Hostname: page-component-8448b6f56d-m8qmq Total loading time: 0 Render date: 2024-04-24T05:03:27.924Z Has data issue: false hasContentIssue false
  • English
  • Français

Genotyping of individual Ceratonova shasta (Cnidaria: Myxosporea) myxospores reveals intra-spore ITS-1 variation and invalidates the distinction of genotypes II and III

Published online by Cambridge University Press:  27 March 2018

Stephen D. Atkinson ,
Sascha L. Hallett  and
Jerri L. Bartholomew
Stephen D. Atkinson*
Affiliation:
Department of Microbiology, Oregon State University, 226 Nash Hall, Corvallis, Oregon 97331, USA
Sascha L. Hallett
Affiliation:
Department of Microbiology, Oregon State University, 226 Nash Hall, Corvallis, Oregon 97331, USA
Jerri L. Bartholomew
Affiliation:
Department of Microbiology, Oregon State University, 226 Nash Hall, Corvallis, Oregon 97331, USA
*
Author for correspondence: Stephen Atkinson, E-mail: atkinsos@oregonstate.edu
Get access Rights & Permissions [Opens in a new window]

Abstract

Genotypes of the myxosporean parasite Ceratonova shasta are defined by the number of ATC repeats in the parasite's ribosomal DNA internal transcribed spacer region 1. These genotypes correlate with specific salmonid fish hosts. We observed coho salmon (Oncorhynchus kisutch) and rainbow trout (Oncorhynchus mykiss) with mixtures of genotypes II and III, and assumed that this was a consequence of fish having an aggregate infection from multiple individual parasites. We hypothesized that although multiple ITS copies are present within a parasite spore, the DNA sequences of these copies are identical, and thus individual C. shasta spores are a single genotype. We tested this by extracting and sequencing DNA from individual myxospores. We trialed three approaches for in-tube DNA extraction; digestion with proteinase K was superior to simply rehydrating spores, or incubation in the buffer. Sequences from 14 myxospores were each a mixture of genotypes II and III. Therefore, intra-genomic ribosomal DNA variants exist within individual parasite spores, and II and III should no longer be regarded as discrete C. shasta genotypes. This single-spore genotyping approach will be a useful tool for testing validity of other C. shasta genotypes, and for correctly matching genotype with phenotype for mixed infections of other myxozoan species.

Keywords

Ceratonova shastaITS-1 genotypemyxozoanwithin-spore genotypes
Type
Research Article
Information
Parasitology , Volume 145 , Issue 12 , October 2018 , pp. 1588 - 1593
Check for updates
Copyright
Copyright © Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

BiOp (2013) Biological Opinions on the Effects of Proposed Klamath Project Operations from May 31, 2013, through March 31, 2023, on Five Federally Listed Threatened and Endangered Species. Prepared By: National Marine Fisheries Service Southwest Region Northern California Office and U.S. Fish and Wildlife Service Pacific Southwest Region Klamath Falls Fish and Wildlife Office. NMFS file number: SWR-2012-9372, FWS file number: 08EKLA00-2013-F-0014.Google Scholar
Alexander, JD, Hallett, SL, Stocking, RW, Xue, L and Bartholomew, JL (2014) Host and parasite populations after a ten year flood: Manayunkia speciosa and Ceratonova shasta in the Klamath River. Northwest Science 88, 219–233.Google Scholar
Atkinson, SD and Bartholomew, JL (2010 a) Disparate infection patterns of Ceratomyxa shasta (Myxozoa) in rainbow trout Oncorhynchus mykiss and Chinook salmon Oncorhynchus tshawytscha correlate with ITS-1 sequence variation in the parasite. International Journal for Parasitology 40, 599–604.Google Scholar
Atkinson, SD and Bartholomew, JL (2010 b) Spatial, temporal and host factors structure the Ceratomyxa shasta (Myxozoa) population in the Klamath River Basin. Infection, Genetics and Evolution 10, 1019–1026.Google Scholar
Atkinson, SD, Foott, JS and Bartholomew, JL (2014) Erection of Ceratonova n. gen. (Myxosporea:Ceratomyxidae) to encompass freshwater species C. gasterostea n. sp. from threespine stickleback (Gasterosteus aculeatus) and C. shasta n. comb. from salmonid fishes. Journal of Parasitology 100, 640–645.Google Scholar
Bart, A, van der Heijden, HM, Greve, S, Speijer, D, Landman, WJ and van Gool, T (2008) Intragenomic variation in the internal transcribed spacer 1 region of Dientamoeba fragilis as a molecular epidemiological marker. Journal of Clinical Microbiology 46, 3270–3275.Google Scholar
Bartholomew, JL (1998) Host resistance to infection by the myxosporean parasite Ceratomyxa shasta: a review. Journal of Aquatic Animal Health 10, 112–120.Google Scholar
Bartholomew, JL, Whipple, MJ, Stevens, DG and Fryer, JL (1997) The life cycle of Ceratomyxa shasta, a myxosporean parasite of salmonids, requires a freshwater polychaete as an alternate host. Journal of Parasitology 83, 859–868.Google Scholar
Bartošová, P, Fiala, I, Jirku, M, Cinková, M, Caffara, M, Fioravanti, ML, Atkinson, SD, Bartholomew, JL and Holzer, AS (2013) Sphaerospora sensu stricto: taxonomy,diversity and evolution of a unique lineage of myxosporeans (Myxozoa). Molecular Phylogenetics and Evolution 68, 93–105.Google Scholar
Bjork, SJ and Bartholomew, JL (2009) The effects of water velocity on the Ceratomyxa shasta infectious cycle. Journal of Fish Diseases 32, 131–142.Google Scholar
El-Matbouli, M and Hoffmann, RW (1998) Light and electron microscopic studies on the chronological development of Myxobolus cerebralis to the actinosporean stage in Tubifex tubifex. International Journal of Parasitology 28, 195–217.Google Scholar
Eszterbauer, E (2004) Genetic relationship among gill-infecting Myxobolus species (Myxosporea) of cyprinids: molecular evidence of importance of tissue-specificity. Diseases of Aquatic Organisms 58, 35–40.Google Scholar
Feist, S, Morris, DJ, Alama-Bermejo, G and Holzer, AS (2015) Cellular processes in myxozoans. In Okamura, B, Gruhl, A and Bartholomew, JL (eds). Myxozoan Evolution, Ecology and Development. Switzerland: Springer International Publishing, pp. 139–154.Google Scholar
Fiala, I (2006) The phylogeny of Myxosporea (Myxozoa) based on small subunit ribosomal RNA gene analysis. International Journal for Parasitology 36, 1521–1534.Google Scholar
Gong, J, Dong, J, Xihan, L and Massana, R (2013) Extremely high copy numbers and polymorphisms of the rDNA operon estimated from single cell analysis of oligotrich and peritrich ciliates. Protist 164, 369–379.Google Scholar
Hall, TA (1999) Bioedit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41, 95–98.Google Scholar
Hallett, SL and Bartholomew, JL (2006) Application of a real-time PCR assay to detect and quantify the myxozoan parasite Ceratomyxa shasta in river water samples. Diseases of Aquatic Organisms 71, 109–118.Google Scholar
Hallett, SL and Diamant, A (2001) Ultrastructure and small-subunit ribosomal DNA sequence of Henneguya lesteri n.sp. (Myxosporea), a parasite of sand whiting Sillago analis (Sillaginidae) from the coast of Queensland, Australia. Diseases of Aquatic Organisms 46, 197–212.Google Scholar
Hallett, SL, Ray, RA, Hurst, CN, Holt, RA, Buckles, GR, Atkinson, SD and Bartholomew, JL (2012) Density of the waterborne parasite, Ceratomyxa shasta, and its biological effects on salmon. Applied and Environmental Microbiology 78, 3724–3731.Google Scholar
Heiniger, H and Adlard, RD (2014) Relatedness of novel species of Myxidium Buetschli, 1882, Zschokkella Auerbach, 1910 and Ellipsomyxa Køie, 2003 (Myxosporea: Bivalvulida) from the gall bladders of marine fishes (Teleostei) from Australian waters. Systematic Parasitology 87, 47–72.Google Scholar
Henderson, M and Okamura, B (2004) The phylogeography of salmonid proliferative kidney disease in Europe and North America. Proceedings of the Royal Society London B Biological Science 271, 1729–1736.Google Scholar
Hendrickson, GL, Carleton, A and Manzer, D (1989) Geographic and seasonal distribution of the infective stage of Ceratomyxa shasta (Myxozoa) in Northern California. Diseases of Aquatic Organisms 7, 165–169.Google Scholar
Hillis, DM and Davis, SK (1988) Ribosomal DNA: intraspecific polymorphism, concerted evolution, and phylogeny reconstruction. Systematic Zoology 37, 63–66.Google Scholar
Hurst, CN and Bartholomew, JL (2012) Ceratomyxa shasta genotypes cause differential mortality in their salmonids hosts. Journal of Fish Diseases 35, 725–732.Google Scholar
Hurst, CN, Wong, P, Hallett, SL, Ray, RA and Bartholomew, JL (2014) Transmission and persistence of Ceratonova shasta genotypes in Chinook Salmon. Parasitology 100, 773–777.Google Scholar
Keller, I, Chintauan-Marquier, IC, Veltsos, P and Nichols, RA (2006) Ribosomal DNA in the grasshopper Podisma pedestris: escape from concerted evolution. Genetics 174, 863–874.Google Scholar
Liu, Y, Whipps, CM, Nie, P and Gu, Z (2014) Myxobolus oralis sp. n. (Myxosporea: Bivalvulida) infecting the palate in the mouth of gibel carp Carassius auratus gibelio (Cypriniformes: Cyprinidae). Folia Parasitologica 61, 505–511.Google Scholar
Meaders, MD and Hendrickson, GL (2009) Chronological development of Ceratomyxa shasta in the polychaete host, Manayunkia speciosa. Journal of Parasitology 95, 1397–1407.Google Scholar
Morris, DJ (2012) A new model for myxosporean (Myxozoa) development explains the endogenous budding phenomenon, the nature of cell within cell life stages and evolution of parasitism from a cnidarian ancestor. International Journal for Parasitology 42, 829–840.Google Scholar
Okamura, B, Gruhl, A and Bartholomew, JL (Eds). (2015) Myxozoan Evolution, Ecology and Development, 1st edn. Switzerland: Springer International Publishing.Google Scholar
Palenzuela, O, Tronbridge, G and Bartholomew, JL (1999) Development of a polymerase chain reaction diagnostic assay for Ceratomyxa shasta, a myxosporean parasite of salmonid fish. Diseases of Aquatic Organisms 36, 45–51.Google Scholar
Rooney, AP (2004) Mechanisms underlying the evolution and maintenance of functionally heterogeneous 18S rRNA genes in Apicomplexans. Molecular Biology and Evolution 21, 1704–1711.Google Scholar
Stinson, MET and Bartholomew, JL (2012) Predicted redistribution of Ceratomyxa shasta genotypes with salmonid passage in the Deschutes River, Oregon. Journal of Aquatic Animal Health 24, 274–280.Google Scholar
Stocking, RW and Bartholomew, JL (2007) Distribution and habitat characteristics of Manayunkia speciosa and infection prevalence with the parasite Ceratomyxa shasta in the Klamath River, Oregon-California. Journal of Parasitology 93, 78–88.Google Scholar
Whipps, CM and Kent, ML (2006) Phylogeography of the Cosmopolitan Marine Parasite Kudoa thyrsites (Myxozoa: Myxosporea). Journal of Eukaryotic Microbiology 53, 364–373.Google Scholar
Whipps, CM, El-Matbouli, M, Hedrick, RP, Blazer, V and Kent, ML (2004) Myxobolus cerebralis internal transcribed spacer 1 (ITS-1) sequences support recent spread of the parasite to North America and within Europe. Diseases of Aquatic Organisms 60, 105–108.Google Scholar
Zhai, Y-H, Zhou, L and Gui, J-F (2012) Identification and characterization of one novel type of Triactinomyxon with short spore axis. Parasitology Research 110, 2385–2393.Google Scholar