scholarly journals Development and validation of an RNA- and DNA-based quantitative PCR assay for determination of Kudoa thyrsites infection levels in Atlantic salmon Salmo salar

2007 ◽  
Vol 75 ◽  
pp. 239-249 ◽  
Author(s):  
VA Funk ◽  
M Raap ◽  
K Sojonky ◽  
S Jones ◽  
J Robinson ◽  
...  
Keyword(s):  
Atlantic Salmon ◽  
Quantitative Pcr ◽  
Salmo Salar ◽  
Pcr Assay ◽  
Atlantic Salmon Salmo Salar ◽  
Kudoa Thyrsites ◽  
Development And Validation ◽  
Rna And Dna ◽  
Quantitative Pcr Assay

scholarly journals Development and evaluation of SNP panels for the detection of hybridization between wild and escaped Atlantic salmon (Salmo salar) in the western Atlantic

2019 ◽  
Vol 76 (5) ◽  
pp. 695-704 ◽  
Author(s):  
Brendan F. Wringe ◽  
Eric C. Anderson ◽  
Nicholas W. Jeffery ◽  
Ryan R.E. Stanley ◽  
Ian R. Bradbury
Keyword(s):  
Atlantic Salmon ◽  
Salmo Salar ◽  
Wild Populations ◽  
Western Atlantic ◽  
Farmed Salmon ◽  
Atlantic Salmon Salmo Salar ◽  
Hybrid Class ◽  
Snp Panels ◽  
The Stability ◽  
Development And Validation

Hybridization between wild and escaped cultured Atlantic salmon (Salmo salar) can threaten the stability and persistence of locally adapted wild populations. Here we describe the development and validation of a genomic-based approach to quantify recent hybridization between escapee and wild salmon in the western Atlantic. Based on genome-wide single nucleotide polymorphism (SNP) scans of wild and cultured salmon, collectively diagnostic panels were created for Newfoundland and the Canadian Maritimes. These panels were capable of both discriminating hybrids from nonhybrids and of correctly assigning individuals to hybrid class (i.e., pure wild, pure farm, F1, F2, and backcrosses) with a high degree of accuracy (Newfoundland 96 SNPs > 90%, Maritimes 720 SNPs > 80%). These genomic panels permit the assessment of the impacts of past and future farmed salmon escape events on wild populations and can inform the protection and conservation of wild Atlantic salmon genetic integrity in the western Atlantic.

scholarly journals Determination of RhD Zygosity: Comparison of a Double Amplification Refractory Mutation System Approach and a Multiplex Real-Time Quantitative PCR Approach

2001 ◽  
Vol 47 (4) ◽  
pp. 667-672 ◽  
Author(s):  
Rossa W K Chiu ◽  
Michael F Murphy ◽  
Carrie Fidler ◽  
Benny C Y Zee ◽  
James S Wainscoat ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Gene Dosage ◽  
Amplification Refractory Mutation System ◽  
Pcr Assay ◽  
Real Time Quantitative Pcr ◽  
Mutation System ◽  
Allele Specific ◽  
Quantitative Pcr Assay

Abstract Background: Rh isoimmunization and hemolytic disease of the newborn still occur despite the availability of Rh immunoglobulin. For the prenatal investigation of sensitized RhD-negative pregnant women, determination of the zygosity of the RhD-positive father has important implications. The currently available molecular methods for RhD zygosity assessment, in general, are technically demanding and labor-intensive. Therefore, at present, rhesus genotype assessment is most commonly inferred from results of serological tests. The recent elucidation of the genetic structure of the prevalent RHD deletion in Caucasians, as well as the development of real-time PCR, allowed us to explore two new approaches for the molecular determination of RhD zygosity. Methods: Two methods for RhD zygosity determination were developed. The first was based on the double Amplification Refractory Mutation System (double ARMS). The second was based on multiplex real-time quantitative PCR. For the double ARMS assay, allele-specific primers were designed to directly amplify the most prevalent RHD deletion found in RhD-negative individuals in the Caucasian population. The multiplex real-time quantitative PCR assay, on the other hand, involved coamplification and quantification of RHD-specific sequences in relation to a reference gene, albumin, in a single PCR reaction. A ratio, ΔCt, based on the threshold cycle, was then determined and reflects the RHD gene dosage. Results: The allele-specific primers of the double ARMS assay reliably amplified the RHD-deleted allele and therefore accurately distinguished homozygous from heterozygous RhD-positive samples. The results were in complete concordance with serological testing. For the multiplex real-time quantitative PCR assay, the ΔCt values clearly segregated into two distinct populations according to the RHD gene dosage, with mean values of 1.70 (SD, 0.17) and 2.62 (SD, 0.29) for the homozygous and heterozygous samples, respectively (P <0.001, t-test). The results were in complete concordance with the results of serological testing as well as with the double ARMS assay. Conclusion: Double ARMS and real-time quantitative PCR are alternative robust assays for the determination of RhD zygosity.

Determination of Sexual Maturation Stages of Atlantic Salmon (Salmo salar) Captured at Sea

1981 ◽  
Vol 38 (4) ◽  
pp. 405-413 ◽  
Author(s):  
D. R. Idler ◽  
S. J. Hwang ◽  
L. W. Crim ◽  
D. Reddin
Keyword(s):  
Atlantic Salmon ◽  
Salmo Salar ◽  
Gonadosomatic Index ◽  
Gonadal Development ◽  
Maturation Stage ◽  
Female Fish ◽  
Histological Techniques ◽  
Atlantic Salmon Salmo Salar ◽  
Placentia Bay

Radioimmunoassay (RIA) of plasma vitellogenin (Vg), estradiol (E2), 11-ketotestosterone (11-keto), and gonadotropin (GtH), together with histological techniques were evaluated for determination of the maturation stage of Atlantic salmon (Salmo salar) at sea.Male salmon had lower plasma Vg, E2, and higher 11-keto levels and could be distinguished from females several months in advance of spawning. Six female salmon were tagged at sea in 1975 in Placentia Bay and samples of blood taken. When the fish were recovered in rivers the lowest plasma Vg value was 396 μg/mL. This formed the basis of a working hypothesis, that fish with Vg values in excess of 396 μg/mL (~5–6 mo before spawning) will spawn in the year of capture. This hypothesis was supported by the Vg values of an additional 19 tagged females recaptured in Newfoundland and mainland rivers and brackish water off mainland rivers in 1976.Significant correlations between stage of gonadal development and plasma Vg, stage of gonadal development and gonadosomatic index (GSI), stage of gonadal development and plasma E2, plasma Vg and plasma E2, GSI and plasma E2, and between plasma Vg and 11-keto values were found for the female fish. The gonadal development of female fish from Bonavista Bay ranged between oil globule stage and secondary yolk stage. All those females which had reached the primary yolk stage would almost certainly mature and may be considered spawners of the year; on this basis 91% of the females were spawners of the year. Based on the minimum plasma Vg values, at sea, for fish which subsequently returned to the rivers, there were 86% spawners of the year among female salmon taken in Bonavista Bay. Based on plasma Vg levels, spawners of the year ranged from essentially zero in Greenland (fish captured in Greenland during August–November are not spawners of the year, except for the very few that spawn in one Greenland river) to 100% for fish caught in several other fisheries.11-Ketotestosterone was higher in the male fish than in the female fish and there was a correlation between GSI and 11-keto for male fish.The plasma GtH content of fish taken in the sea was extremely low as measured by RIA.Key words: salmon, maturation, migration, radioimmunoassay, vitellogenin, estradiol, gonadotropin, 11-ketotestosterone, histology, gonadosomatic index

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