TY - JOUR
T1 - Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)
AU - Shinn, A. P.
AU - Collins, C.
AU - García-Vásquez, A.
AU - Snow, M.
AU - Matějusová, I.
AU - Paladini, G.
AU - Longshaw, M.
AU - Lindenstrøm, T.
AU - Stone, D. M.
AU - Turnbull, J. F.
AU - Picon-Camacho, S. M.
AU - Rivera, C. Vázquez
AU - Duguid, R. A.
AU - Mo, T. A.
AU - Hansen, H.
AU - Olstad, K.
AU - Cable, J.
AU - Harris, P. D.
AU - Kerr, R.
AU - Graham, D.
AU - Monaghan, S. J.
AU - Yoon, G. H.
AU - Buchmann, K.
AU - Taylor, N. G.H.
AU - Bakke, T. A.
AU - Raynard, R.
AU - Irving, S.
AU - Bron, J. E.
N1 - Funding Information:
The authors gratefully acknowledge the financial support provided by the diagnostic services of both Cefas Weymouth Laboratory , UK and the Marine Laboratory, Marine Science Scotland , Aberdeen. The authors also thank the staff of the freshwater fisheries services operating in each country notably Kevin Denham and Richard Gardiner (Cefas Weymouth Laboratory), Helen Rowley and Amanda Brown (Disease Surveillance & Investigation Dept., Veterinary Sciences Division, Agri-food & Biosciences Institute, Belfast, Northern Ireland) for the provision of Gyrodactylus infected salmonid and pike material for study.
PY - 2010/10
Y1 - 2010/10
N2 - Despite routine screening requirements for the notifiable fish pathogen Gyrodactylus salaris, no standard operating procedure exists for its rapid identification and discrimination from other species of Gyrodactylus. This study assessed screening and identification efficiencies under real-world conditions for the most commonly employed identification methodologies: visual, morphometric and molecular analyses. Obtained data were used to design a best-practice processing and decision-making protocol allowing rapid specimen throughput and maximal classification accuracy. True specimen identities were established using a consensus from all three identification methods, coupled with the use of host and location information. The most experienced salmonid gyrodactylid expert correctly identified 95.1% of G. salaris specimens. Statistical methods of classification identified 66.7% of the G. salaris, demonstrating the need for much wider training. Molecular techniques (internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP)/cytochrome c oxidase I (COI) sequencing) conducted in the diagnostic laboratory most experienced in the analysis of gyrodactylid material, identified 100% of the true G. salaris specimens. Taking into account causes of potential specimen loss, the probabilities of a specimen being accurately identified were 95%, 87% and 92% for visual, morphometric and molecular techniques, respectively, and the probabilities of correctly identifying a specimen of G. salaris by each method were 81%, 58% and 92%. Inter-analyst agreement for 189 gyrodactylids assessed by all three methods using Fleiss' Kappa suggested substantial agreement in identification between the methods. During routine surveillance periods when low numbers of specimens are analysed, we recommend that specimens be analysed using the ITS-RFLP approach followed by sequencing of specimens with a "G. salaris-like" (i.e. G. salaris, Gyrodactylus thymalli) banding pattern. During periods of suspected outbreaks, where a high volume of specimens is expected, we recommended that specimens be identified using visual identification, as the fastest processing method, to select "G. salaris-like" specimens, which are subsequently identified by molecular-based techniques.
AB - Despite routine screening requirements for the notifiable fish pathogen Gyrodactylus salaris, no standard operating procedure exists for its rapid identification and discrimination from other species of Gyrodactylus. This study assessed screening and identification efficiencies under real-world conditions for the most commonly employed identification methodologies: visual, morphometric and molecular analyses. Obtained data were used to design a best-practice processing and decision-making protocol allowing rapid specimen throughput and maximal classification accuracy. True specimen identities were established using a consensus from all three identification methods, coupled with the use of host and location information. The most experienced salmonid gyrodactylid expert correctly identified 95.1% of G. salaris specimens. Statistical methods of classification identified 66.7% of the G. salaris, demonstrating the need for much wider training. Molecular techniques (internal transcribed spacer region-restriction fragment length polymorphism (ITS-RFLP)/cytochrome c oxidase I (COI) sequencing) conducted in the diagnostic laboratory most experienced in the analysis of gyrodactylid material, identified 100% of the true G. salaris specimens. Taking into account causes of potential specimen loss, the probabilities of a specimen being accurately identified were 95%, 87% and 92% for visual, morphometric and molecular techniques, respectively, and the probabilities of correctly identifying a specimen of G. salaris by each method were 81%, 58% and 92%. Inter-analyst agreement for 189 gyrodactylids assessed by all three methods using Fleiss' Kappa suggested substantial agreement in identification between the methods. During routine surveillance periods when low numbers of specimens are analysed, we recommend that specimens be analysed using the ITS-RFLP approach followed by sequencing of specimens with a "G. salaris-like" (i.e. G. salaris, Gyrodactylus thymalli) banding pattern. During periods of suspected outbreaks, where a high volume of specimens is expected, we recommended that specimens be identified using visual identification, as the fastest processing method, to select "G. salaris-like" specimens, which are subsequently identified by molecular-based techniques.
KW - Contingency planning
KW - Gyrodactylus salaris
KW - Identification
KW - Monogenea
KW - Pathogen introduction
KW - Protocol
KW - Validation
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U2 - 10.1016/j.ijpara.2010.04.016
DO - 10.1016/j.ijpara.2010.04.016
M3 - Article
C2 - 20595003
AN - SCOPUS:82155194100
SN - 0020-7519
VL - 40
SP - 1455
EP - 1467
JO - International Journal for Parasitology
JF - International Journal for Parasitology
IS - 12
ER -