Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)

A.P. Shinn; C. Collins; A. García-Vásquez; M. Snow; I. Matejusová; G. Paladini; M. Longshaw; T. Lindenstrøm; D.M. Stone; J.F. Turnbull; S.M. Picon-Camacho; C. Vázquez Rivera; R.A. Duguid; T.A. Mo; H. Hansen; K. Olstad; J. Cable; P.D. Harris; R. Kerr; D. Graham; S.J. Monaghan; G.H. Yoon; Kurt Buchmann; N.G.H. Taylor; T.A. Bakke; R. Raynard; S. Irving; J.E. Bron

doi:10.1016/j.ijpara.2010.04.016

Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)

A.P. Shinn, C. Collins, A. García-Vásquez, M. Snow, I. Matejusová, G. Paladini, M. Longshaw, T. Lindenstrøm, D.M. Stone, J.F. Turnbull, S.M. Picon-Camacho, C. Vázquez Rivera, R.A. Duguid, T.A. Mo, H. Hansen, K. Olstad, J. Cable, P.D. Harris, R. Kerr, D. GrahamS.J. Monaghan, G.H. Yoon, Kurt Buchmann, N.G.H. Taylor, T.A. Bakke, R. Raynard, S. Irving, J.E. Bron

18 Citations (Scopus)

Abstract

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.

Original language	English
Journal	International Journal for Parasitology
Volume	40
Issue number	12
Pages (from-to)	1455-1467
Number of pages	13
ISSN	0020-7519
DOIs	https://doi.org/10.1016/j.ijpara.2010.04.016
Publication status	Published - Oct 2010

Keywords

Former LIFE faculty

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10.1016/j.ijpara.2010.04.016

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Shinn, A. P., Collins, C., García-Vásquez, A., Snow, M., Matejusová, I., Paladini, G., Longshaw, M., Lindenstrøm, T., Stone, D. M., Turnbull, J. F., Picon-Camacho, S. M., Rivera, C. V., Duguid, R. A., Mo, T. A., Hansen, H., Olstad, K., Cable, J., Harris, P. D., Kerr, R., ... Bron, J. E. (2010). Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea). International Journal for Parasitology, 40(12), 1455-1467. https://doi.org/10.1016/j.ijpara.2010.04.016

Shinn, AP, Collins, C, García-Vásquez, A, Snow, M, Matejusová, I, Paladini, G, Longshaw, M, Lindenstrøm, T, Stone, DM, Turnbull, JF, Picon-Camacho, SM, Rivera, CV, Duguid, RA, Mo, TA, Hansen, H, Olstad, K, Cable, J, Harris, PD, Kerr, R, Graham, D, Monaghan, SJ, Yoon, GH, Buchmann, K, Taylor, NGH, Bakke, TA, Raynard, R, Irving, S & Bron, JE 2010, 'Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)', International Journal for Parasitology, vol. 40, no. 12, pp. 1455-1467. https://doi.org/10.1016/j.ijpara.2010.04.016

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title = "Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)",

abstract = "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.",

keywords = "Former LIFE faculty, Gyrodactylus salaris; Contingency planning; Pathogen introduction; Validation; Identification; Monogenea; Protocol",

author = "A.P. Shinn and C. Collins and A. Garc{\'i}a-V{\'a}squez and M. Snow and I. Matejusov{\'a} and G. Paladini and M. Longshaw and T. Lindenstr{\o}m and D.M. Stone and J.F. Turnbull and S.M. Picon-Camacho and Rivera, {C. V{\'a}zquez} and R.A. Duguid and T.A. Mo and H. Hansen and K. Olstad and J. Cable and P.D. Harris and R. Kerr and D. Graham and S.J. Monaghan and G.H. Yoon and Kurt Buchmann and N.G.H. Taylor and T.A. Bakke and R. Raynard and S. Irving and J.E. Bron",

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language = "English",

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pages = "1455--1467",

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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 - Matejusová, 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, Kurt

AU - Taylor, N.G.H.

AU - Bakke, T.A.

AU - Raynard, R.

AU - Irving, S.

AU - Bron, J.E.

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 - Former LIFE faculty

KW - Gyrodactylus salaris; Contingency planning; Pathogen introduction; Validation; Identification; Monogenea; Protocol

U2 - 10.1016/j.ijpara.2010.04.016

DO - 10.1016/j.ijpara.2010.04.016

M3 - Journal article

C2 - 20595003

SN - 0020-7519

VL - 40

SP - 1455

EP - 1467

JO - International Journal for Parasitology

JF - International Journal for Parasitology

IS - 12

ER -

Multi-centre testing and validation of current protocols for the identification of Gyrodactylus salaris (Monogenea)

Abstract

Keywords

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