Phenotypic, serological and genetic characterization of Pseudomonas anguilliseptica strains isolated from cod, Gadus morhua L., in northern Europe (original) (raw)
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Detection of Virulence factors of Pseudomonas species isolated from fresh water fish by PCR
Benha Veterinary Medical Journal, 2016
This study was conducted on 100 diseased Nile tilapia (O. niloticus) fish of various sizes collected from different fish markets in Kaliobia Governorate to estimate the prevalence of Pseudomonas infection and detection of some virulence genes in the isolated P. aeruginosa strains. The results of bacteriological examination revealed that the prevalence of Pseudomonas septicemia with Pseudomonas species isolation was 17.0% (17 \ 100 examined fish). These cases were attributed to P. anguilliseptica; P. aeruginosa and P. fluorescens (14/43.7%; 12/37.5% and 6/18.8%), respectively. In addition, 32 Pseudomonas species were isolated, 11from liver samples (34.4%); 10 from kidney samples (31.2%); 6 from gill samples (18.8%) and 5 from skin samples (15.6%). Moreover, 14 P. anguilliseptica were isolated with an incidence of 35.7%, 28.6%, 21.4% and 14.3% followed by 12 P. aeruginosa 33.3%, 25%,16.7% and 25% respectively; 6 P. fluorescens 33.3%; 50.0%,16.7% and 0.0% from the liver, kidney, gill and skin samples respectively. The in-vitro antimicrobial sensitivity test showed that the isolated Pseudomonas strains were sensitive to gentamycin; enrofloxacin; norfloxacin; ciprofloxacin and florphenicol. Meanwhile; they were intermediate sensitive for doxycycline; sulfa-trimethoprim; oxytetracycline; nalidixic acid and streptomycin. In contrast, they were resistant for cefotaxime; erythromycin; amoxicillin; methicillin; oxacillin and ampicillin. Moreover, the PCR results revealed that, opr L and exo S virulence genes were detected in all six studied strains (100.0%). Meanwhile, phz M virulence gene was detected in 5 out of 6 studied strains (83.3%) and tox A virulence gene was detected in 4 out of 6 studied strains (66.7%) i.e., all studied strains were Ps. aeruginosa and all of them were virulent strains.
Diseases of Aquatic Organisms, 2002
A PCR-based detection system for Pseudomonas anguilliseptica was evaluated. The primer combination PAF-PAR (forward primer PAF = 5'-GACCTCGCCATTA-3', reverse primer PAR = 5'-CTCAGCAGTTTTGAAAG-3') gave a unique and specific amplification product of 439 bp at an annealing temperature of 46°C with all the P. anguilliseptica isolates and strains (n = 56) but no amplification products were observed with any other Pseudomonas species or phylogenetically related bacteria tested. The PCR assay had a detection limit of 170 to 200 cells per PCR tube, which was improved 8-fold when the PCR amplification product was used as a nonradioactive probe in blotting hybridization experiments. The PCR assay allowed the specific and reliable detection of P. anguilliseptica within 8 h, compared with up to 10 d required for its isolation and further characterization by conventional microbiological approaches. Clinical isolates of P. anguilliseptica recovered from several winter disease (WD) outbreaks diagnosed in sea bream Sparus aurata in Spain and Portugal between 1996 and 2001 were characterized by pulse field-gel electrophoresis (PFGE) macrorestriction analysis. The 54 clinical isolates analyzed were included in 4 different pulsotypes. Pulsotypes B and C represented 54 and 25% of the isolates, respectively, and were responsible for most of the WD outbreaks diagnosed in Spain between 1996 and 2001. The implication of asymptomatic infected carriers in the dissemination and spread of WD is discussed.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2012
Five Gram-negative bacterial isolates, recovered from an outbreak that occurred in March 2006 in Huelva, Spain, affecting adult diseased cultured wedge sole [Dicologlossa cuneata (Moreau)], were characterized phenotypically and genotypically in order to clarify their taxonomic position. On the basis of 16S rRNA gene sequence analysis, the isolates were included in the genus Pseudomonas , within the Pseudomonas fluorescens -related species group, their closest relatives being the Pseudomonas jessenii and Pseudomonas koreensis subgroups. The highest sequence similarities were recorded with the type strains of Pseudomonas reinekei , P. moorei , P. umsongensis , P. jessenii and P. mohnii (99.4–99.3 % similarity). Sequence analysis of the housekeeping genes gyrB and rpoD clearly differentiated the isolates from currently described Pseudomonas species, the highest sequence similarities recorded to type strains being below 95 % for both genes. Phylogenetic analysis using concatenated seque...
Phenotypic and Genotypic Comparison of Pseudomonas stutzeri in Freshwater Fish in Indonesia
Journal of Agricultural Science and Technology B, 2015
Pseudomonas stutzeri caused an outbreak of freshwater fish in Luwuk Banggai (tilapia and catfish), Bali (tilapia), Jambi (tilapia and catfish) and Tanjung Pinang (catfish). The study was purposed to comprehensively identify special phenotypic and genotypic characteristics of P. stutzeri isolated from several areas in Indonesia, including its morphometric and biochemical characteristics and molecular variation. Bacteria were isolated from internal organs (kidney, ulcer and eye) of fish. They were then identified using morphology and biochemical test. DNA isolates were entirely extracted, amplified and reversed on 16S rRNA region, and further then were sequenced. Phylogenetic trees of bacteria were constructed using neighbor-joining and maximum-parsimony methods. The colony were similar, such as rod shape (Jambi, Tanjung Pinang, Bali), bacil shape (Luwuk Banggai), transparant in tryptic soy agar (TSA) (Luwuk Banggai), creamy beige in glutamate starch phenol red (GSP) (Bali), gram negative, motile, no reaction in the oxidative-fermentative test, positive result in catalase and oxidase test, negative in lysine decarboxylase and ornithine decarboxylase test and positive result in indole test; gelatin was degraded (only Bali), urea was not degraded, no color change in Methyl-red and Voges-proskaeur (MR-VP) test; acid not produce from glucose, inositol or sucrose. Citrate was utilized by some isolates: positive (Jambi, Tanjung Pinang) and negative (Bali, Luwuk Banggai). Results showed us that isolates of Jambi, Bali and Tanjung Pinang were monophyletic species with P. stutzeri S8 and ZH-1 comparing to gen bank. However, merely phenotypic analysis among Pseudomonas sp. was confused compared to each other.
Oreochromis niloticus from Qaroun and El Rayan lakes were collected and examined for the presence of pseudomonas species. The organisms were found in 30.83% of the 480 examined fish with some fish, especially during the episode of mass mortality showing, typical signs of pseudomonas septicemia; redness all over the body, abdominal swelling, eyes cloudness, scales detachment and congested gills. Culture from liver, spleen, kidneys and gills on pseudomonas agar media yielded different species of pseudomonas with morphological and biochemical characters identical to P. fluorescens biovar I, II, III, P. anguilliseptica, P. putida and P. aureginosa isolated from the same fish and other fish species of Qaroun and El Rayan lakes as well as for those recovered from other kinds of fish. The prevalence of infection revealed significant difference among four batches, it was 43.and the organisms were mainly isolated from liver and kidney (35 and 30%, respectively). Challenge tests revealed that P. angulliseptica was the highest pathogenic one and induced 96.66% mortality, while P. fluroscens injected groups showed 2% mortality. Antibiogram sensituity revealed high sensitivity to Avatryl and Amikicin and sensitivity to Gentamicin, Erythromycin, Novobiocin and Sulfa-trimethoprime. Three major proteins of 29 KDa, 30 KDa and 35 KDa in the whole cell proteins of pseudomonas strains. The pathogenic strains P. aeruginosa, P. putida and P. angulliseptica had 9-11 protein bands ranged from 23.4 to 100.05 KDa in molecular weight, while non-pathogenic strains; P. fluroscens biovar I, II and III showed 7-9 protein bands and of molecular weight ranged from 22 to 87 KDa. It was concluded that different pseudomonas diseases were considered Qaroun and El-Rayan lakes causes septicemia in O. niloticus and the pathogenic species include P. anguilliseptica, P. putida and P. aureginosa. Avatryl and Amikicin are the best drug for treatment of septicemia caused by the above mentioned species in O. niloticus.
Fisheries and aquatic sciences, 2010
In recent years, several southern coastal fish farms in Korea have experienced 2-30% mortality in striped beakperch suffering from bacterial infections during the spring. In this study, we identified a bacterium isolated from diseased striped beakperch as Pseudomonas anguilliseptica via a biochemical test and 16S rDNA sequence analysis. To evaluate the susceptibility of striped beakperch to P. anguilliseptica, 4.39×10 7 or 4.39×10 5 CFU/fish of bacteria were injected intraperitoneally at 18 ± 1°C into fish weighing 5.5 g. Cumulative mortalities reached 100% and 45% in the 4.39×10 7 and 4.39×10 5 CFU/fish infected groups, respectively. Experimentally infected fish showed cell-associated inflammation as well as bacteria in the kidneys and spleen. This study is the first report of striped beakperch mortality caused by P. anguilliseptica, which has pathogenicity in striped beakperch.
Microbial pathogenesis, 2018
The present study was investigating the clinical pictures, prevalence, as well as the ecological conditions associated with Pseudomonas anguilliseptica outbreaks in four cultured seabream, Sparus aurata farms at different localities in Egypt during winter of 2016. The phenotypic and genotypic patterns of Pseudomonas isolates were investigated. The existence of intraspecific heterogeneity among different isolates was analyzed using Restriction Fragment Length Polymorphism (RFLP) technique. Attempts on disease control using antibiogram or dietary supplement were also considered. To achieve these goals, various commercial antibiotic discs were analyzed against Ps. anguilliseptica isolates using the disc diffusion method. Additionally, the impact of one-month dietary incorporation with 3% garlic extract or 0.5% potassium diformate on S. aurata viability and response for prolonged bathing treatment with florfenicol was evaluated following challenge with the virulent strain of Ps. anguill...
Applied and Environmental Microbiology, 2002
The population dynamics of pseudomonads in gilt-head sea bream Mediterranean fish (Sparus aurata) stored under different conditions were studied. Phenotypic analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins were performed to identify a total of 106 Pseudomonas strains isolated from S. aurata stored under different temperatures (at 0, 10, and 20°C) and packaging conditions (air and a modified atmosphere of 40% CO 2 -30% N 2 -30% O 2 ). Pseudomonas lundensis was the predominant species, followed by Pseudomonas fluorescens, while Pseudomonas fragi and Pseudomonas putida were detected less frequently. Fluorescent Pseudomonas strains dominated under air conditions, while proteolytic and less lipolytic strains dominated under modified-atmosphere packaging. Different storage conditions appear to govern the selection of pseudomonads in gilt-head sea bream fish.