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New data on flatfish scuticociliatosis reveal that Miamiensis avidus and Philasterides dicentrarchi are different species

  • ANA-PAULA DE FELIPE (a1), JESÚS LAMAS (a2), ROSA-ANA SUEIRO (a1) (a2), IRIA FOLGUEIRA (a1) and JOSÉ-MANUEL LEIRO (a1)...
Summary
SUMMARY

Scuticociliatosis is a severe disease in farmed flatfish. However, the causative agent is not always accurately identified. In this study, we identified two isolates of scuticociliates from an outbreak in cultured fine flounder Paralichthys adspersus. Scuticociliate identification was based on morphological data, examination of life stages and the use of molecular approaches. The isolates were compared with a strain of Philasterides dicentrachi from turbot Scophthalmus maximus and with a strain deposited in the American Type Culture Collection as Miamiensis avidus ATCC® 50180. The use of morphological, biological and molecular methods enabled us to identify the isolates from the fine flouder as P. dicentrarchi. Comparison of P. dicentrachi isolates and M. avidus revealed some differences in the buccal apparatus. Unlike P. dicentrarchi, M. avidus has a life cycle with three forms: macrostomes (capable of feeding on P. dicentrarchi), microstomes and tomites. Additionally, we found differences in the 18S rRNA and α- and β-tubulin gene sequences, indicating that P. dicentrarchi and M. avidus are different species. We therefore reject the synonymy/conspecificity of the two taxa previously suggested. Finally, we suggest that a combination of morphological, biological, molecular (by multigene analysis) and serological techniques could improve the identification of scuticociliates parasites in fish.

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*Corresponding author: Laboratorio de Parasitología, Instituto de Investigación y Análisis Alimentarios, c/ Constantino Candeira s/n, 15782, Santiago de Compostela (A Coruña), Spain. E-mail: josemanuel.leiro@usc.es
References
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Aescht E. (2001). Generic names of ciliates (Protozoa, Ciliophora). Denisia 1, 1350.
Azad I. S., Al-Marzouk A., James C. M., Almatar S. and Al-Gharabally H. (2007). Scuticociliatosis-associated mortalities and histopathology of natural infection in cultured silver pomfret (Pampus argenteus Euphrasen) in Kuwait. Aquaculture 262, 202210.
Bonar C. J., Garner M. M., Weber E. S., Keller C. J., Murray M., Adams L. M. A. and Frasca S. Jr. (2013). Pathologic findings in weedy (Phyllopteryx taeniolatus) and leafy (Phycodurus eques) seadragons. Veterinary Pathology 50, 368376.
Bordier C. (1981). Phase separation of integral membrane proteins in Triton X-114. The Journal of Biological Chemistry 256, 16041607.
Budiño B., Lamas J., Pata M. P., Arranz J. A., Sanmartín M. L. and Leiro J. (2011 a). Intraspecific variability in several isolates of Philasterides dicentrarchi (syn. Miamiensis avidus), a scuticociliate parasite of farmed turbot. Veterinary Parasitology 175, 260272.
Budiño B., Lamas J., González A., Pata M. P., Devesa S., Arranz J. A. and Leiro J. (2011 b). Coexistence of several Philasterides dicentrarchi strains on a turbot fish farm. Aquaculture 322–323, 2332.
Budiño B., Leiro J., Cabaleiro S. and Lamas J. (2012). Characterization of Philasterides dicentrarchi isolates that are pathogenic to turbot: serology and cross-protective immunity. Aquaculture, 364–365, 130136.
Declercq A. M., Chiers K., Van den Broeck W., Rekecki A., Teerlinck S., Adriaens D., Haesebrouck F. and Decostere A. (2014). White necrotic tail tips in estuary seahorses, Hippocampus kuda, Bleeker. Journal of Fish Diseases 37, 501504.
de Waal T. (2012). Advances in diagnosis of protozoan diseases. Veterinary Parasitology 189, 6574.
di Cicco E., Paradis E., Stephen C., Turba M. E. and Rossi G. (2013). Scuticociliatid ciliate outbreak in Australian potbellied sea horse, Hippocampus abdominalis (Lesson, 1827): clinical signs, histopathological findings, and treatment with metronidazole. Journal of Zoo and Wildlife Medicine 44, 435440.
Dickerson H. W., Clark T. G. and Findly R. C. (1989). Icththyophthirius multifiliis has membrane-associated immobilization antigens. Journal of Protozoology 36, 159164.
Dragesco A., Dragesco J., Coste F., Gasc C., Romestand B., Raymond J. and Bouix G. (1995). Philasterides dicentrarchi, n. Sp. (Ciliophora, Scuticociliatida), a histiophagous opportunistic parasite of Dicentrarchus labrax (Linnaeus, 1758), a reared marine fish. European Journal of Protistology 31, 327340.
Fan X., Hu X., Al-Farraj S. A., Clamp J. C. and Song W. (2011). Morphological description of three marine ciliates (Ciliophora, Scuticociliatia), with establishment of a new genus and two species. European Journal of Protistology 47, 186196.
Felsenstein J. (1985). Confidence limits on phylogenies: and approach using the boostrap. Evolution 39, 783791.
Fenchel T. (1987). Adaptative significance of polymorphic life cycles in protozoa: resposes to starvation and reffeding in two species of marine ciliates. Journal of Experimental Marine Biology and Ecology 136, 159177.
Fernández-Galiano D. (1994). The ammoniacal silver carbonate method as a general procedure in the study of protozoa from sewage (and other) waters. Water Research 28, 495496.
Gao F., Fan X., Yi Z., Strüder-Kypke M. and Song W. (2010). Phylogenetic consideration of two scuticociliate genera, Philasterides and Boveria (Protozoa, Ciliophora) based on 18 S rRNA gene sequences. Parasitology International 59, 549555.
Gao F., Katz L. A. and Song W. (2012). Insights into the phylogenetic and taxonomy of philasterid ciliates (Protozoa, Ciliophora, Scuticociliatia) based on analyses of multiple molecular markers. Molecular Phylogenetics and Evolution 64, 308317.
Gao F., Katz L. A. and Song W. (2013). Multigene-based analyses on evolutionary phylogeny of two controversial ciliate orders: Pleuronematida and Loxocephalida (Protista, Ciliophora, Oligohymenophorea). Molecular Phylogenetics and Evolution 68, 5563.
Gómez-Saladín E. and Small E. B. (1993 a). Oral morphogenesis of the microstome to macrostome transformation in Miamiensis avidus strain Ma/2. Journal of Eukaryotic Microbiology 40, 363370.
Gómez-Saladin E. and Small E. B. (1993 b). Starvation induces tomitogenesis in Miamiensis avidus strain Ma/2. Journal of Eukaryotic Microbiology 40, 727730.
Grolière C. A. (1980). Morphologie et stomatogenèse chez deux ciliés Scuticociliatida des genres Phïlasterides Kahl, 1926 et Cyclidium O. F. Müller; 1786. Acta Protozoologica 19, 195206.
Harikrishnan R., Balasundaram C. and Heo M. S. (2010). Scuticociliatosis and its recent prophylactic measures in aquaculture with special reference to South Korea Taxonomy, diversity and diagnosis of scuticociliatosis: Part I Control strategies of scuticociliatosis: Part II. Fish and Shellfish Immunology 29, 1531.
Harikrishnan R., Jin C. N., Kim J. S., Balasundaram C. and Heo M. S. (2012). Philasterides dicentrarchi, a histophagous ciliate causing scuticociliatosis in olive flounder, Philasterides dicentrarchi – Histopathology investigations. Experimental Parasitology 130, 239245.
Hu X., Song W. and Warren A. (2009). Scuticociliatids. In Free-living Ciliates in Bohai and Yellow Sea, China (ed. Song W., Warren A. and Hu X.). Science Press, Beijing. v pp. 172–179.
Iglesias R., Paramá A., Álvarez M. F., Leiro J., Fernández J. and Sanmartín M. L. (2001). Philasterides dicentrarchi (Ciliophora, Scuticociliatida) as the causative agent of scuticociliatosis in farmed turbot, Scophthalmus maximus in Galicia (NW Spain). Diseases of Aquatic Organisms 46, 4755.
Iglesias R., Paramá A., Álvarez M. F., Leiro J., Aja C. and Sanmartín M. L. (2003 a). In vitro growth requeriments for the fish pathogen Philasterides dicentrarchi (Ciliophora, Scuticociliatida). Veterinary Parasitology 111, 1930.
Iglesias R., Paramá A., Álvarez M. F., Leiro J., Ubeira F. M. and Sanmartín M. L. (2003 b). Philasterides dicentrarchi (Ciliophora: Scuticociliatida) express surface immobilization antigens that probably induce protective immune responses in turbot. Parasitology 126, 125134.
Ito H., Fukuda Y., Murata K. and Kimura A. (1983). Transformation of intact yeast cells treated with alkali cations. Journal of Bacteriology 153, 163168.
Jung S. J., Kitamura S. I., Song J. Y., Joung I. Y. and Oh M. J. (2005). Complete small subunit rRNA gene sequence of the scuticociliate Miamiensis avidus pathogenic to olive flounder Paralichthys olivaceus . Diseases of Aquatic Organisms 64, 159162.
Jung S. J., Kitamura S. I., Song J. Y. and Oh M. J. (2007). Miamiensis avidus (Ciliophora: Scuticociliatida) causes systemic infection of olive flounder Paralichthys olivaceus and is a senior synonym of Philasterides dicentrarchi . Diseases of Aquatic Organisms 73, 227234.
Jung S. J., Im E. Y., Struder-Kypke M. C., Kitamura S. and Woo P. T. (2011). Small subunit ribosomal RNA and mitochondrial cytochrome c oxidase subunit 1 gene sequences of 21 strains of the parasitic scuticociliate Miamiensis avidus (Ciliophora, Scuticociliatia). Parasitology Research 108, 11531161.
Kaneshiro E. S., Dunham P. B. and Holz G. G. (1969). Osmorregulation in a marine ciliate, Miamiensis avidus. I. Regulation of inorganic ions and water. Biological Bulletin 136, 6575.
Kim S. M., Cho J. B., Kim S. K., Nam Y. K. and Kim K. H. (2004). Occurrence of scuticociliatosis in olive flounder Paralichthys olivaceus by Philasterides dicentrarchi (Ciliophora: scuticociliatia). Diseases of Aquatic Organisms 62, 23332338.
Kimura M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution 16, 111120.
Kumar S., Stecher G. and Tamura K. (2016). MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 18701873.
Leiro J., Siso M. I., Paramá A., Ubeira F. M. and Sanmartín M. L. (2000). RFLP analysis of PCR-amplified small subunit ribosomal DNA of three fish microsporidian species. Parasitology 124, 145151.
Long H. and Zufall R. A. (2010). Diverse modes of reproduction in the marine free-living ciliate Glauconema trihymene . BioMed Central Microbiology 10, 108.
López-López O., Fuciños P., Pastrana L., Rúa M. L., Cerdán M. E. and González-Siso M. I. (2010). Heterologous expression of an esterase from Thermus thermophilus HB27 in Saccharomyces cerevisiae . Journal of Biotechnology 145, 226232.
Lynn D. H. and Strüder-Kypke M. (2005). Scuticociliate endosymbionts of echinoids (phylum Echinodermata): phylogenetic relationships among species in the genera Entodiscus, Plagiopyliella, Thyrophylax, and Entorhipidium (phylum Ciliophora). Journal of Parasitology 91, 11901199.
Mallo N., Lamas J. and Leiro J. M. (2013). Evidence of an alternative oxidase pathway for mitochondrial respiration in the scuticociliate Philasterides dicentrarchi . Protist 164, 824836.
McWilliam H., Li W., Uludag M., Squizzato S., Park Y. M., Buso N., Cowley A. P. and López R. (2013). Analysis tool web services from the EMBL-EBI. Nucleic Acids Research 41(Web Server issue), W597W600.
Miao M., Warren A., Song W., Wang S., Shang H. and Chen Z. (2008). Analysis of internal transcribed spacer 2 (ITS2) region of scuticociliates and related taxa (Ciliophora, Oligohymenophorea) to infer their evolution and phylogeny. Protist 159, 519533.
Miao M., Wang Y., Song W., Clamp J. C. and Al-Rasheid K. A. S. (2010). Description of Eurystomatella sinica n. gen., n. sp., with establishment of a new family Eurystomatellidae n. fam. (Protista, Ciliophora, Scuticociliatia) and analyses of its phylogeny inferred from sequences of the small-subunit rRNA gene. International Journal of Systematic and Evolutionary Microbiology 60, 460468.
Moewus L. (1963). Studies on a marine parasitic ciliate as a potential virus vector. In Symp. on Marine Microbiology (ed. Oppenheimer C. H.), pp. 366379. Charles C. Thomas, Springfield, IL.
Morais P., Lamas J., Sanmartín M. L., Orallo F. and Leiro J. (2009). Resveratrol induces mitochondrial alterations, autophagy and a cryptobiosis-like state in scuticociliates. Protist 160, 552564.
Moustafa E. M. M., Naota M., Morita T., Tange N. and Shimada A. (2010). Pathological study on the scuticociliatosis affecting farmed Japanese flounder (Paralichthys olivaceus) in Japan. Journal of Veterinary Medical Science 72, 13591362.
Munday B. L., O’Donoghue P. J., Watts M., Rough K. and Hawkesford T. (1997). Fatal encephalitis due to the scuticociliate Uronema nigricans in sea-caged, southern bluefin tuna Thunnus maccoyii . Diseases of Aquatic Organisms 30, 1725.
Myburg H., Gryzenhout M., Wingfield B. D., Wingfield M. D. (2003). Conspecificity of Enothia eugeniae and Cryphonectria cubensis: a re-evaluation based on morphology and DNA sequence data. Mycoscience 44, 187196.
Ofelio C., Blanco A., Roura A., Pintado J., Pascual S. and Planas M. (2014). Isolation and molecular identification of the scuticociliate Porpostoma notata Moebius, 1888 from moribund reared Hippocampus hippocampus (L.) seahorses, by amplification of the SSU rRNA gene sequences. Journal of Fish Diseases 37, 10611065.
Pan X., Zhu M., Ma H., Al-Rasheid K. A. and Hu X. (2013). Morphology and small-subunit rRNA gene sequences of two novel marine ciliates, Metanophrys orientalis spec. nov. and Uronemella sinensis spec. nov. (Protista, Ciliophora, Scuticociliatia), with an improved diagnosis of the genus Uronemella . International Journal of Systematic and Evolutionary Microbiology 63, 3515–23.
Paramá A., Iglesias R., Álvarez M. F., Leiro J., Aja C. and Sanmartín M. L. (2003). Philasterides dicentrarchi (Ciliophora, Scuticociliatida): experimental infection and posible routes of entry in farmed turbot (Scophthalmus maximus). Aquaculture 217, 7380.
Paramá A., Arranz J. A., Álvarez M. F., Sanmartín M. L. and Leiro J. (2006). Ultrastructure and phylogeny of Philasterides dicentrarchi (Ciliophora: Scuticociliatia) from farmed turbot in NW Spain. Parasitology 132, 555564.
Piazzon C., Lamas J., Castro R., Budiño B., Cabaleiro S., Sanmartín M. L. and Leiro J. (2008). Antigenic and cross-protection studies on two turbot scuticociliate isolates. Fish and Shellfish Immunology 25, 417424.
Piazzon C., Lamas J. and Leiro J. M. (2011). Role of scuticociliate proteinases in infection success in turbot, Psetta maxima (L.). Parasite Immunology 33, 535544.
Ramos M. F., Costa A. R., Barandela T., Saraiva A. and Rodrigues P. N. (2007). Scuticociliate infection and pathology in cultured turbot Scophthalmus maximus from the north of Portugal. Diseases of Aquatic Organisms 74, 249253.
Rossteuscher S., Wenker C., Jermann T., Wahli T., Oldenberg E. and Schmidt-Posthaus H. (2008). Severe scuticociliate (Philasterides dicentrarchi) infection in a population of sea dragons (Phycodurus eques and Phylopteryx taeniolatus). Veterinary Parasitology 45, 546550.
Saitou N. and Nei M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4, 406425.
Shin P. S., Han J. E., Gómez D. K., Kim J. H., Choresca C. H. Jr., Jun J. W. and Park S. C. (2011). Identification of scuticociliate Philasterides dicentrarchi from indo-pacific seahorses Hippocampus kuda . African Journal of Microbiology Research 5, 738741.
Sievers F., Wilm A., Dineen D., Gibson T. J., Karplus K., Li W., López R., McWilliam H., Remmert M., Söding J., Thompson J. D. and Higgins D. G. (2011). Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Molecular Systems Biology 7, 539.
Small E. B. (1967). The Scuticociliatida, a new order of the Class Ciliatea (phylum Protozoa, subphylum Ciliophora). Transactions of the American Microscopical Society 86, 345370.
Small H. J., Neil D. M., Taylor A. C., Bateman K. and Coombs G. H. (2005). A parasitic scuticociliate infection in the Norway lobster (Nephrops norvegicus). Journal of Invertebrate Pathology 90, 108117.
Smith P. J., McVeagh S. M., Hulston D., Anderson S. A. and Gublin Y. (2009). DNA identification of ciliates associated with disease outbreaks in a New Zealand marine fish hatchery. Diseases of Aquatic Organisms 86, 163167.
Soldo A. T. and Merlin E. J. (1972). The cultivation of symbiont-free marine ciliates in axenic medium. Journal of Protozoology 19, 519524.
Song J. Y., Kitamura S., Oh M. J., Kang H. S., Lee J. H., Tanaka S. J. and Jung S. J. (2009 a). Pathogenicity of Miamiensis avidus (syn. Philasterides dicentrarchi), Pseudocohnilembus persalinus, Pseudocohnilembus hargisi and Uronema marinum (Ciliophora, Scuticociliatida). Diseases of Aquatic Organisms 83, 133143.
Song J. Y., Sasaki K., Okada T., Sakashita M., Kawahami H., Matsuoka S., Kan H. S., Nakayama K., Jung S. J., Oh M. J. and Kitamura S. I. (2009 b). Antigenic differences of the scuticociliate Miamiensis avidus from Japan. Journal of Fish Disesases 32, 10271034.
Song W. (2000). Morphological and taxonomical studies on some marine scuticociliates from China Sea, with description of 2 new species, Philasterides armatalis sp. n. and Cyclidium varibonneti sp. n. (Protozoa: Ciliophora: Scuticociliatida). Acta Protozoologica 39, 295322.
Song W. B. and Wilbert N. (2000). Redefinition and redescription of some marine scuticociliates from China, with report of a new species, Metanophrys sinensis nov. Spec. (Ciliophora, Scuticociliatida). Zoologischer Anzeiger 239, 4574.
Stidworthy M. F., Garner M. M., Bradway D. S., Westfall B. D., Joseph B., Repetto S., Guglielmi E., Schmidt-Posthaus H. and Thornton S. M. (2014). Nondomestic, exotic, wildlife and zoo animals systemic scuticociliatosis (Philasterides dicentrarchi) in sharks. Veterinary Pathology 51, 628663.
Thompson J. C. and Moewus L. (1964). Miamiensis avidus n. g., n. sp., a marine facultative parasite in the ciliate order Hymenostomatida. Journal of Protozoology 11, 378381.
Umehara A., Kosuga Y. and Hirose H. (2003). Scuticociliata infection in the weedy sea dragon Phyllopteryx taeniolatus . Parasitology International 52, 165168.
Whang I., Kang H. S. and Lee J. (2013). Identification of scuticociliates (Pseudocohnilembus persalinus, P. longisetus, Uronema marinum and Miamiensis avidus) based on the cox1 sequence. Parasitology International 62, 713.
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