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

Shinn, Andrew P.; Collins, Catherine; García‐Vásquez, Adriana; Snow, M. H.; Matějusová, Iveta; Paladini, Giuseppe; Longshaw, Matt; Lindenstrøm, Thomas; Stone, David M.; Turnbull, James; Picón-Camacho, Sara M.; Rivera, C. Vázquez; Duguid, R. A.; Mo, Tor Atle; Hansen, Haakon; Olstad, Kjetil; Cable, Joanne; Harris, P. D.; Kerr, Rose; Graham, Deena; Monaghan, Sean J.; Yoon, Gil Ha; Buchmann, Kurt; Taylor, Nicholas G H; Bakke, Tor A.; Raynard, R. S.; Irving, S. N.; Bron, James E.

doi:10.1016/j.ijpara.2010.04.016

Cited by 22 publications

(27 citation statements)

References 30 publications

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“…To establish the identity of the different Gyrodactylus species we used information from the first description and follow-up descriptive literature [9,48]. We also used an additional visual identification as recommended by Shinn et al [49] comparing our samples with pictures from the database GyroDb [22]. Five species of Gyrodactylus have previously been reported on minnow from Norway: G. laevis, Malmberg 1957, G. magnificus, Malmberg 1957, G. phoxini, Malmberg 1957, G. macronychus, Malmberg 1957, G. aphyae, Malmberg 1957.…”

Section: Gyrodactylus Spp Identificationmentioning

confidence: 99%

Genetic population structure ofGyrodactylus thymalli(Monogenea) in a large Norwegian river system

Pettersen

Hansen

et al. 2015

Parasitology

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The extent of geographic genetic variation is the result of several processes such as mutation, gene flow, selection and drift. Processes that structure the populations of parasite species are often directly linked to the processes that influence the host. Here, we investigate the genetic population structure of the ectoparasite Gyrodactylus thymalli Žitňan, 1960 (Monogenea) collected from grayling (Thymallus thymallus L.) throughout the river Glomma, the largest watercourse in Norway. Parts of the mitochondrial dehydrogenase subunit 5 (NADH 5) and cytochrome oxidase I (COI) genes from 309 G. thymalli were analysed to study the genetic variation and investigated the geographical distribution of parasite haplotypes. Three main clusters of haplotypes dominated the three distinct geographic parts of the river system; one cluster dominated in the western main stem of the river, one in the eastern and one in the lower part. There was a positive correlation between pairwise genetic distance and hydrographic distance. The results indicate restricted gene flow between sub-populations of G. thymalli, most likely due to barriers that limit upstream migration of infected grayling. More than 80% of the populations had private haplotypes, also indicating long-time isolation of sub-populations. According to a molecular clock calibration, much of the haplotype diversity of G. thymalli in the river Glomma has developed after the last glaciation.

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Section: Gyrodactylus Spp Identificationmentioning

confidence: 99%

Genetic population structure ofGyrodactylus thymalli(Monogenea) in a large Norwegian river system

Pettersen

Hansen

et al. 2015

Parasitology

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“…Hajibabaei et al 2006;King and Porter 2004;Nagy 2010;Post et al 1993;Quicke et al 1999;Srinivasan et al 2002) and is routinely used for DNA studies in helminths (e.g. Shinn et al 2010).…”

Section: Ethanol Vs Formalin and Dmsomentioning

confidence: 99%

A quick and simple method, usable in the field, for collecting parasites in suitable condition for both morphological and molecular studies

2012

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Many methods have been proposed for collecting and fixing parasites, but most were written before the molecular age, and were intended to be practised by experienced parasitologists in well-equipped laboratories. We describe here a very simple method, illustrated by photographs, for collecting helminths from the digestive tract of vertebrates. It only requires a few plastic vials, some ethanol and a means to heat water. Basically, the method consists of: (a) the extraction of all organs from the abdominal cavity; (b) opening the digestive system longitudinally; (c) agitate gut and contents in a saline solution (i.e. ca. 9% NaCl or 1/4 sea water in tap water); (d) decant in saline as many times as needed to clean contents; (e) immediately fix parasites in near-boiling saline; (f) discard saline and keep specimens in 95% ethanol. Additional information is given for collecting parasites from fish gills with a similar process. The method will collect most helminths (digeneans, larval cestodes, nematodes, acanthocephalans) from the digestive tract, and monogeneans and isopod and copepod crustaceans from fish gills. The specimens will be suitable for both morphological study and DNA sequencing. The method is simple, fast, inexpensive and can be used by untrained personnel, even in the field without electricity and without a binocular microscope. It can also be used by trained parasitologists who need to expedite treatment of abundant samples.

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“…thymalli G. derjavinoides, G. truttae) and non-target species (G. teuchis, G. lucii) from widespread geographic areas. These specimens had previously been characterised using morphology, statistical classifiers and ITS rDNA RFLP as part of a comparative proficiency test of established diagnostic methods (Shinn et al 2010) (Table 2). All specimens had been characterised by each method and the ITS rDNA and COI mtDNA sequenced from a number of them.…”

Section: Rationale Of Approachmentioning

confidence: 99%

Development of a real-time PCR assay for the identification of Gyrodactylus parasites infecting salmonids in northern Europe

Collins¹,

Kerr²,

McIntosh³

et al. 2010

Dis. Aquat. Org.

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Gyrodactylus salaris is a monogenean freshwater parasite that causes high mortality in wild Atlantic salmon, and a number of countries employ monitoring programmes for its presence. A TaqMan®-MGB (minor groove binding) probe real-time multiplex assay targeting the internal transcribed spacer ribosomal DNA (ITS rDNA) was developed to simultaneously identify G. salaris/ G. thymalli and 2 other commonly occurring Gyrodactylus species infecting salmonids in northern Europe: G. derjavinoides and G. truttae. In addition, a Gyrodactylus genus-level assay was developed to assess parasite DNA quality. The species-specific real-time PCR method correctly identified target species from a wide geographical range and from a number of salmonid hosts. It did not amplify G. lucii or G. teuchis. These species were successfully amplified using the Gyrodactylus genus real-time assay. The species-specific real-time assay proved to be significantly faster than the currently employed molecular screening method of ITS rDNA PCR amplification followed by restriction fragment length polymorphism analyses (RFLP). However, as with ITS RFLP, the real-time method did not distinguish between G. salaris and the non-pathogenic G. thymalli, its principle advantage being a significant reduction in time to achieve an initial diagnostic screen before the employment of more in-depth analyses for those specimens giving a positive G. salaris/G. thymalli real-time result.

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

Cited by 22 publications

References 30 publications

Genetic population structure ofGyrodactylus thymalli(Monogenea) in a large Norwegian river system

Genetic population structure ofGyrodactylus thymalli(Monogenea) in a large Norwegian river system

A quick and simple method, usable in the field, for collecting parasites in suitable condition for both morphological and molecular studies

Development of a real-time PCR assay for the identification of Gyrodactylus parasites infecting salmonids in northern Europe

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