SN/T 2124-2013Quarantine protocol for infection with Gyrodactylus safaris (English PDF)

Foreword

This standard was drafted in accordance with GB/T 1.1-2009 given rules.

The standard reference to the World Organisation for Animal Health (OIE) of "aquatic animal disease diagnostic manual (2011 Edition)" on 2.3.03 Atlantic salmon Relevant sections of three generations of echinococcosis, combined with data reported in the literature as well as the drafting of many years of work experience in quarantine at home and abroad.

Instead of the standard SN/T 2124-2008 "Atlantic salmon Gyrodactylus quarantine technical specifications." this standard compared with SN/T 2124-2008, the main technical changes are as follows.

--- Simplifies the morphological identification methods, no longer used as the main basis for the determination of the species;

--- Remove the IGS sequences by PCR method;

--- Increase Atlantic salmon Gyrodactylus chart, mark the subsequent absorption structure, increase digestion and parasite species identification based hook sickle method.

This standard is proposed and managed by the National Certification and Accreditation Administration Committee.

This standard was drafted. Chinese Academy of Inspection and Quarantine, Shenzhen People's Republic of China Exit Inspection and Quarantine, the People's Republic of China State and Shandong Entry-Exit Inspection and Quarantine, Ningbo People's Republic of China.

1 Scope

This standard specifies the Atlantic salmon three generations of echinococcosis quarantine when sample collection, morphological examination, polymerase chain reaction (PCR) screening and species Identification of class.

This standard applies to the three generations of Atlantic salmon echinococcosis quarantine and epidemiological investigation.

2 Normative references

The following documents for the application of this document is essential. For dated references, only the dated version suitable for use herein Member. For undated references, the latest edition (including any amendments) applies to this document.

Laboratory use specifications and test methods GB/T 6682 Analysis GB/T 18088 Entry and Exit Animal Quarantine sampling

3 Abbreviations

The following abbreviations apply to this document.

ITS. amplified internal transcribed spacer (internaltranscribedspacer) COI. mitochondrial cytochrome oxidase I gene (mitochondrialcytochromeCoxidaseI)

4 Pathogen Overview

Atlantic salmon Gyrodactylus (Gyrodactylussalaris) were responsible for a three generations of Atlantic salmon viviparous freshwater parasite coccidiosis. From the morphological Point of view, and belong to the Atlantic salmon Gyrodactylus Gyrodactylus G.teuchis (mainly parasitic on brown trout, Atlantic salmon and rainbow trout) and G.thymalli (main to parasite in trout) are very similar, easily confused with each other, the specific circumstances see Appendix A.

5 Material

Unless otherwise specified, the use of chemical reagents were of analytical grade. Test water required to achieve the GB/T 6682 in a water requirements.

5.1 Ammonium bitterness - glycerol (APG) solution. preparation methods, see Appendix B, B.1.

5.2 soft tissue digestion, digestive stop solution. preparation methods see B.2, B.3.

5.3 lysate. preparation methods, see B.4.

5.4 Primer. concentration 10µmol/L, which sequence is as follows.

ITS-up. 5'-TTTCCGTAGGTGAACCT-3 ' ITS-lp. 5'-TCCTCCGCTTAGTGATA-3 ' COI-up. 5'-TAATCGGCGGGTTCGGTAA-3 ' COI-lp. 5'-GAACCATGTATCGTGTAGCA-3 '

5.5 PCR buffer, dNTP, DNA polymerase, DNAMarker, electrophoresis sample buffer and the like, are commercially kit components.

5.6 electrophoresis buffer. preparation methods see B.5.