scholarly journals Species-Specific Duplex PCR for Detecting the Important Fish Pathogens Vibrio anguillarum and Edwardsiella tarda

2013 ◽  
Vol 16 (4) ◽  
pp. 273-277 ◽  
Author(s):  
Geon-A Jo ◽  
Sae-Bom Kwon ◽  
Na-Kyeong Kim ◽  
Muhammad Tofazzal Hossain ◽  
Yu-Ri Kim ◽  
...  
Keyword(s):  
Vibrio Anguillarum ◽  
Edwardsiella Tarda ◽  
Duplex Pcr ◽  
Fish Pathogens ◽  
Species Specific ◽  
Important Fish

scholarly journals Commercial phenotypic tests (API 20E) in diagnosis of fish bacteria: a review

2008 ◽  
Vol 52 (No. 2) ◽  
pp. 49-53 ◽  
Author(s):  
N. Topic Popovic ◽  
R. Coz-Rakovac ◽  
I. Strunjak-Perovic
Keyword(s):  
Vibrio Anguillarum ◽  
Rapid Identification ◽  
Identification System ◽  
Bacterial Isolates ◽  
Yersinia Ruckeri ◽  
Fish Health ◽  
Fish Pathogens ◽  
Phenotypic Tests ◽  
Fish Bacteria ◽  
Important Fish

The available data concerning rapid identification of fish bacteria via commercial phenotypic tests demonstrate that there is no agreement regarding the choice of the tests. However, API 20E, an identification system for Enterobacteriaceae and other non-fastidious Gram-negative rods developed for clinical specimens, seems to be increasingly used for the identification of fish pathogens. In this review, adaptation of API 20E for fish bacterial isolates and its distinctiveness for fish bacteria was assessed. Some strains are wrongly identified because they are not included in the database of API 20E system. API 20E reactions should be compared with the diagnostic schemes based on reactions in conventional phenotypic tests. Due to their significance for fish health and impact on the aquaculture, and because of the need for their rapid identification, some important fish bacteria should be included in the API 20E system, such as <i>Yersinia ruckeri</i>, <i>Edwardsiella ictaluri</i>, <i>Vibrio anguillarum</i>.

Identification of Pathogenic Fish Bacteria Using the API ZYM System

1993 ◽  
Vol 50 (6) ◽  
pp. 1137-1141 ◽  
Author(s):  
Masahiro Sakai ◽  
M. K. Soliman ◽  
T. Yoshida ◽  
Masanori Kobayashi
Keyword(s):  
Bacterial Species ◽  
Vibrio Anguillarum ◽  
Aeromonas Salmonicida ◽  
Edwardsiella Tarda ◽  
Gram Stain ◽  
Fish Pathogen ◽  
Fish Pathogens ◽  
Renibacterium Salmoninarum ◽  
Hemolytic Streptococcus ◽  
Fish Bacteria

The API ZYM system was used to identify the bacterial fish pathogens Enterococcus seriolicida, β-hemolytic Streptococcus sp., Renibacterium salmoninarum, Aeromonas salmonicida, A. hydrophila, Pasteurella piscicida, Edwardsiella tarda, Vibrio anguillarum, Pseudomonas fluorescens, Flexibacter columnaris, and F. maritimus. As additional tests, Gram stain and the activities of cytochrome oxidase and catalase were examined. The results of APS ZYM and the additional tests were coded for easy identification. Several biotypes were demonstrated by Streptococcus sp., V. anguillarum, and A. hydrophila. Other bacteria showed uniform profiles. This system can distinguish each fish pathogen from all other bacterial species examined.

Ultraviolet Treatment of Water for Destruction of Five Gram-Negative Bacteria Pathogenic to Fishes

1977 ◽  
Vol 34 (8) ◽  
pp. 1244-1249 ◽  
Author(s):  
G. L. Bullock ◽  
H. M. Stuckey
Keyword(s):  
Particulate Matter ◽  
Ultraviolet Irradiation ◽  
Vibrio Anguillarum ◽  
Spring Water ◽  
Aeromonas Salmonicida ◽  
Water Disinfection ◽  
Gram Negative Bacteria ◽  
Fish Pathogens ◽  
Gram Negative ◽  
Ultraviolet Treatment

Filtration (25 nm) and ultraviolet irradiation dosages of 13,100–29,400 microwatt seconds per square centimetre (μW∙s∙cm−2) effected a 99.98–100% reduction of five gram-negative fish pathogens — Aeromonas salmonicida, A. hydrophila, Vibrio anguillarum, Pseudomonas fluorescens, and the enteric redmouth organism in 12.5 °C clear spring water or spring water containing particulate matter. Filtration and a dosage of 4500 μW∙s∙cm−2 killed 99.83–100% of test strains in spring water and 4000–4750 μW∙s∙cm−2 killed 99.33–99.99% in water with particulate matter. Irradiation of unfiltered water containing particulate matter was less effective, especially at dosages of 5000 μW∙s∙cm−2 or less, which killed 97–99.94% of strains. Filtration and 13,100 μW∙s∙cm−2 irradiation of water containing A. salmonicida prevented transmission of furunculosis. Key words: ultraviolet irradiation, bacterial fish pathogens, water disinfection

Species-specific duplex PCR for the detection of Pratylenchus penetrans

Nematology ◽  
2009 ◽  
Vol 11 (6) ◽  
pp. 847-857 ◽  
Author(s):  
Lieven Waeyenberge ◽  
Nicole Viaene ◽  
Maurice Moens
Keyword(s):  
Internal Control ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Duplex Pcr ◽  
Specific Primers ◽  
5.8S Rrna Gene ◽  
5.8S Rrna ◽  
Wide Range ◽  
Pcr Products ◽  
Species Specific

Abstract ITS1, the 5.8S rRNA gene and ITS2 of the rDNA region were sequenced from 20 different Pratylenchus species. Additionally, the same region was sequenced from seven populations of P. penetrans. After purifying, cloning and sequencing the PCR products, all sequences were aligned in order to find unique sites suitable for the design of species-specific primers for P. penetrans. Since ITS regions showed variability between and even within populations of P. penetrans, only three small DNA sequences were suitable for the construction of three potentially useful species-specific primers. New species-specific primers were paired with existing universal ITS primers and tested in all possible primer combinations. The best performing primer set, supplemented with a universal 28S rDNA primer set that served as an internal control, was tested in duplex PCR. The ideal annealing temperature, Mg2+ concentration and primer ratios were then determined for the most promising primer set. The optimised duplex PCR was subsequently tested on a wide range of different Pratylenchus spp. and 25 P. penetrans populations originating from all over the world. To test the sensitivity, the duplex PCR was conducted on DNA extracted from a single P. penetrans nematode mixed with varying amounts of nematodes belonging to another Pratylenchus species. Results showed that a reliable and sensitive P. penetrans species-specific duplex PCR was constructed.

Molecular characterization of Vibrio anguillarum biotype 2

1981 ◽  
Vol 27 (10) ◽  
pp. 1011-1018 ◽  
Author(s):  
Michael H. Schiewe ◽  
Jorge H. Crosa
Keyword(s):  
Vibrio Anguillarum ◽  
Virulence Plasmid ◽  
Multicopy Plasmid ◽  
Nick Translation ◽  
Fish Pathogens ◽  
Mole Percent ◽  
Discrete Nature ◽  
Sequence Relationships ◽  
Restriction Endonuclease Cleavage

Examination of polynucleotide sequence relationships among 11 strains of Vibrio anguillarum biotype 2 isolated from moribund fish in North America and Japan demonstrated the highly conserved character of this group of fish pathogens and, moreover, confirmed its discrete nature from V. anguillarum biotype 1. Additional molecular analyses of the V. anguillarum biotype 2 strains revealed the universal presence of a multicopy plasmid with a molecular mass of approximately 20 × 106 daltons and a mole percent guanine plus cytosine of 44. The plasmid of strain DF3K was representative of this molecular species and was designated pMJ101. Subsequent DNA–DNA hybridizations using nick translation-labeled pMJ101 as a probe indicated all the 20 × 106 dalton plasmids were either identical or highly conserved, and, furthermore, that pMJ101 was apparently unrelated to either the virulence plasmid, pJM1, of V. anguillarum biotype 1 or to representatives of common plasmid incompatibility groups. The lack of relatedness between pMJ101 and pJM1 was further supported by differences in their restriction endonuclease cleavage patterns.

scholarly journals Pathology of Edwardsiella tarda infection in African catfish, Clarias gariepinus (Burchell 1822), fingerlings

2015 ◽  
Vol 23 (3) ◽  
pp. 141-148 ◽  
Author(s):  
Thangapalam Jawahar Abraham ◽  
Prakash Kumar Mallick ◽  
Harresh Adikesavalu ◽  
Sayani Banerjee
Keyword(s):  
Inflammatory Responses ◽  
Clarias Gariepinus ◽  
Edwardsiella Tarda ◽  
Phenotypic Characterization ◽  
African Catfish ◽  
Lymphocyte Infiltration ◽  
Hematopoietic Tissue ◽  
Fish Pathogens ◽  
Mucus Production ◽  
Focal Necrosis

AbstractEdwardsiella tarda is one of the serious fish pathogens infecting both cultured and wild fish species. This study aimed to assess the phenotypic characterization and pathogenicity of E. tarda isolated from Clarias gariepinus (Burchell) with dropsy and histopathological alterations. The causative agent was identified with Vitek 2, and its pathogenicity was determined by intramuscular injection. The challenged catfish exhibited vertical hanging, frothing, excess mucus production, listing, swollen abdomen, anorexia, fin and tail rot, and reddish operculum. The LD50of E. tarda PBB and PBP strains was found to be 8.52 × 106and 1.68 × 107cells fish-1, respectively. Histopathological observations on catfish infected naturally revealed lymphocyte infiltration in muscle and focal necrosis, hyperplasia, edema, and swelling of the gill lamellar epithelium. The kidney of diseased fish exhibited ischemic type tubulopathy, necrosis of nephritic tubules, hyperplastic hematopoietic tissue, rupture of the tubular basement membrane, hydropic dystrophy of nephritic cells, neutrophil infiltration, fibrinoid necrosis of nephretic tubules, hemosiderin deposition, and edema. The liver sections revealed lymphocyte infiltration, dilation of hepatic sinusoids, expansion of space between hepatic sinusoids, and focal necrosis. The inflammatory responses observed in kidney and liver in the present study were presumably suppuration and were attributed to the potential virulence factors of E. tarda.

scholarly journals Antagonistic activity of terrestrial Streptomyces sp. VITNK9 against Gram negative bacterial pathogens affecting the fish and shellfish in aquaculture

2018 ◽  
Vol 53 (2) ◽  
pp. 171 ◽  
Author(s):  
Mohammed Ishaque Nabila ◽  
Krishnan Kannabiran
Keyword(s):  
Antibacterial Activity ◽  
Ethyl Acetate ◽  
Vibrio Harveyi ◽  
Bacterial Pathogens ◽  
Vibrio Anguillarum ◽  
Antagonistic Activity ◽  
Edwardsiella Tarda ◽  
Streptomyces Sp ◽  
Aeromonas Caviae ◽  
Fish And Shellfish

A total of 72 morphologically different actinomycetes isolates were isolated from samples collected at different regions of Vellore, Tamil Nadu, India and screened for its antibacterial activity against fish and shellfish pathogens. All actinomycetes isolates were screened for antibacterial activity by cross streak method against the selected fish and shellfish bacterial pathogens including Aeromonas caviae, Aeromonas hydroplila, Edwardsiella tarda, Vibrio anguillarum and Vibrio harveyi. Secondary screening of antagonistic isolates by well diffusion method leads to the identification of potential isolate. Culture conditions for the potential isolate were optimized for maximal growth and yield of the ethyl acetate (EA) crude extract. The potential isolate was characterized by molecular taxonomy and phylogeny and identified as Streptomyces species and named as Streptomyces sp. VITNK9. The 16S rDNA nucleotide sequence was searched through the GenBank database and showed 83% similarity to Streptomyces vinaceusdrappus. The EA extract prepared from Streptomyces sp. VITNK9 showed moderate antagonistic activity accessed by the formation of zone of growth inhibition against, Aeromonas caviae (15.33 mm), Aeromonas hydrophila (17.66 mm), Edwardsiella tarda (18.33 mm), Vibrio anguillarum (14.33 mm) and Vibrio harveyi (14.33 mm). The MIC value of EA extract was ranged between 0.03-0.125 mg mL-1. The GC-MS spectrum of the ethyl acetate extract revealed the presence of two major compounds, pyrrolo [1,2-A] pyrazine-1,4-Dione (56.67%) and Hexahydro-3-(2-Methylpropyl) (27.91%), respectively. The results of the study suggest that Streptomyces sp. VITNK9 is a potential source for antagonistic secondary metabolites against fish and shellfish bacterial pathogens.

scholarly journals Detection of Trypanosoma cruzi and Trypanosoma rangeli Infection by Duplex PCR Assay Based on Telomeric Sequences

2003 ◽  
Vol 10 (5) ◽  
pp. 775-779 ◽  
Author(s):  
Miguel Angel Chiurillo ◽  
Gladys Crisante ◽  
Agustina Rojas ◽  
Andreina Peralta ◽  
Manuel Dias ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Human Subjects ◽  
Simultaneous Detection ◽  
Duplex Pcr ◽  
Trypanosoma Rangeli ◽  
Pcr Assay ◽  
Telomeric Sequences ◽  
Species Specific ◽  
Duplex Pcr Assay ◽  
Reduviid Bugs

ABSTRACT We used the species specificity and repetitious nature of subtelomeric kinetoplastida sequences to generate a duplex PCR assay for the simultaneous detection of Trypanosoma cruzi and Trypanosoma rangeli in experimentally and naturally infected triatomine (Reduviid) bugs and in infected human subjects. The assay was species specific and was capable of detecting 1/20th of T. cruzi and 1/4th of T. rangeli cell equivalents without complementary hybridization. In addition, the PCR-based assay was robust enough for direct application to difficult biological samples such as Reduviid feces or guts and was capable of recognizing all T. cruzi and T. rangeli strains and lineages. Because the assay primers amplify entirely different target sequences, no reaction interference was observed, facilitating future adaptation of this assay to an automated format.

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