PARAQUAM Final Activity Report

GENERAL PROGRESS OF THE PROJECT

Due to continuing problems with the availability of Mediterranean amberjack, Seriola dumerilii for experimental work, the focus of the study on sanguinicolid parasites of the genus Paradeontacylix in amberjack was moved to molecular aspects. Various genes of two new species of Paradeontacylix from the Mediterranean were sequenced and analysed together with further samples of other Paradeontacylix species from S. dumerili obtained from Japan. This part of the project has now been published and attracted collaboration with the University of Adelaide, providing samples of Paradeontacylix spp. from Seriola lalandi and Seriola hippos from Australia and New Zealand, which will be used to complete the picture of the worldwide distribution, phylogeny and evolution of Paradeontacylix parasites and their Seriola spp. hosts. The study of the monogenean parasite Zeuxapta seriolae from S. dumerili was completed. For continuation of this line of research, experimental infections were conducted with other monogeneans occurring on gilthead seabream, Sparus aurata and sharpsnout seabream, Diplodus puntazzo. Egg development, larval behaviour and survival rates, as well as infection strategies were investigated in S. aurata, whereas in D. puntazzo the studies were focussed on the distribution of the parasite in the host gills and the related pathology. The planned immunological study of fish infected with monogeneans had to be postponed due to the unexpected early leave of a SR. However, this part of the study has been picked up again recently with the help of an immunologist, who joined the department in October 2007. A new research line focussing on myxozoan parasites of fish has been developed over the last year and a half and results are now being published. One of the studies involves a candidate species for Mediterranean aquaculture, the stripped seabream, Lithognathus mormyrus. Another new line of investigation targets the development of a modern taxonomic framework that will contribute to understanding parasite biodiversity patterns in mullets, which are cultured in the Delta of the Ebro. A similar study is being conducted on metazoan parasites of the annular sea bream, Diplodus annularis (L.), an aquaculture candidate.

Qualitative indicators of progress and success Research results were presented orally and as posters at two international and one national conference. Several articles have been published in peer-reviewed journals and others are still being completed. Due to the interesting results from the sanguinicolid studies, the department was contacted by the University of Adelaide, Australia, for collaboration on this topic. Research in the current Marie Curie project has been amplified regarding parasite and fish species new to the Mediterranean aquaculture. The ER on the project has finished her work satisfactorily and is still publishing results. An SR was recruited but left after setup of a basic immunology lab (6months). Another SR was recruited and is focussing on taxonomical studies. .

PROJECT ACHIEVEMENTS Scientific highlights - Population studies on Paradeontacylix spp. showed that geographic isolation is mirrored in genetic distances leading to speciation of the two parasite species in the Mediterranean (published). One, morphologically similar species from Australia is currently under study to complete the picture of worldwide speciation according to tectonic events. - Monogenean research on aquaculture candidate species of sparids has expanded and infection in a controlled artificial system was established and is being maintained for the investigation of life cycle, biological and pathological aspects of these infections. - A new line of research, i.e. myxozoan parasites was successfully established and resulted in first publications. - In-depth taxonomical studies showed surprising differences between the results of morphological and molecular techniques, changing the known nomenclature of various digenean parasite groups.

Teaching and training activities The researchers of the current project were involved in the setup of an inter-university “Master in Aquaculture” program involving the University of Valencia, the Polytechnic University of Valencia and the Institute of Aquaculture de Torre la Sal (CSIC) as participating scientific establishments and embracing the Erasmus-Socrates Thematic Network “Aquaculture, Fisheries and Aquatic Resource Management” (225991-CP-1- 2005-1-BE-ERASMUS-TN) of the European Union. Webpage: http://www.uv.es/pop/experimentals/aquiculturasp.htm The master was held successfully for the first time in 2006/2007 and thus attracted more than double of the number of students for 2007/2008 with international students from seven different countries. Three PhD students were successfully trained in molecular biology and one PhD student is currently learning immunological techniques. Two master theses with topics embedded in the current lines of investigation and under supervision of the investigators of the present project were conducted and successfully completed in Sept. 2007. The results from the theses are in the process of publication.

Organizer of the “V practical Ictiopathology course for fishfarms. Present health situation of seabream and seabass cultures” with Skretting company and the University of Valencia, Paterna (Valencia, Spain), 22-24 june, 2005.

Dissemination of results

- Peer-reviewed articles published to date and articles in preparation:

Sánchez-García, N., Padrós, F., Raga, J.A., Montero, F.E. (2011). Comparative study of the three attachment mechanisms of diplectanid monogeneans. Aquaculture, 318 (3-4): 290-299.

Alama-Bermejo, G.; Raga, J.A.; Holzer, A.S. (2011). Skoulekia meningialis n. gen., n. sp. (Digenea: Aporocotylidae Odhner, 1912) a parasite surrounding the brain of the Mediterranean common two-banded seabream Diplodus vulgaris (Geoffroy Saint-Hilaire, 1817) (Teleostei: Sparidae): Description, molecular phylogeny, habitat and pathology. Parasitology International, 60: 34–44.

Blasco-Costa, I.; Balbuena, J.A.; Raga, J.A.; Kostadinova, A.; Olson, P.D. (2010). Molecules and morphology reveal cryptic variation among digeneans infecting sympatric mullets in the Mediterranean. Parasitology 137, 287-302.

Holzer AS, Montero FE, Repullés-Albelda A, Nolan MJ, Sitja-Bobadilla A, Alvarez-Pellitero P, Zarza C & Raga JA (2008) Cardicola aurata sp. n. (Digenea: Sanguinicolidae) from Mediterranean Sparus aurata L. (Teleostei: Sparidae) and its unexpected phylogenetic relationship with Paradeontacylix McIntosh, 1934. Parasitology International, 57: 472-482.

Montero, F. E.; Kostadinova, A.; Raga, J.A. (2009). Development and habitat selection of a new sanguinicolid parasite of cultured greater amberjack, Seriola dumerili, in the Mediterranean. Aquaculture, 288: 132-139.

Alama-Bermejo G, Cuadrado M, Raga JA, Holzer AS (2009) Morphological and molecular re- description of the myxozoan Unicapsula pflugfelderi Schubert et al., 1975 from two teleost hosts in the Mediterranean. A review of the genus Unicapsula Davis, 1924. Journal of Fish Diseases, 32: 335-350.

Blasco-Costa, I.; Montero, F. E.; Balbuena, J.A.; Raga, J.A.; Kostadinova, A. (2009). A revision of the Haploporinae Nicoll, 1914 (Digenea: Haploporidae) from mullets (Mugilidae): Dicrogaster Looss, 1902 and Forticulcita Overstreet, 1982. Systematic Parasitology, 72:187–206.

Blasco-Costa, I.; Montero, F. E.; Gibson, D.I.; Balbuena, J.A.; Raga, J.A.; Kostadinova, A. (2009). A revision of the Haploporinae Nicoll, 1914 (Digenea: Haploporidae) from mullets (Mugilidae): Saccocoelium Looss, 1902. Systematic Parasitology. 72:159–186.

Pediguero-Alonso, D.; Montero, F.E.; Kostadinova, A.; Raga, J.A.; Barrett, J. (2008). Random Forests, a novel approach for discrimination of fish populations using parasites as biological tags. International Journal for Parasitology. 38: 1425-1434.

Blasco-Costa, I.; Montero, F. E.; Gibson, D.I.; Balbuena, J.A.; Raga, J.A.; Shvetsova, L.S.; Kostadinova, A. (2008). A revision of the species of Saturnius Manter, 1969 (Digenea: Hemiuridae), parasites of mullets (Teleostei: Mugilidae). Systematic Parasitology, 71: 53–74.

Repullés-Albelda A, Montero FE, Holzer AS, Ogawa K, Hutson KS, Raga JA (2008) Speciation of Paradeontacylix spp. (Sanguinicolidae) in Seriola dumerili with the description of two new species from the Mediterranean. Parasitology International, 57 (3): 405-414.

Pérez del-Olmo A., Fernández M., Raga J.A., Kostadinova A., Poulin, R. (2007) Halfway up the trophic chain: development of parasite communities in the sparid fish Boops boops. Parasitology , 135: 1-12.

Aiken HM, Bott NJ, Mladineo I, Montero FE, Nowak BF, Hayward CJ (2007) Molecular evidence for cosmopolitan distribution of platyhelminth parasites of tunas (Thunnus spp.). Fish and Fisheries, 8(3): 167-180.

Holzer AS, Wootten R. Sommerville C (2007): The secondary structure of the unusually long 18S ribosomal RNA of the myxozoan Sphaerospora truttae and structural evolutionary trends in the Myxozoa. International Journal for Parasitology, 37 (11): 1173-1296.

Yurakhno VM, Ovcharenko MO, Holzer AS, Sarabeev VL & Balbuena JA (2007) Kudoa unicapsula n. sp. (Myxosporea: Kudoidae) a parasite of the Mediterranean mullets Liza ramada and L. aurata (Teleostei: Mugilidae). Parasitology Research, 101 (6): 1671-1680.

Montero FE, Cuadrado M, Padrós F, Crespo S & Raga JA (2007) Cryptocaryon irritans and Enteromyxum leei, two threats for the culture of Diplodus puntazzo in the Mediterranean. Bulletin of the European Association of Fish Pathologists, 27(6): 242 -249.

Holzer AS, Blasco-Costa I, Sarabeev VL, Ovcharenko & Balbuena JA (2006) Kudoa trifolia sp. n. – Molecular phylogeny suggests a new spore morphology and unusual tissue location for a well-known genus. Journal of Fish Diseases 29, 743-755.

Montero FE, Crespo S, Padrós F, De la Gándara F, García A & Raga JA (2004) Effects of the gill parasite Zeuxapta seriolae (Monogenea: Heteraxinidae) on the amberjack Seriola dumerili Risso (Teleostei: Carangidae). Aquaculture 232: 153-163.

- Dissemination of research results at two national and three international conferences:

Alama-Bermejo G, Raga JA, Holzer AS (2008) Myxozoan parasites in two important candidate species in Mediterranean aquaculture: striped sea bream, Lithognathus mormyrus and sharpsnout sea bream, Diplodus puntazzo. Xth European Multicolloquium of Parasitology, Paris, France. ORAL

Holzer A.S., Montero F.E., Repullés-Albelda A., Alvarez-Pellitero P., Sitjá-Bobadilla A., Raga J.A. (2007) A new genus of sanguinicolid blood fluye from Mediterranean gilthead sea bream Sparus aurata L. and its unexpected molecular relationship with other genera of the family. VII International Symposium on Fish Parasites, Viterbo, Italy. ORAL

Holzer, A.S., BAlbuena, J.A., Wootten, R., Sommerville C. (2007) Biology, morphology and molecules of some extraordinary myxozoans and implications from the sinopsis of this information. VII International Symposium on Fish Parasites, Viterbo, Italy. ORAL

Repullés-Albelda A., Montero F.E., Holzer A.S., Raga, J.A. (2007) Speciation of Paradeontacylix spp. (Trematoda, Sanguinicolidae) from Seriola dumerili. Phylogeny of the Paradeontacylix genus. VII International Symposium on Fish Parasites, Viterbo, Italy. ORAL

Blasco-Costa I., Balbuena J.A., Kostadinova A. (2007) Enhanced parasite taxonomy for ecological interactions research: trematodes of mullets (Teleostei, Mugilidae). VII International Symposium on Fish Parasites, Viterbo, Italy. ORAL

Alama-Bermejo G., Holzer A.S., Cuadrado M., Raga J.A. (2007) Morphological, molecular and pathological comparison of two Unicapsula species from the Mediterranean. VII International Symposium on Fish Parasites, Viterbo, Italy. POSTER

Picon-Camacho S.M., Freeman M.A., Holzer A.S., Morris D.J. Shinn A.P. (2007) Description of a new species of Myxobolus Bütschli, 1882 (Myxozoa) from the gills of the common goby Pomatoschistus microps (Teleostei: Gobiidae) in Scottish waters. VII International Symposium on Fish Parasites, Viterbo, Italy. POSTER

Sánchez-Garcia N., Montero F.E., Padrós F., Raga J.A. (2007) Comparison of the attachment mechanism of 3 diplecanid monogeneans: Diplectanum aequans, Lamellodiscus ergensi and Furnestinia echeneis. VII International Symposium on Fish Parasites, Viterbo, Italy. POSTER

Fernández M., Osset E.A., Raga J.A., Kostadinova A. (2007) Metazoan parasite communities of Diplodus annularis (L.) (Sparidae) in the western Mediterranean: geographical variation. VII International Symposium on Fish Parasites, Viterbo, Italy. POSTER

Repullés-Albelda A., Montero F.E., Holzer A.S., Cuadrado M., Raga J.A. (2007) Differences in the pathogenicity of four Paradeontacylix spp. (Trematoda, Sanguinicolidae) from cultured greater amberjack (Seriola dumerili). 13th International Conference of the European Association of Fish Pathologists, Grado, Italy.

Sánchez-Garcia N., Montero F.E., Padrós F., García B., Cerezo J., Raga J.A.(2007) Valoración del efecto patológico del parásito branquial Lamellodiscus ergensi en sargo picudo, Diplodus puntazzo. Comparación con otros monogeneos diplectánidos. XI Congreso Nacional de Acuicultura, Vigo, Spain. POSTER