mL) and the lowest inhibition for Alcaligenes faecalis IFO 12669, Aeromonas hydrophila NFRI 8282 (4.0 mg/mL), and A. hydrophila NFRI 8283 (4.0 mg/mL). The MIC of chloroform extracts of neem showed similar inhibition for L. monocytogenes ATCC 43256 (4.0 mg/mL) and L. monocytogenes ATCC 49594 (5.0 mg/mL). However, ethanol extracts of neem showed higher inhibition for S. aureus JCM 2151 (4.5 mg/mL) and S. aureus IFO 13276 (4.5 mg/mL) and the lower inhibition for other microorganisms (6.5 mg/mL). No significant effects of temperature and pH were found on guava and neem extracts against cocktails of L. monocytogenes and S. aureus. The results of the present study suggest that guava and neem extracts possess compounds containing antibacterial properties that can potentially be useful to control foodborne pathogens and spoilage organisms.

  19. Exposure to pairs of Aeromonas strains enhances virulence in the Caenorhabditis elegans infection model

    PubMed Central

    Mosser, Thomas; Talagrand-Reboul, Emilie; Colston, Sophie M.; Graf, Joerg; Figueras, Maria J.; Jumas-Bilak, Estelle; Lamy, Brigitte

    2015-01-01

    Aeromonad virulence remains poorly understood, and is difficult to predict from strain characteristics. In addition, infections are often polymicrobial (i.e., are mixed infections), and 5–10% of such infections include two distinct aeromonads, which has an unknown impact on virulence. In this work, we studied the virulence of aeromonads recovered from human mixed infections. We tested them individually and in association with other strains with the aim of improving our understanding of aeromonosis. Twelve strains that were recovered in pairs from six mixed infections were tested in a virulence model of the worm Caenorhabditis elegans. Nine isolates were weak worm killers (median time to death, TD50, ≥7 days) when administered alone. Two pairs showed enhanced virulence, as indicated by a significantly shortened TD50 after co-infection vs. infection with a single strain. Enhanced virulence was also observed for five of the 14 additional experimental pairs, and each of these pairs included one strain from a natural synergistic pair. These experiments indicated that synergistic effects were frequent and were limited to pairs that were composed of strains belonging to different species. The genome content of virulence-associated genes failed to explain virulence synergy, although some virulence-associated genes that were present in some strains were absent from their companion strain (e.g., T3SS). The synergy observed in virulence when two Aeromonas isolates were co-infected stresses the idea that consideration should be given to the fact that infection does not depend only on single strain virulence but is instead the result of a more complex interaction between the microbes involved, the host and the environment. These results are of interest for other diseases in which mixed infections are likely and in particular for water-borne diseases (e.g., legionellosis, vibriosis), in which pathogens may display enhanced virulence in the presence of the right partner. This

  20. Diversity and Homogeneity among Small Plasmids of Aeromonas salmonicida subsp. salmonicida Linked with Geographical Origin

    PubMed Central

    Attéré, Sabrina A.; Vincent, Antony T.; Trudel, Mélanie V.; Chanut, Romain; Charette, Steve J.

    2015-01-01

    Furunculosis, which is caused by Aeromonas salmonicida subsp. salmonicida, is a major salmonid disease in fish farms worldwide. Several plasmids found in this bacterium confer phenotypes such drug resistance and virulence. Small plasmids (pAsa1, pAsa2, pAsa3, and pAsal1) related to ColE1- and ColE2-type replicons are usually present in its normal plasmidome. In the present study, with the objective to investigate if these plasmids display particularities related to the origin of the isolates bearing them, a total of 153 isolates, including 78 new and 75 previously described, were analyzed for the presence of small plasmids by PCR and DNA restriction fragment profiling. A geographical dichotomy between Canadian and European isolates for their propensity to do not have pAsa3 or pAsal1 was found. In addition, the genotyping analysis led to the identification of two European isolates harboring an unusual pAsal1. An investigation by next-generation sequencing (NGS) of these two isolates shed light on two pAsal1 variants (pAsal1C and pAsal1D). As with pAsal1B, another pAsal1 variant previously described, these two new variants bore a second insertion sequence (ISAS5) in addition to the usual ISAS11. The characterization of these variants suggested that they could predominate over the wild-type pAsal1 in stressful conditions such as growth at temperatures of 25°C and above. To obtain a comprehensive portrait of the mutational pressure on small plasmids, 26 isolates whose DNA had been sequenced by NGS were investigated. pAsa3 and pAsal1 were more prone to mutations than pAsa1 and pAsa2, especially in the mobA gene, which encodes a relaxase and a primase. Lastly, the average copy number of each plasmid per cell was assessed using raw sequencing data. A clear trend with respect to the relative proportion per cell of each plasmid was identified. Our large-scale study revealed a geographical dichotomy in small plasmid repertoire in addition to a clear trend for pAsa3 and p

  1. QTL detection for Aeromonas salmonicida resistance related traits in turbot (Scophthalmus maximus)

    PubMed Central

    2011-01-01

    Background Interactions between fish and pathogens, that may be harmless under natural conditions, often result in serious diseases in aquaculture systems. This is especially important due to the fact that the strains used in aquaculture are derived from wild strains that may not have had enough time to adapt to new disease pressures. The turbot is one of the most promising European aquaculture species. Furunculosis, caused by the bacterium Aeromonas salmonicida, produces important losses to turbot industry. An appealing solution is to achieve more robust broodstock, which can prevent or diminish the devastating effects of epizooties. Genomics strategies have been developed in turbot to look for candidate genes for resistance to furunculosis and a genetic map with appropriate density to screen for genomic associations has been also constructed. In the present study, a genome scan for QTL affecting resistance and survival to A. salmonicida in four turbot families was carried out. The objectives were to identify consistent QTL using different statistical approaches (linear regression and maximum likelihood) and to locate the tightest associated markers for their application in genetic breeding strategies. Results Significant QTL for resistance were identified by the linear regression method in three linkage groups (LGs 4, 6 and 9) and for survival in two LGs (6 and 9). The maximum likelihood methodology identified QTL in three LGs (5, 6 and 9) for both traits. Significant association between disease traits and genotypes was detected for several markers, some of them explaining up to 17% of the phenotypic variance. We also identified candidate genes located in the detected QTL using data from previously mapped markers. Conclusions Several regions controlling resistance to A. salmonicida in turbot have been detected. The observed concordance between different statistical methods at particular linkage groups gives consistency to our results. The detected associated

  2. Persistence, transmission, and virulence characteristics of Aeromonas strains in a duckweed aquaculture-based hospital sewage water recycling plant in Bangladesh.

    PubMed

    Rahman, Mokhlasur; Huys, Geert; Rahman, Motiur; Albert, M John; Kühn, Inger; Möllby, Roland

    2007-03-01

    The persistence and transmission of Aeromonas in a duckweed aquaculture-based hospital sewage water treatment plant in Bangladesh was studied. A total of 670 samples from different sites of the hospital sewage water treatment plant, from feces of hospitalized children suffering from diarrhea, from environmental control ponds, and from feces of healthy humans were collected over a period of three years. In total, 1,315 presumptive Aeromonas isolates were biochemically typed by the PhenePlate rapid screening system (PhP-AE). A selection of 90 representative isolates was further analyzed with PhenePlate (PhP) extended typing (PhP-48), fatty acid methyl ester analysis, and amplified fragment length polymorphism (AFLP) fingerprinting. In addition, the prevalence of the putative virulence factors hemolysin and cytotoxin and the presence of the cytolytic enterotoxin gene (AHCYTOEN) were analyzed. Aeromonas was found at all sites of the treatment plant, in 40% of the samples from environmental control ponds, in 8.5% of the samples from hospitalized children suffering from diarrhea, and in 3.5% of samples from healthy humans. A significantly high number of Aeromonas bacteria was found in duckweed, which indicates that duckweed may serve as a reservoir for these bacteria. PhP-AE typing allowed identification of more than 192 distinct PhP types, of which 18 major PhP types (MTs) were found in multiple sites and during several occasions. AFLP fingerprinting revealed the prevalence of genotypically indistinguishable Aeromonas isolates among certain PhP MTs recovered from different sampling occasions and/or at multiple sites. Hemolytic and cytotoxic activities were observed in 43% of the tested strains, whereas 29% possessed the cytolytic enterotoxin gene AHCYTOEN. Collectively, two specific MTs associated with diarrhea were shown to exhibit high cytotoxicity. Furthermore, all tested isolates of these major types were positive for the cytolytic enterotoxin gene. In conclusion

  3. Simultaneous Detection of Aeromonas salmonicida, Flavobacterium psychrophilum, and Yersinia ruckeri, Three Major Fish Pathogens, by Multiplex PCR

    PubMed Central

    del Cerro, A.; Marquez, I.; Guijarro, J. A.

    2002-01-01

    A multiplex PCR assay based on the 16S rRNA genes was developed for the simultaneous detection of three major fish pathogens, Aeromonas salmonicida, Flavobacterium psychrophilum, and Yersinia ruckeri. The assay proved to be specific and as sensitive as each single PCR assay, with detection limits in the range of 6, 0.6, and 27 CFU for A. salmonicida, F. psychrophilum, and Y. ruckeri, respectively. The assay was useful for the detection of the bacteria in artificially infected fish as well as in fish farm outbreaks. Results revealed that this multiplex PCR system permits a specific, sensitive, reproducible, and rapid method for the routine laboratory diagnosis of infections produced by these three bacteria. PMID:12324372

  4. Aeromonas veronii, a tributyltin (TBT)-degrading bacterium isolated from an estuarine environment, Ria de Aveiro in Portugal.

    PubMed

    Cruz, Andreia; Caetano, Tânia; Suzuki, Satoru; Mendo, Sónia

    2007-12-01

    Organotin compounds are used in a variety of industrial processes therefore their subsequent discharge into the environment is widespread. Bacteria play an important role in biogeochemical transformations acting as natural decontamination agents. Therefore, screening for tributyltin (TBT)-resistant and -degrading bacteria is relevant for the selection of isolates with decontamination ability of these polluted areas. With this purpose, 50 strains were isolated from sediment and water from Ria de Aveiro and their tolerance to TBT, up to 3mM, was evaluated. Generally, occurrence of highly TBT-resistant bacteria was observed, and Gram negative bacteria exhibited more tolerance to TBT than Gram positive bacteria. A memory response was observed when bacteria were progressively exposed to increasingly higher TBT concentrations. One isolate, Aeromonas veronii Av27, highly resistant to TBT (3mM) uses this compound as carbon source and degrades it to less toxic compounds.

  5. In vitro antibacterial activity of chitosan and chitosan oligosaccharide lactate against important gram negative warmwater fish pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aim: The antibacterial activities of chitosan (CS) and its derivative chitosan oligosaccharide lactate (COL) were evaluated against Aeromonas hydrophila, Edwardsiella ictaluri and Flavobacterium columnare, three highly pathogenic bacteria of warmwater finfish. Methods and Results: The kinetics of ce...

  6. Identification of iron and heme utilization genes in Aeromonas and their role in the colonization of the leech digestive tract

    PubMed Central

    Maltz, Michele; LeVarge, Barbara L.; Graf, Joerg

    2015-01-01

    It is known that many pathogens produce high-affinity iron uptake systems like siderophores and/or proteins for utilizing iron bound to heme-containing molecules, which facilitate iron-acquisition inside a host. In mutualistic digestive-tract associations, iron uptake systems have not been as well studied. We investigated the importance of two iron utilization systems within the beneficial digestive-tract association Aeromonas veronii and the medicinal leech, Hirudo verbana. Siderophores were detected in A. veronii using chrome azurol S. Using a mini Tn5, a transposon insertion in viuB generated a mutant unable to utilize iron using siderophores. The A. veronii genome was then searched for genes potentially involved in iron utilization bound to heme-containing molecules. A putative outer membrane heme receptor (hgpB) was identified with a transcriptional activator, termed hgpR, downstream. The hgpB gene was interrupted with an antibiotic resistance cassette in both the parent strain and the viuB mutant, yielding an hgpB mutant and a mutant with both iron uptake systems inactivated. In vitro assays indicated that hgpB is involved in utilizing iron bound to heme and that both iron utilization systems are important for A. veronii to grow in blood. In vivo colonization assays revealed that the ability to acquire iron from heme-containing molecules is critical for A. veronii to colonize the leech gut. Since iron and specifically heme utilization is important in this mutualistic relationship and has a potential role in virulence factor of other organisms, genomes from different Aeromonas strains (both clinical and environmental) were queried with iron utilization genes of A. veronii. This analysis revealed that in contrast to the siderophore utilization genes heme utilization genes are widely distributed among aeromonads. The importance of heme utilization in the colonization of the leech further confirms that symbiotic and pathogenic relationships possess similar

  7. Aeromonas sobria sepsis complicated by rhabdomyolysis in an HIV-positive patient: case report and evaluation of traits associated with bacterial virulence.

    PubMed

    Stano, Francesca; Brindicci, Gaetano; Monno, Rosa; Rizzo, Caterina; Ghezzani, Francesca; Carbonara, Sergio; Guaglianone, Emilio; Donelli, Gianfranco; Monno, Laura

    2009-05-01

    Human infection with Aeromonas species is uncommon and most often due to trauma with exposure to contaminated water or soil. A 43-year-old HIV- and hepatitis C virus (HCV)-infected male, after a two-week course of corticosteroid therapy for an autoimmune anemia, developed diarrhea, dermatologic manifestations and a multiple organ dysfunction syndrome, resulting in death. Although stool samples were repeatedly negative, two sets of blood cultures obtained during a single peak of fever yielded the post-mortem isolation of a Gram-negative, oxidase-positive, beta-hemolytic bacillus that was identified as Aeromonas sobria. Empiric antibiotic therapy was unsuccessful. Evaluation of the virulence-associated traits of the clinical isolate (adhesion, cytotoxicity activity, biofilm production) showed that the strain was a poor producer of recognized virulence factors, thereby indicating that the unfortunate coexistence of HIV infection, HCV-related liver cirrhosis and corticosteroids played a key role in the clinical course.

  8. Reclassification of Aeromonas sharmana to a new genus as Pseudaeromonas sharmana gen. nov., comb. nov and description of Pseudaeromonas pectinilytica sp. nov. isolated from a fresh water stream.

    PubMed

    Padakandla, Shalem Raj; Chae, Jong-Chan

    2016-12-14

    A Gram-negative, rod shaped, facultatively anaerobic, motile bacterium strain AR1T was isolated from a fresh water stream in Jeonju, South Korea. Strain AR1T showed highest 16S rRNA gene sequence similarity (96.83%) and also formed a separate clade with Aeromonas sharmana GPTSA-6T in the phylogenetic tree constructed among the members of the family Aeromonadaceae. Major cellular fatty acids are summed feature 3 (C16:1ω7C/C16:1ω6C) and C16:0. Diphosphatidylglycerol, diphosphatidylethanolamine and phosphatidylglycerol are the predominant polar lipids. Genomic DNA G+C content was found to be 54.7 mol %. However earlier studies on 16S rRNA, gyrB, rpoD and universal target region of cpn60 gene sequences of the members of genus Aeromonas recommended the transfer of Aeromonas sharmana to a new genus. Hence, based on the comparative polyphasic data obtained during the present study and also on the previous recommendations, it is proposed that Aeromonas sharmana be transferred to a novel genus as Pseudaeromonas sharmana gen. nov. comb. nov. with strain GPTSA-6T (=DSM 17445T=MTCC 7090T=CIP 109378T=CCUG 54939T) as the type species of the genus. Also, it is proposed that the yet unknown bacterium strain AR1T be designated as a species of this genus namely Pseudaeromonas pectinilytica gen. nov. sp. nov. The type strain is AR1T (=KCTC 42754T =JCM 31503T).

  9. The development of a matrix-assisted laser desorption/ionization mass spectrometry-based method for the protein fingerprinting and identification of Aeromonas species using whole cells.

    PubMed

    Donohue, Maura J; Smallwood, Anthony W; Pfaller, Stacy; Rodgers, Mark; Shoemaker, Jody A

    2006-06-01

    This report describes the development of a method to detect the waterborne pathogen Aeromonas using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The genus Aeromonas is one of several medically significant