Conselleria de Pesca e Asuntos Maritimos CENTRO DE INVESTIGACIONS MARIÑAS - CIMA Annual Report 2005

Consellería de Pesca e Asuntos Marítimos Centro de Investigacións Mariñas Annual Report 2005 Technical information

Edit XUNTA DE GALICIA Consellería de Pesca e Asuntos Marítimos. CIMA (Centro de Investigacións Mariñas) (MARINE RESEARCH CENTER) (CIMA)

Coordination CIMA (Centro de Investigacións Mariñas) (MARINE RESEARCH CENTER) (CIMA)

Maquetación IMPRONTA Gráfica s.l NOVA Publicidade Annual Report 2005 Centro de Investigacións Mariñas

Índice

13 1.- INTRODUCTION

17 2.- CENTRO DE INVESTIGACIÓNS MARIÑAS (MARINE RESEARCH CENTER) (CIMA)

19 2.1. STRUCTURE AND FUNCTION

21 2.2. STAFF OF THE CIMA 2.2.1. STAFF EMPLOYED AT THE MARINE RESEARCH CENTER IN VILANOVA DE AROUSA 2.2.2. STAFF EMPLOYED AT THE CENTER FOR MARINE CULTURE IN RIBADEO

26 2.3 FINANCING OF THE CIMA 2.3.1 RUNNING EXPENSES 2.3.2 RESEARCH FINANCING (R+D)

29 3.- SCIENTIFIC ACTIVITY

31 3.1. SUMMARIES OF THE RESEARCH PROJECTS

A.- PROJECTS IN WHICH THE CHIEF RESEARCHER BELONGS TO THE CIMA

3.1.1. AREA OF AQUACULTURE PROJECTS - Optimization of a system for the pre-growth of mollusk spat reared on rafts using alternative energy. - Culture and management of solenaceans (bivalve mollusks). - Reuse of effluents from a seawater fish farm for the grow-out of bivalve mollusk spat - Optimization of the conditioning of the carpet shell clam, Ruditapes decussatus L. - Development of technology for the production and culture of clams. - Promotion of the culture of new sparid species: Sea bream. Pilot trials and technological transfer. - Techniques for the minimization, treatment and use of aquaculture residuals. Subproject A study of the evaluation of fish byproducts in fish farms. Biochemical and microbiological aspects. - Sustainable Harvesting of Ensis (SHARE)- 90.

ACTIONS - Optimization of settlement processes in bivalve larvae reared in mini-hatcheries. An analysis of spat yield with dif- ferent food sources. - An initiative to culture the Pacific oyster (Crassotrea gigas) in different Galician rías. - Restocking of turbot on the Costa da Morte (NW Spain).

9 3.1.2. AREA OF PATHOLOGY PROJECTS - A study of disseminated neoplasia and the pathological alteration known as “large foci with a strong hemocytic reaction” affecting the cockle, Cerastoderma edule, of Galicia. - Commercial solenaceans in Galicia after the Prestige catastrophe: the impact of the oil spill on some biological aspects - Identification of genes as potential indicators of resistance/susceptibilility to diseases in the European flat oyster (Ostrea edulis) and the Pacific oyster (Crassostrea gigas). - Bonamia ostreae life cycle investigations, optimized production of resistant Ostrea edulis spat, and studies of oyster (Ostrea edulis) immune mechanisms.

ACTIONS Optimization of the culture of the flat oyster in Galicia by using quick diagnostic techniques to monitor the parasite Bonamia ostreae. - Development of a genetic selection program to produce a strain of flat oyster, Ostrea edulis L., resistant to bonamiosis.

3.1.3. AREA OF COASTAL OCEANOGRAPHIC PROCESSES

PROJECTS Depuration of Diarrheic type toxins (DSP) in the mussel Mytilus galloprovincialis and amnesic type toxins in the Great Scallop Pecten maximus.

3.1.4. AREA OF MARINE RESOURCES

PROJECTS Introduction of immunodetection techniques to monitor mussel larvae in Galician coastal waters.I. Adaptation of sampling protocols and larva identification. A study of the population stock of the Great Scallop (Pecten maximus) in the Galician Rías.

ACTIONS Geographic Information System oriented towards the management of specific resources (SIGREMAR). Action plan for the recovery of the Bohido zone. Evaluation of larvae and recruitment abundance of mussel spat, Mytilus galloprovincialis, on the Galician coast. Mapping and evaluation of areas and species of interest in the action plan for the recovery of the Lombos del Ulla zone

B.- PROJECTS IN WHICH THE CHIEF RESEARCHER BELONGS TO ANOTHER INSTITUTION. Development of a quick diagnostic method to detect brown ring disease and to discover new bacterial pathogens in the clam. Technology development for a reliable supply of high quality seed in blue mussel farming. Emergency Response to Coastal Oil, Chemical and Inert Pollution from Shipping. (ERCOCIPS).

10 Gradients of biological diversity in intertidal mobile substrates of the Antarctic: Connections between Antarctic and Sub-Antarctic communities (BINTART). Development of elements, tools, protocols for action and an information system for the design of contingency plans in the event of accidental marine spills (CONTINMAR). Subproject: preparation of an operational guide to monitor marine contamination produced by a hydrocarbon spill and its effects on the resources. Genetic evaluation of the phenotypical variants of the European goose barnacle Pollicipes pollicipes across its geographical distribution. Byproducts from aquaculture in Galicia. Carbon flows through Plankton in Subtropical Oligotrophic environments: A Lagrangian Approach (CARPOS).

97 3.2 THESES AND RESEARCH WORKS (DOCTORAL PROGRAM) CARRIED OUT AT THE CIMA

111 3.3 ADVISORY SERVICES FOR THE SECTOR AND THE ADMINISTRATION

115 3.4 SCIENTIFIC AND TRAINING ACTIVITIES CARRIED OUT BY THE STAFF 3.4.1. ORGANIZATION, CHAIRMANSHIP AND PARTICIPATION IN SCIENTIFIC MEETINGS 3.4.2. DIRECTION OF DOCTORAL THESES AND INTERN TRAINING 3.4.3. COURSES GIVEN 3.4.4. PARTICIPATION ON EXAMINING BOARDS TO JUDGE DOCTORAL CANDIDATES 3.4.5. EDITION OF SCIENTIFIC PUBLICATIONS 3.4.6.REVIEW OF ARTICLES IN SCIENTIFIC JOURNALS 3.4.7.PATENTS

128 3.5 STAFF TRAINING 3.5.1 VISITS TO OTHER CENTERS 3.5.2 COURSE ATTENDANCE

130 3.6 COLLABORATION AGREEMENTS

131 3.7 OUTSIDE VISITORS

132 3.8 PUBLICATIONS 3.8.1 ARTICLES IN JOURNALS LISTED IN THE SCI 3.8.2 ARTICLES IN JOURNALS NOT LISTED IN THE SCI 3.8.3 CONTRIBUTIONS TO BOOKS

139 4. SCIENTIFIC AND TRAINING FORUMS ORGANIZED BY CIMA 141 4.1 SEMINARS

143 5. ANNEX. Abbreviations used.

11

1. INTRODUCTION

As in previous years, the activity report presents The research work combines scientific production the work done and the goals accomplished through published in high-impact journals with a more the research and assistance services carried out at direct, immediate approach, related to productive the Centro de Investigacións Mariñas (Marine systems. A total of 57 presentations were made at Research Center) or CIMA in 2005. different scientific meetings of different types (con- ferences, seminars, forums, workshops, etc.) and The different sections into which this report is areas: 8 presentations at the regional level; 22 at divided will attempt to offer an overview of the national venues; and a total of 26 papers were pre- work carried out at the Research Centers of the sented at international meetings. In terms of the dis- Local Ministry for Fisheries and Maritime Affairs semination of the research work, this year 18 articles located in Vilanova de Arousa and Ribadeo. were published in top-ranking, high-impact scientif- ic journals and 14 papers were published in national Scientists at the CIMA headed a total of 13 research journals, not to mention contributions to research projects and 9 research actions. The former books (4). were awarded and financed through public notice on different levels: regional (Galician Program IDIT); This high level of investigative work and scientific national (national R+D Program, JACUMAR, etc.) production would not be possible without the con- and European (AquaReg, INTERREG, 5th and 6th EU tribution of the temporary workers (both scientists Framework program, etc ). The research actions, and technicians) who are involved in the different however, are aimed at responding to marine research projects and actions and who, in most resource management problems that may be detect- cases, have been trained in our research centers. ed by the Galician Fisheries Administration and are financed with funds from the Local Ministry for In keeping with the paragraph above, an impor- Fisheries and Maritime Affairs. The center has also tant role in terms of training is also carried out by participated in and collaborated on a total of 9 proj- the researchers through the direction of doctoral ects in which the chief researcher belongs to anoth- theses ( 10 ) and technical training ( 9) in different er institution, either national (IEO, Universities, fields of expertise (aquaculture production, pathol- CETMAR, etc.) or international: with Ireland collabo- ogy, advanced diploma in systems...); they are also rating on a project under the 6th European Union involved in practical training and in the direction of Framework Program to improve mussel spat and Master’s theses (2) as well as vocational on-the-job another EU project which includes the participation training in companies, etc. of several Atlantic countries (Ireland France, Portugaland others), dealing with the protocols for While CIMA’s goal is not to be a self-financing action and response to contamination events caused entity “per se”, it is interesting to note that the by accidental spills from vessels. funds obtained for research account for 42% of CIMA’s running expenses and of these funds 72% The section on advisory services and assistance come from outside projects awarded through public was prepared on the basis of a large number of notice. reports issued. These reports were generally reques- ted through the Directorate General of Resources CIMA’s goal is to implement research in keeping for the Fishermen’s and Harvesters’ Associations, with the demands and needs of Galicia. To this end, INTECMAR, etc. Members of the CIMA staff also all of the researchers must be fully involved, using assiduously attended meetings and represented the their skills to develop projects and lines of investiga- Galician Administration in different types of com- tion within a general framework of quality and missions, mainly of a technical nature, where the applied research work that is fundamentally produc- opinions and criteria for action of the fisheries tive and whose aim is to promote the service and administration are communicated in the different sustainable development of the live marine areas of competence and interest to the sector. resources in Galicia.

Fátima Linares Cuerpo General Directora of Innovation and Fisheries Development

15

2. CENTRO DE INVESTIGACIÓNS MARIÑAS (MARINE RESEARCH CENTER)

CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER

2.1. STRUCTURE AND FUNCTION

The Centro de Investigacións Mariñas (CIMA), Area of Aquaculture. The objective of this area is belongs to the Local Ministry for Fisheries and to develop and improve methods of rearing species Maritime Affairs, and comprises the Marine considered to be marketable. There are two basic Research Center of Corón in Vilanova de Arousa lines of research: mollusk culture and fish farming. (Pontevedra) and the Marine in Ribadeo (Lugo). The basic function of the CIMA is to carry out research In the line of investigation aimed at mollusks, a aimed at achieving the rational management of number of research projects are currently underway. renewable marine resources within the geographic The projects are aimed at improving spat produc- scope under the jurisdiction of the Administration tion, mainly in hatchery-reared clams, and they of Galicia. entail analyses of nutritional aspects using microcap- sules in the larval and post-larval stages in addition Scientific activity is divided into four areas: to determining the methods of selection to be used in the hatchery to improve spat quality. The study of Area of Marine Resources. The objective is to gain reproduction, the procurement and culture of spat knowledge on the biology, ecology and population of commercial solenidae (razor clams) is part of dynamics of marine species having commercial inter- another project underway. Another aspect is the est in order to improve the management of these analysis of the outdoor culture technology (seed natural resources. There are currently two lines of beds for the grow-out of clams on rafts and long- research underway. line systems for mussels).

The first deals with the larval ecology of marine The research on fishes includes a number of proj- invertebrates examining both the spatial and tem- ects that study species which currently offer the poral distribution of the larval stages in the coastal most promising perspectives for the “standardiza- zone as well as their behavior. This information is tion” of their culture in the near future: the aimed at determining the relation that exists blackspot sea bream (nutritional aspects in the dif- between the abundance and recruitment of larvae, ferent stages of culture and outdoor culture in as well as the possible influence of environmental cages) and the sole (evaluation of diets for parent conditions on both phenomena. These studies will stock and larvae). In 2005, a program was started to attempt to explain the great spatial-temporal vari- restock marine species The first action consisted of ability in recruitment observed in most marine inver- the release of turbot fry with the main goal being to tebrates of commercial interest and to design strate- recuperate the stocks of several species on the gies that will help increase recruitment. Galician coast. Various aspects on the culture of the octopus are The other line of research focuses on the develop- also being studied, such as the effect of food consist- ment and maintenance of a geographic information ing of different prey on octopus paralaravae. system to be used as a tool in the management of shellfish species. All the available information on Area of Pathology. The objective is to study the the natural beds in Galicia and the shellfishery activ- pathological alterations affecting bivalve mollusks ity carried out in these areas is stored in the data of commercial interest in order to set up strategies bases of this system. The analyses carried out using that will effectively combat these alterations lead- this system are designed to provide solutions to ing to a reduction in the pathological effects. The problems related to shellfish management and to lines of research include the following aspects: successfully obtain a sustainable and lucrative exploitation..

CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS Memoria CIMA 2005 19 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

Effects on natural populations and cultures of ery and shellfishery resources in Galicia. There are bivalve mollusks. Effects on survival, growth (ener- currently two lines of research, both of which are getic physiology), quality and reproductive capacity. related to toxic episodes caused by phytoplankton. Distribution and incidence on the Galician coast. Etiology. Identification and characterization of The first line of research is aimed at the develop- pathogens. ment and application of techniques that are alter- Diagnostic techniques. native and/or complementary to microscopy in Temporal dynamics of pathological alterations/ order to study the composition and dynamics of the pathogens. phytoplankton responsible for harmful algale Influence of environmental conditions. blooms. Life cycle of pathogens. How they are transmitted. Culture of pathogens. The second line involves the study and modeling Virulence factors. of the accumulation of toxins from phytoplankton Interaction with the immune system of bivalve in bivalve mollusks and the design of procedures to mollusks. accelerate the depuration of these toxins. Combat strategies: production of resistant stocks. Prophylaxis. Therapeutic treatments. The CIMA is also involved in training new researchers and laboratory technicians with the Area of Coastal Oceanographic Processes. The temporary placement of these people at the Center objective of this area is to shed light on the aspects itself and by teaching courses in doctoral programs. of oceanography related to the production of fish-

Centro de Investigacións Mariñas Pedras de Corón s/n. Apartado 13. 36620 Vilanova de Arousa (Pontevedra). Spain Tel.: +34 986 50 01 55 +34 986 50 01 61 Fax: + 34 986 50 67 88 e-mail: [email protected]

Centro de Cultivos Mariños Muelle de Porcillán s/n. Apartado 94. 27700 Ribadeo (Lugo). Spain Tel..: +34 982 12 81 00 +34 982 13 04 92 Fax : +34 982 13 03 91 e-mail: [email protected]

web: http://www.cimacoron.org

20 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

2.2. PERSONAL DEL CIMA

PERSONAL DEL CENTRO DE INVESTIGACIÓNS MARIÑAS (CIMA), EN EL AÑO 2005

CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 21 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

2.2.1. STAFF EMPLOYED AT THE MARINE RESEARCH CENTER OF VILANOVA DE AROUSA.

Director: Dr. Alejandro Guerra Díaz [email protected] Manager: D. José Barreiro Piñeiro [email protected]

Researchers Ms. Fátima Linares Cuerpo 3 Dr. Elvira Abollo Rodríguez [email protected] [email protected] Dr. Mª del Carmen López Gómez Dr. José L. Alonso Fernández de Landa [email protected] [email protected] Dr. José Molares Vila Dr. Juan Carlos Blanco Pérez [email protected] [email protected] Dr. Jaime Montes Pérez Dr. Mª Jesús Carballal Durán maria. [email protected] [email protected] Ms. Mª José Outón Caamaño Dr. Sandra Casas Liste4 [email protected] Mr. Antonio Cerviño Eiroa Ms. Carmen Pérez Acosta [email protected] [email protected] 1 Mr. Alberto de Coo Martín Dr. Mª Eugenia Rodríguez Moscoso [email protected] [email protected] 2 Dr. José Miguel Fuentes González Dr. Adoración Sánchez Mata [email protected] [email protected] Mr. Antonio García Fernández Mr. Ignacio Santos Piñeiro [email protected] [email protected] Dr. Alejandro Guerra Díaz Dr. Antonio Villalba García [email protected] [email protected] Dr. Salvador Guerrero Valero Dr. Manuel Zapata Gago [email protected] [email protected]

1 Staff currently carrying out their duties at another administrative unit. 2 Parga Pondal Program 3 Ramón y Cajal program 4 Permanent staff associated with research projects.

22 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

Post-Graduate Fellows Head of the Administrative Department Ms. Pilar Comesaña Lestayo Mr. Gonzalo García Paz Ms. Seila Mª Díaz Costas [email protected] Ms. Silvia Lorenzo Abalde Ms. Laura Domínguez Pérez Librarian Ms. Patricia Mirella da Silva Ms. Mª Pilar Fraguela Villar Ms. Aida Mauriz Pereira [email protected]

Lab Analysts Senior Administrative Staff Mr. Juan Antonio Fariña Iglesias Ms. Lucinda Caamaño Pérez Ms. Mª Mercedes Miranda Bamio [email protected]

Lab Technicians Administrative Assistants D. Ramón Giráldez Rivero Mr. José Lago Torrado Ms. Mª Victoria Gregorio Chenlo [email protected] Ms. Mª Carmen Mariño Cadarso Mr. Pablo Ventoso Padín Ms. Iria Santamaría Búa4 [email protected] Ms. Obdulia Varela Manso4 Ancillary Employees D. Luis Manuel Agra Carregal Lab Assistants D. José Manuel Martínez Crespo Ms. Emilia Campaña Ferro Ms. Mª José Cores González Security Guard Ms. Mª Isabel Meléndez Ramos Ms. Mª Josefa Barrosa Vázquez Ms. Elena Penas Pampín Cleaners Ms. Ramona Maneiro Lamas 5 Vocational Interns 6 Ms. Carmen Angueira Souto Ms. Mª Josefa Gallego Cebro Ms. Rosana Rodríguez Díaz 5 Mr.. Pablo Cachafeiro Diéguez 6 Ms. María Otero Otero Ms. Nerea Soto Fraga Ms. Mercedes Rodal Mayo 5 Ms. Teresa Varela Soñora 6 General Maintenance Mr. Ramón García García

5 Renounced grant/contract. 6 Substitutes for the above .

CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 23 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

2.2.2. STAFF EMPLOYED AT THE CENTER FOR MARINE CULTURE. RIBADEO

Researchers Vocational Interns D. Fiz da Costa González 4 Ms. Sonia Rábade Uberos 5 [email protected] Mr. Diego Rial Conde 6 Ms. Aurora Fernández Álvarez [email protected] General Maintenance Dr. Dorotea Martínez Patiño Mr. José Mª Loureiro Barcón [email protected] [email protected] Ms. Susana Nóvoa Vázquez [email protected] Senior Administrative Staff Ms. Justa Ojea Martínez Ms. Concepción Neira Páez [email protected] [email protected]

Post-graduate fellow Administrative Assistant Ms. Ana Cerviño Otero Ms. Ana Cruzado Estévez [email protected] [email protected]

Lab Analyst Ancillary Employee D. Luis Manuel Álvarez Llamas Ms. Mercedes Loureiro Rodríguez [email protected] Cleaner Lab Technician Ms. Carmen Fernández Goás Ms. Mª Victoria Bouzamayor Yáñez [email protected]

Lab Assistants Ms. Mª Josefa Cotarelo Jardón [email protected] Mr. Francisco Javier Páez Pérez [email protected] Ms. Mercedes Ruiz Misioné [email protected]

24 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

2.2.3. TEMPORARY STAFF, ASSOCIATED WITH THE RESEARCH PROJECTS CARRIED OUT AT THE CIMA7.

Researchers Ms. Mª Consuelo Darriba Santiago Dr. Susana Darriba Couñago. Mr.. Isidro Fernández Abuín Dr. Asunción Hermida Cao Ms. Helena Martín Sánchez Dr. José Manuel Parada Encisa Ms. Gema Pazos Sieira Mr. David Iglesias Estepa Ms. Genma Martínez Verde Ms. Mercedes Rodal Mallo Technicians Ms. Rosana Rodríguez Díaz Ms. Alicia Alvarez Padin Ms. Marta Andrade García. Computer Technicians Mr.. Pablo Carreira Vazquez Ms. Mª Loreto Rodriguez Ruibal Ms. Ana González Rodríguez Mr. Alberto Crego Mata

7 Temporary staff associated with research projects or actions carried out by CIMA, employed through agreements with their respective institutions.

CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 25 CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

2.3. FINANCING OF THE CIMA

2.3.1.- RUNNING EXPENSES OF THE CIMA

2.3.2.- PROCUREMENT OF FUND FOR RESEARCH (R+D).

26 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005

3. SCIENTIFIC ACTIVITY

SCIENTIFIC ACTIVITY

3.1. SUMMARIES OF THE RESEARCH PROJECTS

A.- PROJECTS IN WHICH THE CHIEF RESEARCHER BELONGS TO THE CIMA

3.1.1. AREA OF AQUACULTURE

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Optimization of a system for the pre-growth of mol- Cerviño Eiroa, A. lusk spat reared on rafts using alternative energy. RESEARCHERS: START: September, 2002 CIMA: De Coo Martín, A.; García Fernández, A. FINISH: September, 2005 USC: Bao Iglesias, M.

FINANCIAL ASSISTANCE: RESEARCH SUPPORT: Plan Galego de Investigación e Desenvolvemento Fariña Iglesias, J. A. Tecnolóxico - (Galician Program for Research and Technological Development) (PGIDT) PGIDT02RMA50102PR

SUMMARY The research project entitled “Optimization of a with no verifiable differences being found, since if system for the pre-growth of mollusk spat reared on differences did exist at any given moment depend- rafts using alternative energy” was completed in ing on the direction of the prevailing winds at the 2005. time, at the end of the year they would tend to be compensated for and even out. Over the course of the last year an analysis was planned to be carried out on the basis of the data The results obtained from the harnessing of solar obtained in both the monitoring activity and energy using photovoltaic panels placed on the raft through the different tests done, and may be sum- indicate that in order to reap maximum benefits, it marized in the following aspects: would be necessary to have a greater surface area, and still, it would not be highly efficient in all of the Production capacity and the monitoring of the seasons of the year. Therefore, we would propose alternative energy systems planned to be used on the use of wind power as a feasible complement. rafts. Grow-out of mollusk spat. To set up spat holding systems using forced circu- Safety and Environmental protection. lation, it is necessary to take into account the effects of the wave action on the spat. The systems that From the standpoint of production, we analyzed have proved to work the best are those in which the the results from the production of wind and solar spat is submerged at a depth ranging between 1 power, as well as energy from seawater currents. and 2 meters, which in terms of industrial practice, These results were confirmed with a wind generator, makes handling difficult and also limits holding which indicated that it is possible to use between 55 capacity. This would be only considered appropriate and 65 % of the hours annually (depending on the for very small-sized spat with low individual body year) to harness energy in different areas of the Ría weights, which means that with a reduced volume de Arousa. During the period from Sept./04 to in biomass held on the rafts, it is possible to main- Sept./05, an average percentage of 58.34% was tain a large number of individuals per grow-out recorded, which is equivalent to a total of 5109.72 unit. useable hours during this period. We have also carried out grow-out tests with mol- lusk spat, reducing the initial size in systems that do During the analysis of the capacity to produce not use forced circulation. The technology of the energy based on the possible location of the raft in grow-out system in mini-bags held horizontally at the different raft parks situated in the Ría de Arousa different levels in a metallic cage was used. The first area, wind was measured in other parts of the Ría, grow-outs were started in 1 mm mesh bags, which

CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 31 SCIENTIFIC ACTIVITY

would allow us to begin the grow-out cycle with a This method of water procurement is more homo- spat size of 1500 microns. geneous throughout all of the biomass, which reduced the formation of tails. The growth and survival results were very good, with mean growth rates (Gm) reaching 200 in the In the section on safety, we would like to point pullet carpet shell clam (Venerupis pullastra), 150 in out that it is necessary to comply with the regula- the short necked clam (Ruditapes philippinarum) tions regarding electrical installations, especially and 100 in the carpet shell (Ruditapes decussata), under circumstances where the saline environment with survival rates of over 95% during the grow-out increases the risks. It was necessary to install a stage. lighting rod and different safety mechanisms to protect both the equipment and the people who The clams held in bags are able to take in water, carry out different tasks while working on the raft and with it the food and oxygen they require, during the year. directly form the outside of the bag, optimizing the use of the available seawater, which means that there is a maximum load capacity. Spat loss due to handling or other causes is minimized.

32 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Culture and management of solenaceans Guerra Díaz, A. (bivalve molluscs). Coordinated project

START: November, 2002 RESEARCHERS: FINISH: November, 2005 CIMA: Da Costa González, F.; López Gómez, C.; Martínez Patiño, D; Montes Pérez, J.; Nóvoa FINANCIAL ASSISTANCE: Vázquez, S.; Rodríguez Castro, V. Plan Nacional de Cultivos Marinos. Junta Asesora de CETMAR: Darriba Couñago, S. Cultivos Marinos - (National Program for Marine Delegación de CPAM de A Coruña: Culture – Marine Culture Advisory Board) Catoira Gómez, J.L. (JACUMAR) CCMM: Fernández Conchas, R.; Álvarez Álvarez, C.

RESEARCH SUPPORT: Meléndez Ramos, M.I.; Miranda Bamio, M.; Penas Pampín, E.

SUMMARY In Spain there are three important commercial conditions: one in the Ría de Vigo (Barra – Cangas species of razor clams: (Ensis arcuatus), (Ensis siliqua) beach) and another on A Costa da Morte (Fisterra – and (Solen marginatus). The aim of this project, Ría de Corcubión). The results show that in both coordinated by Galicia and including the participa- locations the animals undergo a sexual rest period tion of four Autonomous Communities: Galicia, during the entire summer and autumn period, while Asturias, Cantabria and Andalucia, is to analyze the spawning is concentrated in spring (April-May)- In state of this shellfish resource on the coast of Spain. the bed located in Barra, a small percentage of indi- Below is a summary of the results obtained by the viduals was found to spawn until early July. group working in Galicia for each line of investiga- tion: Solen marginatus was studied in 2003, 2004 and 2005 in the en la Ría de Arousa (Mañóns-Cabo de Reproduction.- A comparative study on the repro- Cruz beach), in a bed where the population has ductive cycle of the three species of solenaceans been drastically reduced in recent years. In 2005, the (Ensis arcuatus), (Ensis siliqua) and (Solen margina- study was extended to include a comparison of this tus), selecting for each one, two beds having differ- bed with the bed having the highest production of ent oceanographic conditions. this species in Galicia, located in Redondela, (San Simón Cove-inner Ría de Vigo). The results obtained The cycle of Ensis arcuatus was studied in a subti- show a similar strategy, with a sexual rest period dal bed in the outer area of the Ría de Vigo (Rodas from July to December and the spawning period beach – Cíes Islands) and from a low intertidal zone being concentrated from May-June. in the inner Ría de Arousa (Meáns -Cambados). The results obtained would indicate a similar behavior in Pathology.- Pathological analyses were performed the two locations during the two years of study from January, 2003 until May, 2005 every three (2003 and 2004). months, to determine the pathological condition of the following: Ensis arcuatus in the Ría de Vigo The cycle begins in autumn and consecutive (Rodas beach Cíes Islands),and in the Ría de Arousa spawning takes place during winter and spring. This (Meáns-Cambados); Ensis siliqua in the Ría de Vigo species undergoes a sexual rest period during the (Barra-Cangas beach) and on A Costa da Morte summer and part of the autumn. (Lagosteira beach- Fisterra); and Solen marginatus in the Ría de Arosa (Mañóns-Cabo de Cruz beach). Ensis siliqua was studied in two subtidal beds in The main parasites observed were ciliates in the the far coastal zone and in different environmental gills, bacterial bags, the gregarine Nematopsis sp., coccidia, turbellarians, copepods, trematodes and

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organisms of the rickettsian/chlamydias type. Special Experiments with the razor clams E. arcuatus and attention should be paid to the presence of E. siliqua did not provide satisfactory results owing Marteilia sp. detected in Solen marginatus. to problems having to do with the deterioration of Alterations in the tissue were caused, in some cases, the culture equipment. New experiments are being by the presence of infiltrations, granulocytomas and carried out with these species. tissual necrosis. Depuration.- The system commonly employed for Hatchery production.- Different techniques were the depuration of bivalve mollusks in commercial perfected: the correct holding and conditioning of facilities presents a number of different problems the parent stock, inducement of spawning, the for solenacean species. The weak muscle system hatchery culture of the larvae, postlarvae and spat causes the valves to open quickly after they have of the three species. It is more difficult to obtain been buried in the sand for a long time, which accel- spawning in the two species of Ensis, than in S. mar- erates their deterioration. Our aim was to find ginatus. depuration conditions for the three species that would be easy to apply with the routine practices It was concluded that the most appropriate condi- used in commercial treatment systems that would tioning of E. arcuatus was obtained by keeping the guarantee the optimum quality of the individuals seawater temperature within the range of the out- after the completion of the process. door temperature, changing the water level, includ- ing brief drying out periods, to simulate the tides. It was found that if the razor clams (previous According to the conditioning experiments carried selection of undamaged specimens) are tied into out on the 3 species in the different seasons of the bunches of 15-20 individuals, arranged horizontally, year, we would recommend that conditioning be they will be able to withstand the depuration started during the initial stages of gametogenesis, process, resulting in survivals of 100% and with opti- which would make it possible to advance spawning mal marketing conditions. The multiple experiments by 1 or 2 months. Larval culture presented no prob- carried out demonstrate that it is not possible to lems and Solen, which has a short larval period establish depuration times for each species, since (from 7-10 days), exhibited the highest survival there is a high variability depending on the season, rates (>30%). In the postlarval stage, the survival initial individual load, the load of the treatment sys- rate dropped (< 5 % between 1 and 3 mm). It is nec- tem, etc. In any event the main objective of the proj- essary to determine the most suitable method for ect was fulfilled, defining a depuration system and the grow-out of the spat. holding conditions for individuals that will allow them to withstand the process without any damage Outside spat culture.- Spat was sown for two pur- that would in any way either jeopardize or impair poses -to restock natural beds and to monitor spat in their commercial value. controlled areas to determine growth and survival. Monitoring in controlled areas was carried out in Information on the resource.- Geographic Infor- cages constructed with stainless steel rods (10 mm) y mation System. The new web page of the Ministry measuring 50 x 50 x 50 cm., and these were covered of Agriculture, Fisheries and Food is now available. with a 6 mm mesh having an opening at the top. It is a site with information on the bibliography rel- ative to solenacean species appearing in Europe and Specimens of S. marginatus, measuring 19.1±1.92 the latest data on the harvesting of solenaceans in mm (equivalent to 4 months of age), reached a size Spain by Autonomous Community: of 38.53±3.89 mm and weight of 1.56±0.46, 8 http://www.mapya.es/jacumar/ months after being sown. After 19 months of life, the accumulated survival was 70 %, with a size of 67.01±3.32 mm. The experiment is planned to con- tinue until commercial size is reached (80 mm).

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Reuse of effluents from a seawater fish farm for the Guerra Díaz, A. grow-out of bivalve mollusk spat. COMPANY: START: 2004 INSUIÑA, S. L. (Grupo Pescanova). FINISH: 2005 Researcher in charge: Fernández Casal, J.

FINANCIAL ASSISTANCE: RESEARCHERS: Plan Galego de Investigación e Desenvolvemento CIMA: Martínez Patiño, D; Nóvoa Vázquez, S. Tecnolóxico - (Galician Program for Research and IGAFA: Andrés Rivas, M.C.; Lastres Couto, M. Technological Development) (PGIDT). Marine Resources Program. RESEARCH SUPPORT: PGIDIT04RMA013E Loureiro Barcón, J.M.; Miranda Bamio, M.; Ruiz Cruz, A.

SUMMARY Three grow-out trials were carried out with three using the combined resources of the natural envi- clam species (carpet shell, Ruditapes decussatus; ronment and the hatchery itself. pullet carpet shell, Venerupis pullastra and short necked clam, Tapes philippinarum), during three In winter all three species presented an irregular seasons to determine the feasibility of their growth growth pattern. The pullet carpet shell clam grew throughout the year. In the first experiment three more than the other species. This was partially due types of grow-out densities were also tested to iden- to the fact that its initial size was larger and that the tify the one most suitable to the characteristics of temperature interval of this experiment (between the facility (both physical and environmental). 15.2 and 6.8oC) was within the range at which the ingestion rate of this species reaches its highest val- The autumn-winter and spring experiments were ues. carried out with spat from the three species, while the summer trial was only conducted using the pul- Despite the low temperatures (below 10 oC on let carpet shell and short necked clam. Mean sizes average during the experiment), the three species and weights fluctuated: from 4.5-7 mm and 16-69 underwent growth during a period in which the mg in the short necked clam, 4.1-4.6 mm and 20-21 concentration of chlorophyll “a” was not greater mg in the carpet shell, and lastly 5.8-7.9 mm and 26- than 1 Ìg/L in the natural environment. Growth was 87 mg in the pullet carpet shell. supplemented by a contribution of organic matter from effluents. Results: Of the three densities tested, it was the lowest In spring the pullet carpet shell clam reached a density that exhibited the best results with the short size of 10 mm, in 22% less time than in winter. It is necked and pullet carpet shell clams. The growth of important to note that this growth period could the carpet shell clam was less linear in both weight even be lower since the spat, in this case, suffered and length, which may be attributed to the wide high mortalities at the beginning of the experiment range of sizes found in this species. On the basis of owing to circumstances beyond the control of the an analysis of the results of standard density (6.6 experiment (25 % during the first 15 days). The car- 2 kg/m ) in the three species, it was observed that the pet shell and short necked clam had exhibited the growth of the clams was similar to their growth at same behavior as during the winter period, except lower densities, and that they reached the desired for the fact that in only 80 days they reached the size between 20 and 30 days later than when reared size they attained in winter after 133 days. This in lower densities. Densities used in traditional cul- growth pattern is largely due to the origin and qual- 2 tures do not usually exceed 2.5 kg/m . This system ity of the spat. proposed here increases the grow-out capacity by

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The grow-out period in summer was the shortest Spat quality is a determining factor for the effi- of the whole year – the clams reached a size of 10 cient functioning of the system. mm in 77 days. To optimize the grow-out for each hatchery, it is Conclusions: necessary to establish the relationship between spat This system, -easy to handle and efficient- is suit- biomass versus the characteristics of the facility. able for the grow-out of clam spat. The minimum size must not be less than 3 mm.

36 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Optimization of the conditioning of the carpet shell Martínez Patiño, D. clam, Ruditapes decussatus L. Project coordinated by Sánchez López, J.L. (USC). START: August, 2002 FINISH: August, 2005 RESEARCHERS: CIMA: Nóvoa Vázquez, S.; Ojea Martínez, J. FINANCIAL ASSISTANCE: Plan Galego de Investigación e Desenvolvemento RESEARCH SUPPORT: Tecnolóxico - (Galician Program for Research and Alvarez Llamas, L. M.; Bouzamayor Yáñez, M. V.; Technological Development) (PGIDT) Cotarelo Jardón, M. J.; Loureiro Barcón, J. M.; Páez PGIDIT02RMA23501PR Pérez, F.J.; Ruíz Misioné, M.

SUMMARY This project was presented as a project carried out Two experiments were conducted to achieve in coordination with the team of the Aquaculture objective 1.2: Institute of the University of Santiago de Compostela (biochemistry and molecular biology First conditioning. Three different diets were test- group) and a team from the CIMA which belongs to ed under the same temperature regime – diets com- the Xunta de Galicia (Regional Government) (the prising 3, 6 and 9% microalgae dry weight/clam dry group from the Center for marine Culture in weight, kept at a temperature that was gradually Ribadeo). increased from environment temperature to a max- imum of 20ºC. The experiment began with clams in The main goal of this project was to optimize the the initial stage of gametogenesis. The best results conditioning process of the parent stock of the car- in terms of the parameters studied (mortalities, con- pet shell clam, Ruditapes decussatus. This goal was dition indices biochemical composition of the pursued and broken down into the following partial gonads, maturity stage and spawnings) were found objectives: using the 6%, diet although the biochemical com- position was similar in all three. 1.1.-The combined effects of the photoperiod, temperature and diet on conditioning in autumn Second conditioning. A combination of two diets: (gametogenic rest stage). 3 and 6% and two temperature regimes: one grad- ed, increasing 0.5ºC per week and the other at a 1.2.- Interaction between temperature and diet constant temperature of 20ºC from the beginning during winter conditioning (initial stage of the of the experiment. development of gametogenesis). The best results in terms of gonad development 1.3.- The relationship between diet and the level were found in clams that were fed a double diet at of water renewal during conditioning in spring a constant temperature. All of these clams reached (intermediate stage of gametogenic development). maturity. In contrast, the poorest results were obtained using the normal diet and a graded tem- 1.4.- A study of the possible effect of the involu- perature regime. Under these conditions only 25% tion of gametogenic development using a winter of the parent stock reached maturity. As regards the photoperiod during spring conditioning. other two conditions, the one with the poorest results was the double diet with a graded tempera- The subproject carried out by the team at the ture regime, which still showed 8% of the clams CIMA-Ribadeo dealt with objectives 1.2 and 1.3. starting gametogenesis. Lastly, the 86% of the clam population fed a normal diet at a constant temper- ature reached maturity.

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In general the best results were found using the biochemical composition are somewhat better. In double diet at a constant temperature, although, terms of the maturity stage and spawning success, depending on the parameter studied, there are cer- all three showed similar results. tain variations. For example, the fundamental factor in the condition index is the diet, since the best In summary, we may conclude that the tempera- results were found using the double diet, at both ture factor is more important in gonad development constant temperatures and those undergoing a than the amount of food administered. As long as gradual increase. The most influential factor in the the quantity of microalgae is not a limiting factor, mortalities of the parent stock is the temperature. we consider that it should be greater than 3% of the Clams subjected to a gradual temperature increase parent stock dry weight. The best results were found exhibited lower mortalities. As regards the biochem- with diets of around 6%. Diets higher than this ical composition, the best results were seen in clams would not be worth the expense in phytoplankton held at a constant temperature rather than a grad- production. ed regime. Spawning took place under all the condi- tions except the one using the normal diet with a The water temperature regime having the least graded temperature. The first spawnings were col- impact on parent stock mortality was the one that lected a month and a half after conditioning was increased gradually during the conditioning process, begun. although, if the temperature drops from 20 to 18ºC, problems with mortality are substantially reduced For Objetive 1.3.-. Conditioning was carried out and, moreover, the constant temperature speeds up using three different water renewal rates: 1/3, 1/5 the maturation process. The ideal situation is to hold and 1/7 of the total volume of the tank in one hour. the parent stock at a temperature similar to what is found in the natural environment during the first This conditioning, which should be done in spring, few days in the hatchery so they will be able to was carried out in winter owing to the difficulty we adapt: The temperature should later be increased had obtaining parent stock. Therefore, instead of and kept constant at 18ºC±1. being in the intermediate stage of gametogenic development, the clams arrived in the initial devel- The level of water renewal in the tank does not opmental stages. In this conditioning experiment, have any noticeable effect on the conditioning similar results were obtained for the three rates. processes. Of all the rates studied, the most suitable Renewal rate 1/5 is the one that presented the low- would be to renew 1/5 of the tank volume every est mortality and the condition indices as well as the hour per kilogram of clam live weight.

38 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Development of the technology for the production Martínez Patiño, D. and culture of clams. Coordinated project

START: January, 2005 RESEARCHERS: FINISH: December, 2007 CIMA: Carballal Durán, M.J.; Cerviño Eiroa, A.; de Coo Martín, A.; García Fernández, A.; FINANCIAL ASSISTANCE: Guerra Díaz, A.; López Gómez, M.C.; Plan Nacional de Cultivos Marinos. Junta Nacional Nóvoa Vázquez, S.; Ojea Martínez, J. Asesora de Cultivos Marinos. (National Program for IGAFA: Andrés Rivas, M.C.; Lastres Couto, M.A. Marine Culture – Marine Culture Advisory Board) (JACUMAR) RESEARCH SUPPORT: Alvarez Llamas, L. M.; Bouzamayor Yáñez, M. V.; Cotarelo Jardón, M. J.; Fariña Iglesias, J.A.; Meléndez Ramos, M.I.; Miranda Bamio, M.; Páez Pérez, F.J.; Penas Pampín, E.; Ruíz Misioné, M.

INTERN Cerviño Otero, A.

SUMMARY This is a nationwide project involving the partici- Larval and postlarval culture. All three species pation of several groups from the CIMA in coordi- were cultured. A similar protocol was used for the nation with four other autonomous communities, three types of clams. Larval culture in 500 liter tanks whose objective is to develop and optimize aquacul- with water sterilized by UV and changed every two ture techniques of three species of clams having days; larvae fed daily on a mixed diet of Iso, Mn, Ts commercial interest: the carpet shell Ruditapes and Cht. Water temperature varied for each species decussatus), pullet carpet shell (Venerupis pullastra) : between 22 and 24ºC for the short necked clam and short necked clam (Tapes philippinarum). and between 18 and 21ºC for the two other species. The success in the procurement of spat depended on In 2005 in the autonomous community of Galicia, the species. The carpet shell clam exhibited differen- the following experiments were carried out in kee- ces in the clutches, and in some cases, the larvae ping with the program to carry out the different died just a few days after being cultured. At other lines of action: times spat was obtained, but in all cases the percen- tage of survival was very low from larval stage D to Hatchery conditioning. The conditioning of the 500µm sized spat. Pullet carpet shell clam: larvae carpet shell clam was carried out in two trials –one were cultured from parent stock taken from using parent stock taken from Camariñas in January Puentedeume. The parent stock was brought to the and another from Villaviciosa (Asturias) in March. hatchery in spring, with spawning taking place The experiments were conducted according to the immediately after their arrival. The larval culture did protocol laid down by the participating autonomous not present any major problems and the survival communities: temperature 19ºC ± 1º and diet consis- rate (from larval stage D to 500µm sized spat) raged ting of a mixture of six species of microalgae: Tiso, from 30-40%. As far as the short necked clam is con- Mn, Ts, Cht, Sk y Pht, administered daily on a conti- cerned, the culture consisted of larvae taken from nuous basis. The amount administered was 6% of mature parent stock collected from the natural envi- the dry weight of parent stock; water renewal: 30 l/h ronment on the Isla de Arosa in summer. In general and kg. The parent stock used in both tests were in this culture did not suffer high mortalities. However the early stage of gametogenesis. The duration of it was found that low temperatures tended to slow the conditioning process was the same for both and down the growth rate of the larvae and postlarvae. after one month and a half, all the clams were fully mature and after two months they spawned.

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Grow-out. One of the methods proposed in this results, it is possible to conclude that there is a clear project is directed at using part of the particulate seasonality in growth, limited by water temperature organic matter found in the effluents produced by conditions and that the growth potential is higher turbot fish farms. The growth of hatchery-reared in the pullet carpet shell clam, followed by the short spat of several species of mollusks was studied: the necked clam and trailing far behind, the carpet shell carpet shell clam (Ruditapes decussatus), the short clam. The mortality recorded correlates directly with necked clam (Tapes philippinarum) and the pullet the initial spat quality (a very important factor in the carpet shell clam (Venerupis pullastra). Two criteria success of this culture stage) and the type of metho- in the handling of the spat were tested: the thinning dology used. The system using bags suspended from out and sorting processes as warranted by the den- rafts was found to be highly suitable for grow-out sity or size of the individuals, and on the basis of dif- owing to the low mortality rates recorded and, par- ferent initial biomass values per unit of culture (3, 6 ticularly, because of the way in which it uses the and 18 kg/m2), keeping the density of each batch of available water, leading to very high with growth spat stable until the specimens reached an average rates which make it possible to reduce grow-out length of 10 mm. The spat were reared in PVC drums times and increase the cost-effectiveness of the faci- 32 mm in diameter with a 1-2 mm mesh, depending lities. on the size of the spat, suspended in pools. The top part of the drums received the flow of effluents Pathology. We examined the parasites and patho- from the fish farm. A inverted forced flow system of logies and carried out a bacteriological study on the roughly 12 L/min was used for the circulation of the three clam species in the different grow-out tests water in the drums. Parallel to this experiment, and on the parent stock used in the hatchery to pro- other tests were conducted using raw seawater, cure spat. When the spat reached the desired size, they were shipped to the fishermen’s associations to be sown For the pathological study, three histological inci- in the culture areas. sions 5µm thick were made on 25-30 clams of each species, using both spat and adult specimens. The In these types of installations it is possible to ripen results show that the parasites found in the clams bivalve mollusk spat all year round. V. pullastra is would not cause any pathological harm, owing to the species that was found to best adapt to the their low prevalence and intensity. Of all the patho- autumn-winter conditions. As the temperature logies observed, it is the presence of rickettsias in increases, the most suitable species for grow-out is T. the digestive tubes of the carpet shell clam that philippinarum, while R. decussatus exhibited irregu- would potentially present the greatest degree of lar growth throughout the year. Spat with a weight pathogenicity. In any event, the low levels of inten- of 3 mg and over were found to grow even in win- sity and prevalence in spat seems to indicate that it ter conditions. would not be harmful. Perkinsus atlanticus was not observed in any of the clams in the grow-out pro- Other experiments in the area of grow-out were cess. At the start of grow-out, only ciliates were carried out using the raised bag method in the inter- observed in the gills and digestive system, with the tidal zones of the beaches of O Grove (Illa Toxa) and number of parasites and commensals having increa- Vilaxoán (Borreiros) as well as the modular bag sed by the end of the experiment. system suspended from rafts in O Galiñeiro park, -all of the above located in the Ría de Arousa. In these In the parent stock of the carpet shell clam from tests we used Size 3 spat from the three clam species Villaviciosa (Asturias) it is important to highlight the having wet weights (PHind) ranging from 10-80 mg, presence of trematodes, but with a low prevalence holding them at different densities and subjected to and no signs of any major pathologies. Perkinsus a number of different types of monitoring: oceano- was not observed. In the pullet carpet shell clam graphic conditions, demography, biochemical com- from Puentedeume the presence of Perkinsus was position, histology and bacteriology . The start and detected, -13.3% of the clams were seriously infec- duration of each test were as follows: ted. Trematodes were also observed, but owing to their low prevalence, were not considered impor- The grow-out stage ended when the clams rea- tant. In the short necked clam from the Isla de Arosa ched sizes ranging from 12-17 mm and live weights Perkinsus was not present and in general this clam of between 200 and 900 mg. On the basis of the exhibited very few parasites.

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The bacteriological study consisted of taking a be considered healthy in comparison to references count of aerobic organisms using Marine Agar pla- of diseased individuals presenting higher counts. In tes and vibrions with TCBS. A homogenized prepa- any case, we believe that the species must be identi- ration was made using the flesh of 15 adult clams fied in order to determine –in the event of mortali- and 12 gr. in the case of spat. The results showed ties- the relationship of pathogenic species with a than none of the bivalves were free of bacteria and high bacterial load. that most of these marine bacteria are not harmful to adults, which are able to tolerate higher concen- trations than the spat. The clams in this study may

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Promotion of the culture of new sparid species: Linares Cuerpo, F. Sea bream. Pilot trials and technological transfer. Coordinated project

START: January, 2004 RESEARCHERS: FINISH: December, 2007 IEO de Vigo: Olmedo Herrero, M.; Peleteiro Alonso, J.B. IGAFA: Rodríguez Villanueva, J. L. FINANCIAL ASSISTANCE: DXRM: Rodríguez González, L.M. Plan Nacional de Cultivos Marinos. Junta Nacional Asesora de Cultivos Marinos. RESEARCH SUPPORT: (National Program for Marine Culture – Marine CIMA: Cores González, M. J. Culture Advisory Board) CETMAR: Pazos Sieira, G. (JACUMAR) IEO: Álvarez-Blázquez Fernández, B; Gómez Ceruelo, C.

INTERNS: Rodríguez Díaz, R; Soto Fraga, N.; Cachafeiro Diéguez, P.

SUMMARY This study is part of a nationwide project with the weights of 26.46±7.52 g and 9.94±1.01 cm. The re- participation of 8 autonomous communities. The sults showed no significant differences in the purpose of this project is to promote and transmit growth of the fishes administered the two types of the technology for the culture of new sparid species: feed, with the animals having weights of white sea bream (Diplodus puntazzo), couch’s sea 190.07±28.23 and 195.42±32.12 g and sizes of bream (Pagrus pagrus), bluespotted sea bream 17.95±1.50 and 17.96±1.47 cm at the end of the (Pagrus auriga), gilt head sea bream (Dentex den- experiment for fishes given feeds of 12% and 7% tex), and the blackspot sea bream (Pagellus bogar- respectively. aveo) to be used by different companies. As regards the biochemical composition of the The main objective of this project in Galicia is to blackspot sea bream fry, the lipid content of the develop a suitable type of feed to meet the nutri- fishes was found to increase in the liver over the tional requirements of the blackspot sea bream in course of the experiment rising from values of 23% order to optimize its growth and reduce fat content. to 35% of the dry weight, while the lipids in the The specimens used in the different experiments muscle were in the range of 11-13% of the dry were reared at the CO of the Oceanographic weight. No significant differences were found in the Institute of Vigo and the experiments carried out on total lipid content of the liver and muscle of the two rafts adapted for the culture of fishes- one blackspot sea bream fry fed the two types of feeds. belonging to the Association “Illa do Santo” of Bueu However there was a slightly higher protein content (Raft 1) and the other, belonging to the Local in the liver of specimens given the feed 50/12 (36% Ministry for Fisheries and Maritime Affairs (Raft 2), of the dry weight) than in those fed with the 50/7 both located in the Ría de Pontevedra. (28% of the dry weight). In terms of fatty acids, the total PUFA values were higher in the livers of fry fed Raft 1. Experiment I. The purpose of this experi- with having 7% fat, with the total contents ment is to compare the effect on the growth and accounting for 44% of the total fatty acids versus body composition of blackspot sea bream fry of the 38% in those given a diet having 12% fat.. The 22:6 use of two types of feed manufactured by the com- (n-3) accounts for 19% of the total fatty acids in the pany SORGAL containing a different composition in liver of the fry fed with the 50/7 feed as compared protein/lipids, 50/12 and 50/7. The experiment, pro- to 16% in fry fed with the 50/12 feed. grammed to last one year, was started in April, 2004 with 7000 specimens having mean initial sizes and

42 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

Experiment II. A comparison of the growth, body Raft 2. This experiment compares growth and composition and coloration of the fishes from the body composition of juveniles fed with three types previous experiment (Pm 192.75±30.18 g and Tm of feed. The feeds were designed on the basis of the 17.96±1.47) fed with two types of feed –one having hypothesis that the use of feed having a fat content a composition of 50/18 and supplemented with of less than 16% in the diet of animals in the fatten- carotenoids (B-carot) and the other, SORGAL 50/12 ing stage has no clear effect on the composition of (S-12) used as a control. In June, de 2005 5 batches the specimens. Based on this hypothesis, the % of of 900 fishes were prepared. Three of these batches fat in these feeds is partially replaced by carbohy- were fed B-carot, while the other two were given S- drates from different raw materials. 12. After four months on these diets, no differences in growth were found between the two groups, The experiment was carried out in duplicate using with the mean values being Mw 319.05±50.62 g and 6 batches of 1125 juveniles all taken from 2004 Ms 20.75±1.38 and Mw 317.45±39.43 g and Ms Tm spawnings. In June, 2005 they were placed on the 20.93±1.15 cm for fishes fed with B-carot and S-12, raft in six modules: P1 50/4 (mean weight respectively. 118.97±3.86 g and mean size 15.70±0.18 cm), P2 52/9 (mean weight 102.06±0.48 g and mean size Two other batches of 500 fishes hatched in 2001 14.95±0.24 cm) y P3 64/7 (Mean weight 92.52±8.09 g (Mw 664.55±111.12 g and Ms 25.73±2.17) were and mean size 14.66±0.54 cm). introduced into two other modules on the same date, and one batch fed with B-carot and the other After 6 months, the following mean values were with S-12. In October, 2005 the fishes fed with the attained: P1, mean weight 229.6±25.86 g and mean two types of feed did not exhibit any differences in size 19.34±0.72 cm, P2 mean weight 206.62±25.82 g growth and reached a mean weight of and mean size 18.59±0.89 cm and P3 mean weight 801.62±142.01 g and 789.39±125.50 g and a mean 207.10±34.02 g and mean size 18.56±1.09 cm). size of 27.82±1.78 cm and 27.68±1.85 cm - fishes fed with B-carot and S-12 respectively. The perivis- The specific growth rates during this period were: ceral fat content of the fishes was 4% of the body 0.31, 0.33 and 0.37 for the groups fed P1, P2 and P3 weight for the two groups of fishes. respectively and the condition factor was 0.03 for all the groups during the entire period. This experiment was carried out as a master’s thesis in Aquaculture on the Canary Islands and is This experiment will continue throughout 2006, discussed in greater detail in section 3.2 of this during which time, in addition to monitoring the annual report. growth of the specimens, we will also carry out bio- chemical analyses.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Techniques for the minimization, treatment and Guerrero Valero, S. use of aquaculture residuals. RESEARCHERS: Subproject: A study of the evaluation of fish CIMA: Alonso Fernández de Landa, J.L. byproducts in fish farms. Biochemical and microbi- USC: Estévez Toranzo, A.; Pascual López, C.; ological aspects. López Romalde, J.; Barja Pérez, J.L.; Rodríguez Vázquez, C.J. START: January, 2005 FINISH: December, 2007 INTERN: Otero Otero, M. FINANCIAL ASSISTANCE: Plan Nacional de Cultivos Marinos. (National Program for Marine Culture (JACUMAR)

SUMMARY This project, developed in collaboration with the of different species, such as man, for example. Here autonomous communities of the Canary Islands, it is important to stress that the use of certain Cataluña and Andalucía and coordinated by AZTI, “desirable” bacterial species during the fermenta- aims to provide the Spanish aquaculture sector with tion process which are resistant to the processing real solutions that are effective and economically conditions, may release substances (biocines?) into feasible to minimize and reduce in origin the waste the fodder that control or inhibit other bacterial generated from this activity. It also seeks to obtain species, of special interest in pathogens, as occurs compounds of a high added value for commercial during the fermentation of some food products apt application and to facilitate the use of the remain- for human consumption (cheese, for example). ing residuals. This will contribute to the develop- These starter cultures are, therefore, an important ment of the sector and to the attainment of sustain- channel in the ensilage process for aquaculture pro- ability, in other words, to obtain a greater yield ducers. from the raw materials and reduce the amount of waste, residuals and gas generated, making better Objectives use of the available resources, while, at the same time, respecting the environment. All of this is to be To gain knowledge of the true situation of the carried out in a way that is economically cost-effec- generation of residuals produced by aquacul- tive and in compliance with the current environ- ture in Galicia. mental regulations, which will improve the compet- Find out the volumes and classification of the itiveness of the sector. Today, given the enormous residuals generated, as well as their current development of aquaculture in Europe, highlighting management. Spain as the most important producer, the aquacul- Facilitate the treatment and updating of data ture sector is faced with the problem of the genera- by means of a unified and updated database. tion of waste from the production and processing Find out the legal requirements and condition- involved in this activity. These residuals are general- ing factors applicable to each type of residue ly used to make fish meal or they are burned, which produced by aquaculture in Galicia and their is a costly process. However, these residuals, which possible use. are, in principle, innocuous, may be used for nutri- Identify the most efficient and cost-effective tional purposes by these species or others that are to means to reduce the volume of residuals gener- be cultured, in such a way that they may be carriers ated in origin through innovative minimization of intraspecific pathogens, as happened in the case techniques. of mad cow disease, which may have repercussions Identify useable elements in aquaculture on other populations reared in other aquaculture byproducts and develop systems for procure- farms (Gill, T., 2000), or they may even be pathogens ment/extraction.

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Develop an Innovation Plan for aquaculture process took 72 hours to attain the appropriate vis- byproducts. cosity. The microbiological ensilage was started by Design protocols and innovative technologies using Lactobacillus plantarum as an inoculum and to reuse aquaculture byproducts and obtain dehydrated milk whey. The process took 7 days to high value products. Determine the feasibility attain a consistency similar to that in the chemical of obtaining such products on an industrial ensilage process. All of the process were carried out level. at 25ºC. Propose real and effective management alter- natives for non-useable residuals in compliance The results of the biochemical analyses demon- with the current legislation. strated that during the ensilage process and subse- Facilitate the dissemination and good use of the quent storage, the moisture, chlorine and ash con- results and conclusions of the project aimed at the tent remained at constant levels. The protein frac- Spanish aquaculture sector through reports pub- tion, however, underwent drastic variations. In con- lished in guide and leaflet format. trast to the studies that have already been pub- lished, here the loss of protein content was high. Development and current state of the project The percentage of total nitrogen remained con- stant. However, in the case of the ensilage of turbot, The generation of biological residuals is one of after the first day of the chemical process, roughly the major problems facing the aquaculture industry 50% of the total nitrogen total was not proteic owing to the high cost incurred in complying with nitrogen and it reached 70% after 30 days. The same the EU Directive on environmental protection regu- occurred with the microbiological ensilage after 70 lations. In Galicia, the most important production is days. In the case of trout viscera, the protein fraction the rearing of turbot in marine hatcheries and trout was very low, and the level of non-proteic nitrogen in freshwater fish farms. A possible solution to this was 70% after one week of ensilage. The lipid frac- problem is the ensilage of organic waste (dead ani- tion was very high in comparison with the turbot mals and viscera) from farms to minimize costs, ensilage, and required extraction. reduce risk material and provide feed for other ani- mal species. In general, the largest fraction of nitrogen is non- proteic, although there were small peptides, amino In this paper we present the preliminary results acids and amines. We found no evidence of any with to types of ensilage – chemical and microbio- deterioration or alteration of the ensiled products logical using two types of residuals – dead turbot during storage in terms of the formation of total and trout viscera. In the chemical ensilage, we used volatile nitrogen. formic acid (3%) and potassium sorbate (0.1 %). The

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PROYECT STAFF

TÍTLE CHIEF RESEARCHER: Sustainable Harvesting of Ensis (SHARE-90). Niall McDonough (Ireland)

START: March, 2004 CHIEF RESEARCHER AT THE CIMA: FINISH: March, 2007 Martínez Patiño, D

FINANCIAL ASSISTANCE: RESEARCHERS: Trans-national Partnership Agreement INTERREG CIMA: Da Costa González, F; Nóvoa Vázquez, S; IIIB “Atlantic Area” Ojea Martínez, J.

RESEARCH SUPPORT: Álvarez Llamas, L.M.; Bouzamayor Yáñez, M. V.; Cotarelo Jardón, M. J.; Páez Pérez, F.J.; Ruíz Misioné, M.

SUMMARY Progress has been made in the different actions in The larval culture was carried out using the meth- the development of technologies for the culture of ods developed and refined in our hatchery. A test solenaceans of the genus Ensis. was conducted on E. arcuatus to determine the opti- mum temperature for the development of larval The trials carried out for the conditioning of the culture, with temperatures of 14, 18 and 24º C. The razor clam (Ensis arcuatus) under 2 temperature temperature at which optimum growth was regimes have demonstrated that temperature is not achieved was 18º C, while the best survival rate was a factor that affects the conditioning of this species. obtained at 14º C, although the latter larvae took A temperature gradient ranging from 20 ºC and the the longest time to attach (26 days). temperature of the natural environment was used. The animals reached maturity earlier than in the We also carried out experiments with different natural environment in the two cases, but no signif- diets consisting of a mixture of microalgae in razor icant differences were found between the groups. clam larvae: diet 1 (Isochrysis galbana+Paulova This made it possible to use the natural environment lutheri); diet 2 (I. galbana+P.lutheri+Chaetoceros temperature, thereby reducing costs, as the water calcitrans);diet 3 (I.galbana+C.calcitrans+ Tetra- did not require heating. selmis suecica) and diet 4 (T. suecica+I. galbana+P. lutheri+C. calcitrans). After being fed the different One aspect that might limit the production of diets, the larvae exhibited a similar increase in hatchery-reared bivalve larvae is the lack of suit- length. The only differences were observed in sur- able methods available to induce spawning. In the vival, with the best survival rate being obtained two species of Ensis se two methods of induction using diet 4. were successful after carrying out trials with differ- ent stimuli. In Ensis siliqua thermal shock induction Different systems for the maintenance of the post- proved to be effective, while in E. arcuatus the larvae and spat were analyzed to find out which one most effective process was carried out without would produce the best yield in the growth and sur- making any changes in the water temperature, vival of the postlarvae. This was carried out as fol- i.e., keeping it the same as in the natural environ- lows: Two containers with different areas 0.125 and ment and changing the level (column) of water, 0.5 square meters were used with and without sub- including brief drying out periods, simulating the strate. The postlarvae were held in these containers effect of the tides. in 150 L larva culture tanks. Once the larvae reached 10 mm, they were transferred over to two different system for 2 weeks. Both were kept in sieves with

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sand and an inverted water flow, with the tank vol- The controlled containers consisted of cages made ume ranging from 150 L to 5000 L. out of steel rods 50 cm long with a 2 mm mesh. The razor clams were sown at a density of 200 individu- At the beginning of December, 2005 the razor als /m2 and a size of 39.09±4.03 mm. Every two clam (E. siliqua) was sown under controlled condi- months the containers are monitored and one of tions in the intertidal zone of the Ría de Ribadeo. the cages sampled.

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Optimization of settlement processes in bivalve lar- Guerra Díaz, A. vae reared in mini-hatcheries. An analysis of spat yield with different food sources. RESEARCHERS: CIMA: Martinez Patiño, D.; Nóvoa Vázquez, S. START: July 2004 IGAFA: Andrés Rivas, M.C.; Figueiro Casas, R.; FINISH: July, 2006 Lastres Couto, M.; Martínez Rubal, A.

FINANCIAL ASSISTANCE: RESEARCH SUPPORT: Plan Galego de Investigación e Desenvolvemento Búa Santamaría, I.; Loureiro Barcón, J.M.; Tecnolóxico - (Galician Program for Research and Miranda Bamio, M. Technological Development) (PGIDT) PGIDTCIMA 04/4

ABSTRACT The objective of this project is to promote the effi- The hatchery stage should be aimed at producing cient management of the facilities of the mini- massive quantities of small spat (< 2mm) to be hatcheries located in O Vicedo (Lugo) and Camariñas “grown-out” either in other facilities or outdoors. (A Coruña) in keeping with the aspects outlined in For this process to be carried out with a certain the title of this action. For this purpose experiments degree of regularity and efficiency, it is necessary to have been designed and carried out at the facilities have large qualities of seawater available with the of the IGAFA, the results of which have been pro- correct amount of food, adjusted to the filtration jected in the units as outlined. capacity of the spat. In keeping with this, and sup- plemented with other projects, different grow-out The tests to improve efficiency in the metamor- systems are evaluated with different food sources: phosis stage in which the natant larvae become ses- natural (on the raft), with a production of a natural sile, were based on reducing the stress caused by bloom in large outdoor pools with water coming changes and normal handling processes during the from effluents from a seawater fish farm located on larval stage. During metamorphosis, the contribu- land. A number of different factors come into play tion of microalgae was reduced to less than 50%, in all of these systems: minimum size at which grow- and we made sure that the doses of microalgae con- out may be started, recapture rates and growth sisted of a high proportion of diatom cells. In the lar- depending on the species, installation and handling val culture of the different clam species, it is neces- costs. All of these factors together will determine sary to take into account a number of considerations the efficiency of the system. The size ranging from to improve attachment efficiency. The first is to start 1.5–3 mm should be held at land-based facilities in attachment with larvae in the “mature” stage of the containers with an inverted forced flow system and metamorphosis process –larvae that do not pass the food source being a natural bloom. After this through a 210 µm sieve. A minimum amount of han- size, (4 mm, T2), the spat may go on to be reared dling, constant temperature and appropriate diet in any of the aquaculture systems currently being are determining factors to improve efficiency in tested: in rigid mesh bags, in drums with inverted metamorphosis. Also required is the correct selec- flow from effluents –with large-particle filtration- tion of larvae to obtain a pool with a constant and from marine fish farms; spat surrounded by the cir- homogeneous growth that will ensure that larvae culation of water from zones of high natural pro- are robust during the evolution of the process of ductivity, etc. Other determining factors to guaran- metamorphosis. tee efficiency and cost-effectiveness in the grow-out stage are a feasible investment, a suitable location, and simple, routine handling of the spat.

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PROJECT STAFF

TITLE: TECHNICAL COORDINATION: An initiative to culture the Pacific oyster (Crassotrea Rodríguez González, L. (DXRM) gigas) in different Galician Rías. ADMINISTRATIVE COORD.: START: May, 2004 Fernández Cañamero, M.L. (CETMAR) FINISH: December, 2006 RESEARCHERS: FINANCIAL ASSISTANCE: CIMA: Guerra Díaz, A.; Montes Pérez, J. Dirección Xeral de Recursos Mariños (DXRM). INTECMAR: Fernández Conchas, R.D.; Pérez Bello, J.L. Consellería de Pesca e Asuntos Marítimos. IMIDA: García García, J.; García García, B. (Directorate General of Marine Resources. Local CETMAR: Iglesias Estepa, D. Ministry for Fisheries and Maritime Affairs) Xunta de Galicia. RESEARCH SUPPORT: Meléndez Ramos, M.I.; Penas Pampín, E.

SUMMARY The activity of the Galician oyster farming sector is mercial size (category 3 on the French market: 75 g), based primarily on the culture of the European flat in most of the points studied, after 14 months of cul- oyster Ostrea edulis. Due to the fact that this species ture. The values of accumulated mortality at the end has a high mortality rate and that the availability of of the culture period did not generally exceed 15%. spat varies greatly from year to year, it was decided The condition of the oysters generally presented a to attempt to culture the Pacific oyster Crassostrea variation pattern that could be directly linked to gigas to supplement the culture of the European their gonad maturation cycle, so that the lowest val- flat oyster. The Pacific oyster has higher growth ues were recorded towards the end of autumn- rates and lower mortalities. However, since it is an beginning of winter, rising sharply in spring which allochthonous species, some doubts have been coincided with the increase in water temperature raised as to its possible environmental impact on the and reaching maximum values in summer. At the Galician Rías. Although the first experimental cul- end of the summer we detected a decrease in condi- tures of the Pacific oyster were authorized in 1991, tion index values, which was relatively pronounced the development of this activity was not considered depending on the culture zones, coinciding with the until now. This project, promoted by the Directorate release of gametes. Towards the end of autumn, General of Marine Resources, aims to assess produc- there was a slight increase in the condition of some tive, environmental and technical aspects related to of the batches studied. As regards the existence of the feasibility of the culture of C. gigas in the malformations, the only condition of this type Galician Rías. A study is being carried out at the observed was the presence of gel chambers on the CIMA on all the biological aspects affecting the eco- inner side of the valves in a large percentage of the nomic profitability of the culture of this species oysters cultured in park Redondela A, which would (growth, mortality condition indices, resistance to probably affect their being accepted in the commer- drying out, malformations) and other critical aspects cial market. fundamental to the culture of foreign species (asso- ciated pathologies and reproductive potential). To With regard to the study of the possible risks asso- this end the experimental culture of a series of ciated with the culture of Crassostrea gigas, the batches of the Pacific oyster imported from monitoring of the pathology by means of the analy- Arcachon (France) is being tested on rafts with an sis of histological incisions did not reveal the pres- experimental concession for this species, in addition ence of parasite organisms or diseases that may pos- to an intertidal concession from the fishermen’s sibly be responsible for mortalities of bivalve mol- associations of Vilaxoán and Ribadeo. lusks. The evaluation of the reproductive capacity of C. gigas in the Galician Rías was approached by The growth rate was very high as demonstrated means of a study of the gametogenic cycle of adult by the fact that the spat sown in May, 2004 having a oysters reared on rafts and in the intertidal zone, as mean weight of 1.48 (± 0.56 D.E.) g reached com well as by placing commercial collectors in the dif-

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ferent areas where this animal has undergone reached in June-July, and spawning taking place experimental culture in order to determine if any between July and August. Gonad reabsorption Pacific oyster spat was procured. Also analyzed were occurred in late summer. A preliminary analysis of the the time series of records of different oceanographic collectors showed that no spat were procured at the variables corresponding to the sampling stations of points located in the Ría de Arousa, but some spat the monitoring network of INTECMAR in the last 6 were obtained from the collectors located in the Rías years, to determine if at any point in time the condi- of Ribadeo and O Barqueiro. This observation agrees tions were suitable for the larval survival of C. gigas with the analysis in the oceanographic records, since and, consequently, the existence of recruitments of the rías in the north of Galicia exhibited suitable tem- this species in the Galician Rías. The evolution of the perature and salinity conditions all year round so that gametogenic cycle was similar to that of the oysters the larval development of the Pacific oyster would be reared on rafts and in the intertidal zone, with the carried out successfully. gonad developing during spring, maturity being

50 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Restocking of turbot on the Costa da Morte Linares Cuerpo, F. (NW Spain). RESEARCHERS: START: February, 2006 IGAFA: Rodríguez Villanueva, J.L.; Mariño, C. FINISH: December, 2006 RESEARCH SUPPORT: FINANCIAL ASSISTANCE: Cid González, E.; Fernández Franco, B Dirección Xeral de Innovación e Desenvolvemento Pesqueiro. Consellería de Pesca e Asuntos Marí- timos. (Directorate General of Fisheries Innovation and Development. Local Ministry for Fisheries and Maritime Affairs) Xunta de Galicia.

SUMMARY This research action has led, for the first time, to From April to September 2005 fishes earmarked the implementation of a restocking program whose for restocking were obtained. Different companies main objective is to recover the stocks of several were contacted - Pescanova, Stolt Sea Farm and the species on the Galician coast. group Isidro de la Cal. Participating in the program was Dr. Paulino Martínez (Genetics Department of Before devising the restocking plan of a species, the School of Veterinary Medicine of Lugo), as advi- the species must fulfill certain requirements that will sor of the genetic monitoring of the fishes, and the indicate whether or not the program has a chance diversity of the organisms to be restocked was to succeed. The characteristics are as follows: favored as much as possible. Ten thousand turbot juveniles weighing between 10-20 gram were pur- 1. The species must be autochthonous. chased from two of the above companies and the 2. The commercial catches of the species must fishes were taken to the Galician Institute of have declined. Aquaculture Training (IGAFA). During the months of 3. Its market price must be high. June, July and August, the fishes were fattened and 4. It must grow rapidly. acclimated until they attained a weight of 30-40 The culture technology must be widely known. grams. During this period, routine maintenance was First catch weight must be high (to improve the carried out – feeding, cleaning, measuring parame- profitability per cost of unit produced). ters (O2, Tª, salinity), samplings and other monitor- ing tasks. The fishes were later tagged with T-anchor The objectives of this action are outlined as fol- tags whose base is T-shaped and therefore remains lows: anchored in the body of the turbot. These tags bear a number for each individual and the telephone 1.- To determine the survival, growth and distribu- number of IGAFA. In this way, if the fish is caught, tion of 10,000 turbot fry (with a mean weight of 30 the fisherman will be able to contact the people grams) during the months after their release to involved in the program. three beaches with different types of hydrodynam- ics: a beach exposed to the open sea (Playa de During the period that the tagged fish remained Nemiña), a semi-exposed beach (Playa de Moreiras) in the tank, no injuries or mortalities were detected and a beach located in the inner ría of Camariñas owing to tagging and the percentage of tags lost (Playa de Lago). was very low. Also during this period, the organisms 2.- Find out if the tagging techniques and tags underwent microbiological monitoring through the used are effective in identifying individuals at the collaboration of Dr. J. Luis Barja (Microbiology time of recapture. Department of the University of Santiago).

3.- Ascertain the reaction of the sector to restock- For the purpose of determining the effect of the ing plans. adaptation of the fishes to the marine environment,

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prior to release, to facilitate their acclimation to the The fishes were released by means of nets and natural environment, 15% of the fry were held for their reaction was to swim to the bottom in a mat- a period of (15-20 days) in a 25 m2 pool with a sand ter of seconds. bottom, where they were fed live prey, preferably amphipods and mysidaceans. No moralities were The fishes that were released have been charac- observed when these animals were fished from the terized as to parent stock and they also underwent tank, although some tags had to be replaced. a microbiological control before being released. A number of different samplings were carried out In September a meeting was held with the skip- in the release areas in November and December. pers of the fishermen’s associations located in the release area from Muros to Caión to provide them The released turbot will be monitored by sam- with a detailed explanation of the restocking plan pling which will serve to estimate the survival rate, and to ask for their collaboration. growth and distribution. This monitoring will be included in a broader restocking plan that will On October 8th, the fishes were moved to the involve new releases of turbot and will also be release areas. The animals were placed in tanks extended to other species, such as the lobster. where the temperature and oxygen were monitored Moreover, parallel to this, a campaign will be carried and transported by truck. These tanks were loaded out to increase the awareness of the fishing sector. onto a vessel to be taken to the selected beaches.

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3.1.2. AREA OF PATHOLOGY

PROJECT STAFF

TITLE: CHIEF RESEARCHER: A study of disseminated neoplasia and the patho- Carballal Durán, M.J. logical alteration known as “large foci with a strong hemocytic reaction” affecting the cockle, RESEARCHERS: Cerastoderma edule, of Galicia. CIMA: Villalba García, A. CETMAR: Iglesias Estepa, D. START: September, 2002 USC: López Romalde, J. FINISH: September, 2005 IGAFA: Lastres Couto, M.

FINANCIAL ASSISTANCE: RESEARCH SUPPORT: Plan Galego de Investigación e Desenvolvemento Meléndez Ramos, I.; Penas Pampin, E. Tecnolóxico - (Galician Program for Research and Technological Development) (PGIDT) INTERNS: PGIDT02RMA50101PR Angueira Souto, M.C.; Díaz Costa, S.M.

SUMMARY The populations of the cockle, Cerastoderma study. In 2005, a decrease in the prevalence of this edule, are dominant in many bivalve beds in Galicia disease was observed. and they have an important commercial value. These populations undergo episodes of mortality The FGIHI alteration was only detected in cockles associated with the drops in salinity caused by flood- from Sarrido, with a much lower prevalence than in ing. However, our studies in recent years point to Lombas del Ulla. The prevalence values for FGIHI the presence of two serious pathologies also associ- varied from month to month without presenting a ated with a high mortality rate: disseminated neo- seasonal pattern. The prevalence of FGIHI was plasia and the disease known as “large foci with a greater in 2004 than in 2005, decreasing in 2005. strong hemocytic reaction (FGIHI)” This project was started in 2002 to continue the studies on these two The temporal variation of temperature and the diseases and to assess their incidence in the cockle salinity of the seawater were compared with the populations and to determine their origin. temporal variation of the prevalence of FGIHI and neoplasia. To do this, we used data recorded week- Activities carried out. The temporal dynamics of ly on the temperature and salinity in the beds under the prevalence and intensity of disseminated neo- study and measured by INTECMAR and data on plasia and FGIHI was examined. To do this, we col- these variables measured when the monthly sam- lected monthly samplings of 25 commercial size plings were taken. No correlation was found cockles from the natural beds of Noia, Sarrido between these variables and the monthly preva- (Cambados) and Lombos de Ulla. They were then lence of FGIHI. In the case of neoplasia diseminada, subjected to a histopathological examination. In the no correlation was observed between the temporal three beds under study, we detected variations in variation of this disease and the salinity and temper- the prevalence and intensity of neoplasia. In Noia ature of the seawater. However, the peaks of maxi- the bed appeared to follow a seasonal pattern, with mum prevalence of neoplasia were correlated with a decrease in prevalence in late summer and an drops in temperature. increase at the beginning of spring and autumn. In Sarrido in 2004, there was a trend similar to that of Studies on the ultrastructures of tissues affected Noia. However in 2003, this was not the case and the by FGIHI disease have demonstrated the presence of highest values were observed in August. The viral particles in the cytoplasm of cells that are the Lombas del Ulla area did not exhibit a seasonal pat- foci of the inflammation. These particles have char- tern. The mean prevalence values of neoplasia were acteristics resembling the picornaviruses. The cells greater in 2004 than in 2003 in all the beds under with the virus were extremely degraded, for which

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reason it was not possible to determine if these cells demonstrate the inverse transcriptasa activity in were hemocytes or the remains of an unidentified these cells. The results of these studies confirm the protist parasite. Studies are underway to identify activity of this enzyme and its absence in the these cells. hemolymph cells and tissues of non-neoplasic cock- les. This would confirm the viral origin of neoplasia. The study of the ultrastructure of these cells in the hemolymph has shown that they are larger in size To determine the presence of disseminated neo- than the hemocytes, exhibiting a large pleomorphic plasia in other bivalves, samples of clams were taken nucleus, and a very pronounced nucleolus and a from the beds under study. We did not observe the cytoplasm, with very few organelles. The ultrastruc- presence of neoplasia in the clams Venerupis pullas- tural characteristics of the neoplasic cells observed tra, Ruditapes decussatus or Ruditapes phillip- in the tissues showed similar characteristics to the inarum. neoplasic cells in the hemolymph. Flow cytometry techniques have shown that neoplasic cells have dif- Trials were carried out to study the possible trans- ferent levels of ploidy: hypodiploids, hyperdiploids, mission of neoplasia between cockles and neoplasic triploids tetraploids, pentaploids, sexaploids, hepta- cockles to Ruditapes decussatus. Neoplasia was ploids and octoploids. The ploidy level was found to found to be transmitted from cockle to cockle increase with the intensity of the neoplasia. through the cohabitation of neoplasic cockles and those that are free from the disease in open-circuit Through ultrastructurale studies, we observed the tanks and by injection of neoplasic cells to cockles presence of viral particles similar to a retrovirus in not affected by this disease, The results of the exper- some neoplasic cells. To confirm this possible viral iment on the transmission of neoplasia from cockles origin of the disease, trials were carried out to to clams are being analyzed.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Commercial solenaceans in Galicia after the López Gómez, C. Prestige catastrophe: the impact of the oil spill on some biological aspects. RESEARCHERS: CIMA: Da Costa González, F.; Guerra Díaz, A. START: November, 2003 CETMAR: Darriba Couñago, S. FINISH: November, 2005 Delegación de la CPAM A Coruña: Catoira Gómez, J. L. INTECMAR: Álvarez Álvarez, C. FINANCIAL ASSISTANCE: Plan Galego de Investigación e Desenvolvemento RESEARCH SUPPORT: Tecnolóxico - (Galician Program for Research and Meléndez Ramos, I.; Miranda Bamio, M.; Technological Development) (PGIDT) Penas Pampin, E. PGIDIT03RMA50101PR

SUMMARY The objective of this project is to study the effects the presence of other parasites (Ciliates, copepods, of the catastrophe of the Prestige on commercial tremetods sporocysts and infiltrations) that show no populations of solenaceans, the razor clams (Ensis clear pattern. arcuatus) and (Ensis siliqua). The effects are studied on a pathological level (histopathology and bacteri- Bacteriological study. A count of the total bacte- ology), immunological level (hemogram variability, ria was taken on Marine Agar (MA) plates and in the hemocyte viability, enzymatic ability, etc.), reproduc- selective medium for Vibrios (TCBS). In contrast to tive cycle and larval development. Samples of Ensis the first year (2204), in Ensis arcuatus the highest arcuatus were taken in San Martiño-Cies (an affect- total bacteria count was observed in the Cíes (affect- ed area) and in Means-Cambados (an unaffected ed), although with slight differences. During the sec- area); and samples of Ensis siliqua were collected in ond year the total count of Vibrios was higher in Langosteira–Fisterra (an affected area) and in Barra- Cambados (not affected), also with small differ- Cangas (an unaffected area). ences. In any event, the higher presence of Vibrios in Cambados may be attributed more to its location Histopathological study. The data obtained from (inner ría zone) near the effluents from rivers and the razor clam beds located in San Martiño-Cies urban areas that to the effect of the Prestige. The (affected) and Means-Cambados (not affected) total bacteria count and Vibrios count for Ensis sili- demonstrate that certain pathologies only appear in qua were higher in Cangas (not affected). The dif- the Cíes bed (gregarines, haplosporids, rickettsias ference is greater than in Ensis arcuatus, which, as in type organisms, turbellarians, copepods, tremetode the previous case, may be due to its location. metacercarians and necrosis), in addition to the presence of the following parasites in both beds Immunological Study. As regards Ensis arcuatus a (Nematopsis sp. and coccidiae), however with a similar cell concentration was observed in the two higher prevalence in the Cies bed, and other para- beds (Cíes and Cambados) in 2005. In 2004, howev- sites (bacterial bags, ciliates, trematode sporocysts er, the concentration was higher in Cíes. The data and infiltrations) with low levels of prevalence regarding hemocyte viability showed similar results found to be similar in the two beds, although slight- in both beds. The data on cell concentration and via- ly higher in Cambados. The data obtained from the bility for Ensis siliqua were similar in Cangas and Ensis siliqua beds in Finisterre (affected) and Cangas Finisterre. (not affected) also exhibited the presence of certain pathologies that appeared only in the Finisterre Reproductive Cycle. The beds of Ensis arcuatus bed (bacterial bags, rickettsias type organisms, gre- (San Martiño and Means) and Ensis siliqua (Fisterra garines, turbellarians, metacercarians and trema- and Barra) wee sampled over the course of 2 com- tode cercarians and granulocytomas), the presence plete reproductive cycles, immediately following the of Nematopsis sp. and Coccidiae, in both beds, but Prestige oil spill (November, 2002). Ensis siliqua with a higher prevalence in Finisterre, in addition to exhibited similar results, with the reproductive cycle

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starting in winter. In both beds mature specimens Martiño, the period exhibiting maximum values last- began to be observed in late March (2003 and 2004). ed only from January to April, which would appear During the months of April and may, 100% of the to indicate that no spawnings occurred after this individuals were mature and had reached the date, which is an unusual occurrence in this species. spawning stage. In the Barra bed, a percentage of clams n the spawning stage were still detected until Larval Study. The larval cultures of Ensis arcuatus the beginning of July, whereas in Fisterra 100% of in Rodas (affected area) and Means (not affected) the animals had completed the cycle the previous did not show any significant differences in either month. On the basis of the gonad condition index growth or survival. However, the protein content in results and the histological preparations observed, it percentage and in µgr protein/µgr larva, in all stages was deduced that the two beds of Ensis arcuatus except larval stage D (1 day) was different, being sampled carried out consecutive spawnings from higher in Rodas. Three complete monitoring studies February-March, 2003 until June and remained in were carried out on Ensis siliqua in Fisterra. We still the sexual rest period from July to October. During need to take some more samples of the unaffected the following cycle, differences in the gonad condi- bed (Barra). Currently underway are the analyses of tion index were observed in both beds. This was due fatty acids, and so far no differences between the to the fact that in Meáns there was a succession of beds have been found. peaks from December to June, 2004, while in S.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Identification of genes as potential indicators of Abollo Rodríguez, E. resistance/susceptibility to diseases in the European flat oyster (Ostrea edulis) and in the RESEARCHERS: Pacific oyster (Crassostrea gigas). CIMA: Villalba García, A.; Cao Hermida, A.

START: August, 2005 INTERNS: FINISH: July, 2008 Comesaña Lestayo, P.

FINANCIAL ASSISTANCE: Dirección Xeral de Investigación e Desenvolvento. Consellería de Innovación e Industria. (Directorate General of Research and Development. Local Ministry for Innovation and Industry) Xunta de Galicia

SUMMARY Oyster farming worldwide is dominated by the of P. olseni, P. mediterraneus, P. marinus and P. culture of the Pacific oyster Crassostrea gigas chesapeaki. For the evaluation of the RFLPs, we (accounting for 97%of the world production), cloned and sequenced the PCR products. Research owing largely to its rapid growth rate, low mortali- done using the BLAST (Basic Local Alignment Search ty and the fact that it lends itself easily to aquacul- Tool) belonging to the servers of the National ture. This project aims to study the differences in Center of Biotechnology Information and the phylo- susceptibility to infections between the European genetic analyses (Minimum Evolution and Maximum flat oyster and the Pacific oyster. The objectives Parsimony analysis) confirmed the identification of included are the identification and characterization the species made by the RFLPs. This test makes it of genes induced in the European flat oyster and possible to detect different species co-infecting the Pacific oyster in the presence of infections. The first same host and it also is useful in the epizootiologi- question we address is the execution of a quick and cal studies currently underway, as well as in the dis- accurate diagnostic method to detect the main ease control programs. pathogens affecting the culture of the oyster to be able to differentiate between healthy and infected The differential diagnosis of bonamiasis was also oysters. This separation of healthy animals from carried out successfully, following the same strategy infected ones is crucial to the success of this project, as used to diagnose Perkinsus. By means of PCR we since this is the foundation on which the genetic amplified a fragment of the gene 8S of the DNAr libraries will be prepared for subsequent removal. using the primers BO and BOAS (Cochennec et al. The Polymerase Chain Reaction technique (PCR) is 2000). In cases where infection from Bonamia was used to make a differential diagnosis of the three detected, these primers produce an amplification of main etiological agents affecting the culture of the 300 bp which is easily visualized in 2% agarose gels oyster: Bonamia, Haplosporidium and Perkinsus, stained with ethidium bromide. The specific identifi- whose presence is compulsory to report as stipulat- cation of this etiological agent was carried out using ed by the international office of Epizootias. RFLPs. The endonuclease BglI produces a restriction pattern of two fragments of approximately 180 bp For the specific diagnosis of the different species and 120 bp for Bonamia ostreae and a single band of Perkinsus, we designed a PCR test followed by an of 300 bp for B. exitiosa. The results were evaluated analysis of the restriction fragment length polymor- by cloning and sequencing the PCR products. phism (PCR-RFLPs). The RFLPs analysis was carried Research using the BLAST confirmed the species out on an amplified fragment of the ITS region of identification carried out by RFLPs. RNAr, using restriction enzymes. These enzymes gave rise to species specific fragment patterns, For the specific diagnosis of haplosporidiosis, we which allowed us to successfully identify the species designed primers using a sequence of the gene 18S

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of DNAr of Haplosporidium armoricanun from O. Haplo-1517R, amplify a fragment of roughly 170 pb. edulis, kindly supplied by Dr. Burreson (Virginia The initial results obtained show that these primers Institute of Marine Science, USA). The primers were are specific to the diagnosis of haplosporidiosis and designed with the help of the Primer-3 program. do not amplify species that are phylogenetically These primers, which we call Haplo-1348F and related.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Bonamia ostreae life cycle investigations, optimised Hugh-Jones, D. production of resistant Ostrea edulis spat, and stud- ies of oyster (Ostrea edulis) immune mechanisms. CHIEF RESEARCHER AT THE CIMA: -Investigaciones sobre el ciclo de vida de Bonamia Villalba García, A. ostreae, producción optimizada de semilla de Ostrea edulis resistente y estudios de los mecanis- RESEARCHERS: mos de inmunización de la ostra (Ostrea edulis). CIMA: Cao Hermida, M.A.; Carballal Durán, M.J.; Casas Liste, S.M.; Fuentes González, J.M. START: April, 2003 FINISH: September, 2005 RESEARCH SUPPORT: Gregorio Chenlo, M.V.; Meléndez Ramos, M.I.; FINANCIAL ASSISTANCE: Penas Pampín, E. European Union, 5th Framework Program, Contract No. Q5CR-2002-72338 INTERNS: Comesaña Lestayo, P.; Rodríguez García, C.

SUMMARY This project has been coordinated by the Irish oys- the Ría de Arousa. As a reference for comparison, ter farming company ATLANTIC SHELLFISH LTD. and the experiment included oysters from two natural included the participation of a French company and beds in Galicia, one from the Ría de Ortigueira and a Galician enterprise (JOSÉ MARÍA DAPORTA LEIRO another from the Ría de Pontevedra (oysters adapt- E HIJOS S.L.), two oyster farming cooperatives, ed to the Galician environment) as well as oysters of (Dutch and Irish) and an association of British pro- the species Crassostrea gigas (resistant to bonamio- ducers, along with two research centers, “University sis). The oysters used at the time they were hung College Cork” (Ireland) and CIMA. The project focus- from the raft had a minimum age of two years, to es on fighting bonamiosis, a disease largely respon- ensure susceptibility to the disease. Annual mortali- sible for the decline in the flat oyster Ostrea edulis ty rate estimated for each strain of the European industry in Europe, constituting the main obstacle to flat oyster were as follows: Rossmore 75%, Tralee relaunching this industry. This project was born of a Bay 82%, Ortigueira 87%, Pontevedra 54%. Both previous project which demonstrated that a strain the prevalence and intensity of bonamiosis were of flat oyster called Rossmore is more tolerant of lower in the Rossmore oysters, which would confirm bonamiosis than oysters from other European popu- the fact that this strain is less susceptible to the dis- lations. The objectives of the project are: 1) to opti- ease. The highest prevalence values were found in mize spat production of the Rossmore strain, 2) con- oysters from Tralee Bay, where this disease was firm that this strain is more tolerant of bonamiosis never detected. and allows greater survival rates than native oysters during the growth process in several European A comparison of the immunological capacity was countries, 3) shed light on the biological cycle of the carried out on samples collected both before the dis- parasite Bonamia ostreae, with special emphasis on ease was detected and after the disease had mani- the stages developed outside of the flat oyster and fested itself. The parameters selected to compare 4) analyze the immunological mechanisms of the the immunological capacity among the different flat oyster and find differences between strains that oyster strains were the following: total number of are tolerant of and susceptible to bonamiosis, and hemocytes in the hemolymph, phaogcyte capacity which may serve as a basis for tolerance. The CIMA of the hemocytes, the production of toxic radicals of is participating in objectives 2 and 4. oxygen by oxidative metabolism (superoxide anion, hydrogen perioxide, nitric oxide), antibacterial activ- In 2003 Rossmore oysters and oysters from an area ity of the hemocytes and plasma, levels of phe- in Ireland where bonamiosis was not detected, noloxidase and phosphatase acid in the hemocytes Tralee Bay, -and therefore considered to be highly and plasma and protein stress synthesis after ther- susceptible to the disease-were hung from a raft in mal shock. No consistent differences were detected

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among the different oyster strains in the parameters granulocytes, It was also found that the infection tested that would shed light on their different sus- significantly inhibits the intra- and extra- cellular ceptibility to bonamiosis. However, significant dif- production of the superoxide anion by the hemo- ferences were found between O. edulis and C. gigas cytes of the oyster, which allows the parasite to sur- in some of the parameters, particularly the total vive in the hemocytes. number of hemocytes, the relative abundance of the different hemocytic types that could explain, at The histoptahological analyses would suggest that least partially, the differences in susceptibility O. edulis oysters are more susceptible to the pro- between these two species of oyster. gression of the B. ostreae infection when the gonad is in the female stage. The proportion of oysters Differences were also found between oyster with exclusively or predominantly female gonads infected with B. ostreae and those that were not was significantly higher in the strain that is more affected. They demonstrated that the infection susceptible to the disease (Tralee Bay). This would results in a substantial increase in the number of cir- suggest that selection against bonamisosis favors culating hemocytes as well as a rise in the relative male gametogenesis as opposed to female. abundance of small hialinocytes in detriment to the

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Optimization of the culture of the European flat Montes Pérez, J. oyster in Galicia by using quick diagnostic tech- niques to monitor the parasite Bonamia ostreae. RESEARCHERS: INTECMAR: Fernández Conchas, R.; Pérez Bello, J.L. START: March, 2004 FINISH: December, 2006 RESEARCH SUPPORT: CIMA: Meléndez Ramos, M.I.; Penas Pampín, E. FINANCIAL ASSISTANCE: INTECMAR: González Casal, G.; Alonso Fariña, B. Dir. Xeral de Inn. e Dsenvolvenmento Pesqueiro. Consellería de PAM. Xunta de Galicia

SUMMARY In Galicia, bivalve mollusk culture is of great ough control, classic histological techniques are also importance on both a social and economic level. The used as a supplementary test. In this way, we hope appearance of diseases has, on occasion, limited the to be able to provide oyster farmers with key infor- development of these cultures. A case in point is the mation to help them choose culture strategies and flat oyster (Ostrea edulis L.), which has suffered the most appropriate times to market the oysters. high mortalities associated with the presence of the parasite Bonamia ostreae (included in List II of In order to do this, controls are carried out to Community Directive 91/67/CEE) which causes the monitor the batches of oysters both on their arrival disease known as bonamiosis. to Galicia as well as during the subsequent culture, so that we will be able to inform the oyster farmers In recent years, owing to the lack of oyster spat regarding the levels of parasitization of the import- and the high mortalities caused by bonamiosis, most ed oysters and to devise culture strategies and the of the oyster farmers decided to import oysters of appropriate times for marketing. different sizes, although some nearly of commercial size, from different countries, to reduce the time Preliminary results indicate that the Polymerase needed for culture on the Galician coast. This prac- Chain Reaction technique (PCR) is a quick and sensi- tice entails a serious risk as it serves to strengthen tive method to detect Bonamia ostreae in the the parasite, while it does not aid in reducing the European flat oyster. It is also interesting to note disease. that most of the batches of oysters analyzed having an age of around 18 months and imported form This action aims to carry out routine tests using a France were infected with Bonamia at the time they Polymerase Chain Reaction technique (PCR) to facil- were introduced to Galicia, and that the mortalities itate the control and monitoring of the existing cul- detected in some cases were associated with the tures of the flat oyster in Galicia. To achieve a thor presence of this parasite.

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Development of a genetic selection program to Villalba García, A. produce a strain of flat oyster, Ostrea edulis L., resistant to bonamiosis. RESEARCHERS: CIMA: Carballal Durán, M.J.; Fuentes González, J.M. Criadero de Moluscos de Punta Quilme, Illa de START: January, 2004 Arousa: García Fernández, A. FINISH: December, 2006 RESEARCHER SUPPORT: Gregorio Chenlo, M.V.; Meléndez Ramos, M.I.; FINANCIAL ASSISTANCE: Penas Pampín, E.; Varela Manso, O.M. Dir. Xeral de Inn. e Desenv. Pesqueiro. Cons. de Pesca e As. Marítimos INTERNS: Xunta de Galicia. Comesaña Lestayo, P.; da Silva Scardua, P.M.

SUMMARY The disease caused by Bonamia ostreae curtails In February, 2005 conditioning began with the the development of the European flat oyster indus- oystes selected as parent stock. The purpose was to try in Galicia. In a previous study which evaluated obtain offspring consisting of (1) pure lines, (2) line- several families of European flat oyster reared in a ages resulting from the mixture of the pairs from hatchery using parent stock from different geo- selected families and (3) oysters from non-selected graphic regions, significant variability was found in parent stock collected from the natural beds and terms of susceptibility to the disease and survival used as a reference. The distribution of the oysters rate in the populations of European flat oyster and was as follows: 2 tanks containing 30 oysters each that prolonged exposure of a natural population to from a Greek family (GR5); 3 tanks each containing the disease might have resulted in a process of nat- oysters from a different family from the Ría de ural selection of tolerance to the disease. This back- Ortigueira (16 oysters from OR2, 30 oysters from ground information serves as a solid base on which OR3 and 24 oysters from OR4); 2 tanks each with to develop a genetic selection program that will oysters from a different family from Coroso (Ría de provide a quality strain of European flat oyster to be Arousa) (23 oysters from CO4 and 30 oysters from reared on an industrial level in Galicia. The aim of CO5); 4 tanks containing a mixture of oysters from this action is to obtain the first generation of two families from the Ría de Ortigueira, 6 from OR2 European flat oyster spat, using, as parent stock, and 6 from OR3 in each tank; 4 tanks each contain- oysters from families that have been identified as ing a mixture of oysters from two families from favorable in the previous study and to evaluate the Coroso, 8 from CO4 and 8 from CO5 in each tank. yield in raft culture. The original idea was to devel- Non-selected oysters were also used : 2 tanks each op the production of spat in an industrial hatchery. containing 30 oysters collected from a natural bed in These attempts, however, were unsuccessful – very the Ría de Ortigueira and 2 tanks each containing few broods were collected and the larvae that were 30 oysters from a bed in the Ría de Pontevedra. collected died only a few days after culture. In July, After standard conditioning, several larval broods 2004 the surviving parent stock were returned to were obtained in almost all of the tanks. The larvae the raft. This failed effort in the industrial hatchery, from different spawnings were cultivated separate- due to inappropriate management, led us to seek ly. The postlarvae were reared until they reached a out other facilities that would allow for the suitable size that was able to be retained in a 5 mm. sieve. A management of the parent stock and larval culture. high mortality rate was found during the grow-out We decided to remodel a small hatchery that was of the postlarvae. In September, 2005 all the batch- being set up by the CIMA, even though the condi- es of spat produced were moved to raft in the Ría de tions were precarious for the production of spat Arousa – amounting to a total of 4700 units distrib- from different lineages. uted in the following manner: 2 batches of spat

62 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

descending from “pure families”, OR2 and CO5; 3 measured, these oysters were returned to the raft. batches of spat descending from 3 different tanks In the last grow-out stage, histopathological analy- that contained oysters from a mixture of families ses were performed to determine the extent of OR2 and OR3; 3 batches of spat descending from pathological alternations. Owing to the lack of tanks containing oysters from a mixture of families appropriate facilities in the hatchery, the experi- CO4 and CO5; 2 batches of spat descending from ment suffered a one-year delay, and, as a conse- oysters from the bed in the Ría de Ortigueira and y quence, the number of batches of spat and individ- 2 batches of spat descending from oysters from the uals per batch was lower that what was originally bed in the Ría de Pontevedra. planned. Nevertheless, currently being cultured are 2 batches of oysters descending from pure families, The evolution of the spat on the raft was evaluat- 6 batches of oysters from the mixture of selected ed by carrying out quarterly estimates of the mortal- families with the use of marker microsatellite ity and growth of each batch of oysters. After being sequences.

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3.1.3. AREA OF COASTAL OCEANOGRAPHIC PROCESSES

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Depuration of diarrheic type toxins (DSP) in the Blanco Pérez, J.C. Mussel Mytilus galloprovincialis and amnesic type in the Great Scallop Pecten maximus. RESEARCHERS: CIMA: Pérez Acosta, C. START: August, 2004 INTECMAR: Moroño, A.; Salgado, C.; Arévalo, F.; FINISH: August, 2007 Correa, J.; Fernández, M.L.; Maroto, J.; Borras, G.

FINANCIAL ASSISTANCE: RESEARCH SUPPORT: Dirección Xeral de Investigación e Desenvolve- Mariño, C.; Muñíz, S.; Martín, H. mento. Consellería de Innovación, Industria e Comercio. (Directorate General of Research and INTERNS: Development. Local Ministry for Innovation, Mauríz, A. Industry and Commerce) PGIDIT04RMA501007PR

SUMMARY toxins in the mussel Mytilus galloprovincialis, but The accumulation in bivalves of toxins from phy- within which the specific objectives will adapt to the toplankton is probably the main problem in the cul- peculiarities of each organism/toxin combination: ture and harvesting of bivalves in Galicia as well as 1.- Environmental control and diet during the in other areas of Europe and the world. The toxins depuration process do not affect the different species being reared in 1.1.- Effect of temperature (ASP, DSP), dissolved aquaculture, in the same way. The toxin/species oxygen (ASP) combination having the greatest economic and 1.2.- Effect of anaerobic metabolism induced social repercussions in Galicia are the DSP type in the by emersion (DSP) mussel and the ASP type in the scallop. ASP toxins 1.3.- Effect of diet quantity and type (owing to require prolonged depuration times and DSP toxins its richness in non-degradable materials and appear frequently, and have recently been signaled contents of substances that can compete with out as potential carcinogens. Most strategies devel- the toxins or eliminate their receptors) (ASP, oped to eliminate, or at least mitigate, the conse- DSP) quences of these processes require a strict control 2.- Basic mechanisms for the elimination of toxins monitoring the presence of these toxins so that 2.1.- Balances of toxins (ASP, DSP) bivalves will accumulate lesser amounts of toxin or 2.2.- Ability to degrade organs or tissues (ASP, so that they will eliminate them rapidly. In order to DSP) do this, it is necessary to understand the mechanisms 3.- Distribution of toxins in the bivalves involved in these processes, their kinetics, and the 3.1.- Intracellular distribution (ASP, DSP) factors regulating them. Although significant 3.2.- Receptors of the toxins (ASP, DSP) progress has been made in these areas, the results 3.3.- Anatomical distribution (DSP) obtained to date have not been as efficient as we 4.- Biotransformation of the toxins would have hoped for, mainly because there is still a 4.1.- Biotransformations in each organ (ASP, great lack of knowledge about the basics of depura- DSP) tion and its regulation. 4.2.- Effect of the biotransformations in the kinetics of elimination (ASP, DSP) To solve this problem and open up new channels for the development of accelerated depuration sys- The results of the above will establish, at the very tems, we propose the following objectives, organ- least, the foundation on which the processes of ized into four groups that will be applied to the ASP depuration and accumulation will be based and will toxins in the scallop Pecten maximus and the DSP improve the possibilities of health control by con-

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tributing to the modification of the existing EU reg- facilities on land for 8 weeks. Samples were taken ulations. Methods of the direct application of depu- weekly (3 batches of 30 mussels) which were dissect- ration might also be found which would open new ed into two parts: the digestive gland and the possibilities for the development of strains that remaining soft tissues. The toxins were extracted undergo quick depuration or slow accumulation. from the samples and quantified. Some of the sam- ples selected were subjected to hydrolysis to deter- In keeping with the work plan of the project, the mine their content in derivatives of the principal following activities have been developed: toxins. The results showed that the quantity of DSP toxins in tissues other than the digestive gland was 1. DSP toxins in the mussel negligible. The analyses were completed to obtain 1.1 Experiment on the effect of emersion and diet more detailed information on depuration and toxin quantity: A) two experiments were designed and transformation. conducted, the first being a factorial experiment Taking advantage of the experiment described in taking into account 4 factors: Toxin concentration in section 1.2., we also analyzed the differences and the mussel, food supply prior to emersion, emersion anatomical distribution of DTX2 in the mussel. time and food supply after emersion. Mussels con- taining okadaic acid as the main DSP toxin were 2. ASP toxins in the great scallop 2.1 and 2.2 These used. At the end of the experiment, toxins were two tasks were combined into one, examining the extracted from the digestive gland of the mussel effect of the quantity of diet and the temperature in and the amount of toxins in the extracts was ana- one experiment, which includes the two above-men- lyzed by HPLC attached to a mass spectrophotome- tioned factors. Between-individual dispersion was ter. At the present time the quantification of the much higher that what had been previously estimat- derivatives of the toxins by alkaline hydrolysis is ed. Therefore, the results of this experiment are not pending the modification of this technique, since conclusive. the ones that have been published have not yielded satisfactory results. The second experiment focused In another experiment, we studied the effect of 4 on the effect of emersion time on the depuration of types of diet on two levels. Although the scallops DTX3 and its derivatives. The analytical phase is in were clearly affected by the different diets, the dif- the same state as the previous experiment. This ferences in the depuration of ASP toxins were small. experiment used mussels containing DTX2 as the main DSP toxin. A new experiment, dealing exclusively with tem- perature, using seven levels of this factor, is current- 1.2. An experiment of the effect of environmental ly underway. factors. A factorial experiment was conducted including temperature and diet quantity. The ana- 2.3 Balance of ASP toxins. The current techniques lytical phase is in the same state as the previous used to determine toxins in the water and the high experiments variability of toxin concentrations between individ- uals make it necessary to develop new techniques 1.3 Anatomic distribution of the toxins in the mus- or to modify the ones currently available. We are sel. Taking advantage of a toxic episode of DSP with working on this at the present time. okadaic acid , contaminated mussels were held in

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3.1.4. ÁREA DE RECURSOS MARINOS

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Introduction of immunodetection techniques to Fuentes González, J.M. monitor mussel larvae in Galician coastal waters. I. Adaptation of sampling protocols and larva identi- RESEARCHERS: fication. CIMA: Lorenzo Abalde, S.

START: August, 2005 RESEARCH SUPPORT: FINISH: August, 2008 Gregorio Chenlo, M.V.

FINANCIAL ASSISTANCE: Plan Galego de IDIT. (Galician Program fro Research and Technological development. Marine Research Sectoral Program).

SUMMARY The culture of the mussel, Mytilus galloprovin- larvae and postlarvae in plankton from the Ría de cialis, in Galicia was started in the mid-20th century Arousa, (1999-2001)”, “PGIDIT-CIMA-02/5, Optimi- with the installation of the first rafts in the Ría de zation of sampling protocols of mussel larvae in the Arousa. Since that time the mussel-farming sector Galician Rías, (2002-2003) and “PGIDT-CIMA-04/1 has continued to grow at a considerable rate and Evaluation of the larval abundance and recruitment has now become the most important production of of mussel spat Mytilus galloprovincialis on the aquaculture in Spain. This culture entails the grow- Galician coast”. Ever since these studies began, the out on rafts of juveniles obtained from both inter- main problem has been the development of a quick tidal populations of this species as well as postlarvae and reliable method for the identification and sepa- gathered by means of collector ropes hung from the ration of mussel larvae from the larvae of other rafts. Despite the fact that recent studies have bivalve mollusk species coexisting on the plankton. demonstrated the advantages of using collector This problem was tackled jointly by research groups ropes, both methods of collection are still being from the Area of Immunology at the University of used at the present time. However, collector ropes Vigo and the Area of Bioecology and Culture of the are fast becoming the method of choice in the mus- Mussel at the CIMA, under the Research Project sel sector and the use of this method has been pro- “PGIDT00MAR50101PR, Immunological and bio- moted by new legislation passed by the Galician chemical Identification of mussel larvae Mytilus gal- Fisheries Administration. Since 1999, for the purpose loprovincialis, (2000-2003)”. The results of this study of advising both the mussel sector on their strategy have provided us with monoclonal antibodies form for the installation of collector ropes as well as the mice called M36.5 and M22.8 (provisional number, Galician Fisheries Administration as to their regula- pending registration with the OEPM P200300629), tory policies, (Decree 406/1996), The Instituto which specifically recognized mussel larvae. Now Tecnológico para o Control do Medio Mariño de that these antibodies have been obtained, they Galicia (Technological Institute for the Monitoring must be introduced into the monitoring of the mus- of the Marine Environment) (INTECMAR; previously sel larvae, which continues to be conducted weekly CCMM) and the Center for Marine Research (CIMA), in the coastal waters of Galicia. In order to do this, have been issuing weekly reports on the abundance we have proposed this research project in which the of the different larval stages of the mussel in current sampling protocols in the water column, the Galician waters. These reports are based on data col- preparation of samples in the lab and the methods lected by the Bioecology and Culture of the Mussel used to sort and identify larvae, will be adapted and group of the CIMA from plankton samples taken in modified to be able to apply these new immunode- a consecutive series of research projects: “PGIDT- tection tools. From the start of this project in the CIMA-99/6, Spatial-temporal distribution of mussel month of August until the end of 2005, several pre-

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liminary experiments have been performed to The results of the immunodetection tests also determine the most suitable method to preserve the demonstrated that samples with the primary anti- samples and the most appropriate incubation time bodies M36.5 y M22.8 that were incubated for 5 of the larva samples with the primary antibody. The minutes did not differ significantly from the results results of these experiments have shown that the obtained in immunodetection tests using longer use of fixing agents to preserve the samples (both incubation times (120 minutes). This made it possi- formaldehyde and alcohol) have a negative effect ble to reduce substantially the time it took to con- on the immunodetection test, as they significantly duct entire immunodetection test. increase the number of false negatives recorded.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: A study of the population of the great scallop Sánchez Mata, A. (Pecten maximus) in the Galician Rías.. RESEARCHERS: START: August, 2005 CIMA: Molares Vila, J.; Sánchez Mata, A.G. FINISH: July, 2008 Empresa Pereira SA: Sánchez Mata; Chapela Portela, A. FINANCIAL ASSISTANCE: Dirección Xeral de Investigación e Desenvolve- RESEARCH SUPPORT: mento. Programa de Recursos Marinos. Consellería Giráldez Rivero, R. de Innovación, Industria e Comercio. Xunta de Galicia. (Directorate General of Research and Development. Local Ministry for Innovation, Industry and Commerce) PGIDIT 05RMA 023E

SUMMARY The natural beds of the great scallop (Pecten max- and viscera. This makes it impossible to market this imus) in the Galician Rías are affected by biotoxins, species as a fresh product and requires that the ani- caused by the consecutive yearly episodes of red mals be eviscerated (removal of part of the viscera, tides. The repercussions of this make it difficult to where most of the biotoxins are accumulated). It market this species as a fresh product and some- may even lead to the total ban on harvesting. times the beds must be closed for long periods of Owing to the great market demand and the large times, in compliance with European regulations. In amount of biomass in the rías –since this species has 2005, all the Galician Rías, with the exception of the not been harvested due to the closure of its fishery- Ría de Arousa, were closed to extraction and com- for several years now, an alternative to the tradi- mercialization. tional fishery has been proposed, consisting of the relocation of these mollusks to areas that are affect- The aim of this project is to assess the natural pop- ed very little or hardly at all by red tides, so that they ulations of the great scallop in the Rías of Ares, may undergo depuration and extensive culture until Arousa and Vigo, to determine the spatial distribu- they reach commercial size. tion, abundance and population structure of this species. These types of cultures have been carried out with varying degrees of success with other species of The ultimate objective of this project is to study commercial bivalves. As regards the great scallop, the feasibility of collecting scallop spat from natural several authors have examined the ecology of this beds affected by biotoxins and relocating them in species. However, there have been no studies of this areas less affected by red tides, until they reach com- scale dealing both with the population structure mercial size. We also plan to gain insight into the and dynamics of this mollusk, and the development optimum environmental conditions necessary for of a fisheries management model. the development and growth of this pectinid. Objectives Background Evaluation of the natural scallop populations by The great scallop (Pecten maximus) is one of the means of an analysis of the of the distribution, most highly valued marine resources in the Galician abundance and biomass of the resource. fishery sector. The current problem that harvesters 1.- The location and delimitation of the natural of this shellfish are faced with is due to the phyto- beds of the production of pectinids in the Rías of plankton blooms that periodically affect the Vigo, Arousa and Ares. Galician Rías as red tides. The resulting biotoxins, 2.- Determination of the distribution of the scal- mainly ASP and PSP, are ingested by this pectinid lop for the sustainable harvesting of the resource. and remain in variable concentrations in the muscle

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3.- Analyses of the abundance and biomass of the sampled in each raking operation, for each bed. In populations in each bed being surveyed. order to do this, the positions were recorded with a 4.- A biometric analysis of the stock of each bed, GPS at the beginning and end of each operation, in to be recorded as the initial state of the population. addition to the total length of the rake, the time 5.- Recording and analysis of the environmental elapsed and vessel speed during the sample collec- variables related to the hydrography and sediment tion. in each natural bed, affecting the life history of this - All the scallop specimens obtained in each oper- species. ation were hauled on board. The abundance and biometry data of all specimens were recorded for Methodology each set. A number of individuals were labeled and - Locate and delimit the production beds of pec- set aside for subsequent lab analysis of biomass, tinids on the basis of data from the production and biopolymeric carbon in the muscle and gonad and harvesting areas of the scallop where the fisher- toxin level. men’s association harvest this resource in all three - At the same time, sediment samples were collect- rias. ed from each bed to perform a structural granulo- - Conduct an intensive sampling survey in all three metric analysis of the bottom type and total organ- rías using the type of vessel and gear employed in ic matter content. Also recorded were data on oxy- scallop fishery (standard scallop rake) in order to gen, salinity and bottom water temperature. determine the state of the stock in each bed. The Samples of water and the sediment were taken for technique commonly used by the shellfish harvesters chlorophyll analysis as an indirect measurement of is adapted to be able to quantify the surface area the main potential food source.

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Geographic Information System oriented towards Molares Vila, J. the management of specific resources (SIGREMAR). RESEARCH SUPPORT: START: January, 2005 Rodríguez Ruibal, M.L. FINISH: December, 2005 INTERNS: FINANCIAL ASSISTANCE: Crego Mata, A. Dirección Xeral de Inn. e Des. Pesqueiro. Consellería de Pesca e Asuntos Marítmos. Xunta de Galicia (Directorate General of Research and Development. Local Ministry for Fisheries and Maritime Affairs).

SUMMARY The Geographic Information System oriented contrast, the free shellfishery zones are usually sub- towards the management of specific resources jected to permanent overfishing. The reason for this (SIGREMAR) began as an action developed in the situation is usually explained by the tragedy of com- CIMA, with the collaboration of the Shellfishery mon ownership. When captures are reduced, effort Service and biologists from the CPAM zone, and, is increased, causing a greater decline in catches, particularly, the technical assistance of the fisher- until the resource is completely exhausted. This men’s and shellfish harvesters’ associations of occurs when several individuals share a limited Galicia. In 2005 INTECMAR assumed this line of resource. investigation, carrying out the maintenance and exploitation of this information system, with the One way to avoid this situation is to grant shellfish technical management provided by CIMA. harvesters certain rights for restricted use (shellfish- ery permits) that may be given on an individual basis Shellfishery is carried out in beds having three dif- or assigned to collective organizations (fishermen’s ferent types of ownership: the intertidal beds, pri- associations). This measure would not be successful vately owned (shellfishery concessions) or fisher- if it were not accompanied by a planning system in men’s associations (shellfishery permits) and the sub- each entity. The planning of the harvesting is carried tidal beds which are the property of the Local out under the PEM programs Therefore, the disap- Ministry for Fisheries and Maritime Affairs (CPAM) pearance or poor quality of these programs would (free shellfishery zones). This difference is tremen- inevitably lead to situations of overfishing in the dously important in the way the territory is man- shellfish beds. aged, owing mainly to the degree of responsibility assumed by the shellfish harvesters. In concessions, Shellfishery management is different in every the holders assume total control of the manage- country, and may even vary between the different ment of the park. In the permit regime, a co-man- autonomous regions of Spain. The general trend is agement system is used by means of the shellfish to evolve towards co-management systems similar harvesting plans (PEM) under which the entities are to those carried out in Galicia under the PEM pro- responsible for exploitation and security, while the grams. These plans, however, have room for Fisheries Administration oversees the whole process, improvement in some important aspects such as to avoid overfishing. Lastly, in the free shellfish har- design, monitoring and evaluation to be able to vesting beds, management responsibility goes to increase the profitability of the harvest and ensure the Administration, which organizes the program by sustainability. imposing general regulations under the Plan Xeral de Marisqueo (General Shellfishery Plan). Since 1992, the year in which shellfishery manage- ment through the PEM programs started in Galicia, The production of parks in a concession is highly the management of these plans by the Fisheries variable, depending on the skill of the owner. In Administration has improved somewhat. However general, however, this system has a high yield. In the inappropriate use of the information systems is

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an obstacle that is difficult to overcome, yet essen- Currently all the fishermen’s and shellfish harvesters’ tial to improving the quality of the entire process. associations that have technical assistance, are Therefore, in our opinion, it is necessary to redesign equipped with this tool, accounting for over half of this process to ensure the correct use of the avail- the Galician fishermen’s associations. able resources, to reduce processing time, improve program assessment and facilitate quality control. The use of this computer-based tool will help stan- dardize the model of the plan, but it cannot guaran- First of all, we must ensure the harmonization of tee the harmonization of the plans. To be able to the PEM model. It is unacceptable for some plans to achieve a substantial improvement in the plans, it is limit themselves to listing objectives, while others necessary to prepare a book detailing the proce- fill more than eighty pages with irrelevant or useless dures that will help technicians involved in the man- information in practical terms. This team has devel- agement of shellfish resources to become familiar oped a computer application called SIGREMAR that with and sue the best techniques currently available allows any type of PEM program to be drawn up, for stock assessment and the study of population using a standard structure, with certain required dynamics. At the present time we are working on fields, and others that are optional, depending on this point. the resource or availability of the information.

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Action plan for the recovery of the Bohído zone. Sánchez Mata, A.

START: February, 2005 RESEARCHERS: FINISH: December, 2006 CIMA: Carballal Durán, M.J.; Cao Hermida, M.A.; Molares Vila, J.; Villalba García, A. FINANCIAL ASSISTANCE: INTECMAR: Fernández Conchas, R.D. Dirección Xeral de Recursos Mariños. CETMAR: Parada Encisa, J.M. Consellería de Pesca e Asuntos Marítimos. Xunta de Galicia (Directorate General of Research RESEARCH SUPPORT: and Development. Local Ministry for Fisheries and Carreira Vázquez, P.; Crego Mata, A.; Maritime Affairs Darriba Santiago, C.; Martínez Verde, G.; Rey García, C.; Rodal Mallo, M.

INTERNS: Varela Sóñora, M.T.

SUMMARY The natural bed “O Bohído”, located between the the continuity and effort needed to consolidate this east coast of Arousa Island and the coast of Vilanova endeavor. y Cambados, has a surface area of 4.26 km2 . Similar to the bed in the “Lombos do Ulla” zone, Years ago the “O Bohído” zone was one of the this natural bed is classified administratively as “free most important shellfish beds in Galicia for both the shellfishery zone” which means that vessels belong- European flat oyster (Ostrea edulis) and the pullet ing to all the fishermen’s associations can operate in carpet shell clam (Venerupis pullastra). However, the area. This makes it difficult to implement actions mortalities caused by Bonamia ostreae in the oyster aimed at improving the exploitation and monitoring and undetermined causes in the clam, frequently of the harvesting activity, both during times when reported by the fishermen’s associations, and the harvesting is banned as well as when it is permitted. general overfishing that this bed was subjected to have led to a progressive decline in productivity. The good results of the Action Plan for the Recovery of the Lombos do Ulla zone, which are evi- In accordance with recent technical reports drawn dent from the progressive increase in the productiv- up by biologists from the Delegación Comarcal de ity and profitability of this bed, along with the la Consellería de Pesca e Asuntos Marítimos objective of the Local Ministry for Fisheries and (Regional Delegation of the Local Ministry for Maritime Affairs to obtain a general reference pro- Fisheries and Maritime Affairs) in Carril, at the pres- tocol that will optimize the management of shellfish ent time among the mollusk species of commercial beds in Galicia, would recommend the adaptation interest found in this bed are, first in order of impor- and gradual application of the methodology used tance, the pullet carpet shell clam, and as secondary there to other free shellfishery zone having differ- species, (Dosinia sp.), (Venus verrucosa) and ent environmental conditions and target species. (Venerupis aurea). Also present are the short necked clam (Ruditapes philippinarum), the scallop The Bohído bed was selected for different rea- (Chlamys varia), the razor clam (Ensis ensis), the sons: its great economic potential, the fact that it clam (Venerupis romboides), the cockle presents characteristics different from those in (Cerastoderma edule) and the razor clam (Solen Lombos do Ulla, and, owing to its location, it would marginatus). be favorable to the development of a protocol for the supervision and monitoring of harvesting activity. In recent years, with the collaboration of the fish- ermen’s associations in the area, several attempts The general objective of this Plan is the design have been made to recuperate this bed, but without and progressive application of a management pro-

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tocol in the Bohído zone that will optimize its pro- One aspect that is particularly relevant to the duction and profitability and, along with the Plan management of shellfish beds, in both the free underway in Lombos do Ulla, will serve as a refer- shellfishery zone regime as well as shell fishery with ence for the management of free shellfishery zones authorization, are the high mortalities in clams fre- in Galicia. quently reported by the fishermen’s associations in different areas of Galicia. In most cases there is no Responsible management of shellfishery resour- quantitative estimate available of the magnitude of ces must be based on a good, solid knowledge of the above-mentioned mortality. Nor is there any the environmental, biological and demographic information on the patterns of temporal (seasonal) aspects of the zones and species targeted for variation of the mortality of clams in Galicia due to extraction. However, in addition to this knowl- natural causes. In these circumstances, it is difficult edge, it is necessary to have a management team to determine whether the mortality reported is an able to analyze the evolution of the social and extraordinary phenomenon or not and to know the commercial aspects, and bear them in mind, to be extent to which it may be related to the manage- able to design the most suitable harvesting strate- ment being carried out in this production zone. gies at all times for the sustainable development of the economic profitability of the bed. It is also A plan of this type where environmental, bio- important for the sector to become involved and logical and management aspects are monitored to pursue the continuity of the Plan –and its initial provides an exceptional framework within which financial support- after it has ended. to work on identifying the mortality in clams, its causes, and to compare them with other produc- tion zones.

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Evaluation of larvae and recruitment abundance of Fuentes González, J.M. mussel spat Mytilus galloprovincialis, on the Gali- cian coast. RESEARCHER CIMA: Molares Vila, J. START: January, 2004 FINISH: January, 2007 RESEARCH SUPPORT: Andrade García, M.; Fernández Abuín, I.; FINANCIAL ASSISTANCE: Gregorio Chenlo, M.V.; Rodríguez Díaz, R. Dirección Xeral de Recursos Mariños. Consellería de Pesca e Asuntos Marítimos. (Directorate General of INTERNS: Marine Resources. Local Ministry for Fisheries and Lorenzo Abalde, S. Maritime Affairs.) Xunta de Galicia

SUMMARY Mussel culture is based on the grow-out –under tion available to be able to make decisions”. The raft culture conditions- of wild spat procured by aim of this research action is to continue to meet means of the extraction of juveniles attached to these needs by collecting analyzing the temporal open rocky coasts of the intertidal zones and by the data series on the abundance of the different larval collection of larvae in the inner ría zones using col- stages of the mussel in the water column and setting lector ropes hung from the rafts. The second out to obtain the data series related to the recruit- method of spat collection for mussel culture has ment of the spat of this mollusk in rocky intertidal become the method of choice among the mussel zones and on collector ropes. In the short term, this farming sector and has been strongly supported information will allow us to assess both the mussel through new legislation by the Galician Fisheries farming sector as regards the strategy used to Administration. Starting in 1999 and for the purpose obtain spat as well as the Galician Fisheries of advising both the mussel sector on the strategy Administration in terms of their making the decision to use for spat collection as well as the Fisheries to open and close the harvesting periods and the Administration itself in terms of its regulatory poli- natural collection of mussel spat (Decree 406/1996). cy, the CIMA has been issuing weekly reports on the In the medium and long term, the data obtained abundance of the different larval stages of the mus- will allow us to increase our knowledge on the pop- sel in the waters of the different sub-zones where ulation dynamics of the mussel during its early the mussel raft parks are located in the Galician Rías. stages of life and, therefore, increase our ability to These reports are based on samplings carried out in predict the biological processes involved (spawning the context of the research project and research seasons and number of broods, larval dynamics, set- action (“PGIDT-CIMA 99/6 Spatial-temporal distribu- tlement and recruitment). tion of mussel larvae and post-larvae in the plank- To meet these objectives, in 2005 weekly samples ton of the Ría de Arousa” and “PGIDT-CIMA 02/5 were taken of plankton at 13 points in the Galician Optimization of the sampling protocols of mussel rías (stations V1, V2, V3 in the Ría de Vigo; P2, P3 in larvae in the Galician Rías”, respectively). As these the Ría de Pontevedra; A3, A4, A9 in the Ría de reports have proved to be useful to both the mussel Arousa; M2 and M5 in the Ría de Muros; L1, L2 and culture sector and the Administration itself, the L4 in the Ría de Ares-Betanzos, of the monitoring Department of Marine Resources, in its report of 29 network at INTECMAR), on the basis of which the September de 2003, considers that ... “once the util- abundance (larvae/cubic meter) was determined of ity of the actions have been validated and the mon- the different larval developmental stages of the itoring techniques have been perfected, it is essen- mussel (larvae in D and larvae with an “eye”). Also tial that said actions be transformed into a “moni- from July to December, 2005 evaluations were car- toring” process and that on a routine basis technical ried out on the recruitment of mussel spat (mejilla) information must be available at both the opening in the rocky intertidal zones of 6 locations of the and closure of the campaigns, as well as in all cases Galician coast (A Guarda, Cabo Udra, San Vicente, where it is necessary to have such technical informa Monte Louro, Merexo and O Portiño). For this, the

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number of individuals attached (recruits), their cov- spat on the collector ropes by determining the num- erage and biomass were determined in monitoring ber of individuals recruited per meter of rope, their surface areas of 1 square meter (3 surface monitor- mean size and biomass, in two series of collector ing areas for each location), which had under gone ropes set out in 12 different culture parks in the rías previous preparation (June,2005) consisting of of Ares-Betanzos (2), Muros (3), Arousa (4), scraping and eliminating the existing biomass. Pontevedra (2) and Aldán (1). Finally, we also assessed the recruitment of mussel

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RESEARCH ACTION STAFF

TITLE: CHIEF RESEARCHER: Mapping and evaluation of areas and species of Molares Vila, J. interest in the action plan for the recovery of the “Lombos do Ulla” zone. RESEARCHERS: CIMA: Santos Piñeiro, I.; Sánchez Mata, A. START: August, 2002 CETMAR: Parada Encisa, J.M. FINISH: December, 2006 RESEARCH SUPPORT: FINANCIAL ASSISTANCE: Carreira Vázquez, P.; Crego Mata, A.; Dirección Xeral de Recusos Mariños. Consellería de Darriba Santiago, C.; Martínez Verde, G.; Pesca e Asuntos Marítimos. Rodal Mallo, M.. (Directorate General of Marine Resources. Local Ministry for Fisheries and Maritime Affairs.) INTERNS: Xunta de Galicia Varela Sóñora, M.T.

SUMMARY The action called “Mapping and evaluation of 8.94 and 10.33 ind/m? from 2002 a 2005. In the short areas and species of interest” included in the Action necked clam, the evolution in density was similar to Plan for the recovery of the “Lombos do Ulla” zone that of the carpet shell clam; 1.06; 0.42, 4.04 e 12.12 was initially programmed to end in December, 2004. ind/m? during the September-October samplings However, in view of the results obtained, the CPAM from 2002 to 2005. decided to extend this project for at least two more years. The cockle is the main species harvested in the Lombos do Ulla bed. The evolution of the harvesting In 2005 direct evaluations of the stock of this bed of this species from the start of this actions shows were carried out twice – the first in the month of the good results obtained. In subsequent shellfish- May after the 2004-2005 harvesting campaign had ery campaigns carried out from 2002 until 2005, this ended and the second, in the month of September, species was harvested in quantities of 169.8; 551.6 prior to the design of the 2005-2006 harvesting cam- and 765.3 Tm. Improvements in the management, paign. In June a third sampling was carried out to monitoring and supervision of the resource are con- quantify the spring recruitment of the target sidered to be the main reasons for its increased pro- species. The daily recording of temperatures and duction. salinities continued at 8 fixed points in the estuary under three different tidal conditions. Samples were A study of the spatial distribution of this resource also collected by the pathology and contamination over time showed the sporadic movement of this teams of the CIMA and INTECMAR, collaborating on species towards the outer estuary zone. Several lab this project. experiments have demonstrated the relationship between different environmental parameters such The results of the stock estimations over the as salinity and currents and the fact that the cockles course of these consecutive sampling campaigns come to the surface and are dragged away. These during the months of September-October from 2002 movements would explain the sudden disappear- until 2005 exhibited a stabilization of the density of ance of the resource, formerly interpreted as being the cockle in the more recent campaigns: 91.9, due to massive mortalities. By mapping the destina- 278.9, 303.9 and 209.8 ind/m? in 2002, 2003, 2004 tion areas of the cockles in movement, it is possible and 2005, respectively. The density of the pullet car- to improve the patterns for harvesting manage- pet shell clam increased during the first three years ment. At the same time, it was demonstrated that (3.6; 5.9 and 10.65 ind/m? in 2002, 2003 and 2004 monitoring the environmental variables in the zone respectively) and underwent a slight decrease in allowed us to predict these movements. September-October, 2005, with a mean value of 7.34 ind/m2. The carpet shell clam increased continuously The carpet shell, pullet carpet shell and short in the last four years with an evolution of 0.78; 0.33; necked clams are considered to be accompanying

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species in this bed. Owing to the reduced spatial dis- Over the course of 2005 the size of the specimens tribution and lower densities of these species, the caught during the 2004-2005 was monitored week- estimations of their abundance are less consistent ly and those from the 2005-2006, every two weeks. than that of the cockle. To obtain a more accurate The information used in harvest management, as estimations, we designed a stratified sampling strat- related to the change in zones and catch quotas egy based on sedimentology and bathymetry included the following: size evolution obtained on parameters. The results of this new strategy are cur- the basis of this monitoring activity, catch fluctua- rently being analyzed, although more sampling is tion per unit of effort established by means of the necessary. supervision system and the evolution of the first-sale prices as well as mapping the distribution of the Catches of the carpet shell clam increased over the resources. course of the three harvesting campaigns completed as of the beginning of this action. Starting at a catch The monitoring system for harvesting and the of 114 Kg during the 2002-2003 campaign, the fol- supervision of the catches have proved to be an effi- lowing campaigns resulted in a harvest of 15.1 and cient tool for the recovery of the free shellfishery 10.1 Tm. The short necked clam exhibited a continu- zone of Lombos do Ulla. In Both the systems for the ous rise in catches: 301 Kg in the 2002-2003 cam- supervision and monitoring of the bed allowed us to paign; 5.8 Tm in 2003-2004 and 10.6 Tm in 2004- gain insight into its specific production, which was 2005. The pullet carpet shell clam, in contrast, exhib- unknown up to now or recorded on a joint basis ited declining catches: 11, 8.8 and 3.1 Tm in the with the other shellfish beds. three consecutive campaigns. To date, there has been no plausible explanation for this diminishing production. In mid-2005 an experiment began with the objective of determining the natural mortality and growth rates of this species in the zone.

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SCIENTIFIC ACTIVITY

B.- PROJECTS IN WHICH THE CHIEF RESEARCHER BELONGS TO ANOTHER INSTITUTION

PROJECT STAFF

TITLE: CHIEF RESEARCHER: Development of a quick diagnostic method to López Romalde, J. (USC) detect brown ring disease and to discover new bac- terial pathogens in the clam. RESEARCHERS: CIMA: López Gómez, C. START: December, 2003 USC: Magariños Ferro, B. FINISH: December, 2006 RESEARCH SUPPORT: FINANCIAL ASSISTANCE: Meléndez Ramos, I.; Penas Pampin, E. Ministerio de Ciencia y Tecnología. Plan Nacional de I+D Nacional. (Ministry of Science and Technology. INTERNS: National R&D Plan.) Beaz Hidalgo, R. Código de Identificación (ID code): AGL2003-09307-C02-01

SUMMARY Over the course of this project objective 1 was detected and to study the histological alterations achieved (Design and evaluation of a quick, genetic found in these specimens. The characterization of diagnostic method, based on polymerase chain reac- the fermentative strains is nearly finished, with a tion (PCR), to detect the presence of Vibrio tapetis, high prevalence of different species of the genus the main bacterial pathogen in the clam). In keeping Vibrio being found. with this, we designed, optimized and validated a PCR protocol for the detection of V. tapetis, based The team from the University of Málaga has on the amplification of the gene 16S rRNA by means begun a study on the possible pathogenicity of the of specific primers for this bacterial species. The different fermentative bacterial groups found. method showed a specificity of 100% and the detection limit obtained was 10.5 pg/tube of PCR, During the analysis of the microbiot associated which is equivalent to 3-4 bacterial cells. with the clams sampled, we observed a low inci- Additionally, the PCR protocol was evaluated for the dence of V. tapetis, which, was, however, detected direct detection of the pathogen in clam tissue with the PCR protocol developed in objective 1 of infected experimentally. In this case a high sensitivi- this project. In fact, in all the samplings carried out ty was obtained which made it possible to detect the to date, it was only possible to isolate two strains pathogen in infected clams with doses of under of V. tapetis. This situation prompted us to ques- LD50 of the pathogen only a few days after the tion the effectiveness of the bacteriological culture infection. These results have been corroborated by media commonly used in sampling to isolate V. those obtained using natural samples in which the tapetis. Despite the fact that this study was not pathogen was detected with this procedure and not originally included in the project, we considered it with traditional bacteriological techniques. to be important since if the media used did not Therefore, a powerful method for the detection of work with this bacterial species, then the incidence brown ring disease in clams has been devised. of the pathogen in clam populations could be underestimated. It was observed that the TCBS The protocol developed led to a patent applica- (selective-differential medium for vibrios) and, in tion (P200302512). general, all the culture media containing biliary salts, largely inhibit the growth of V. tapetis and We also collected monthly samplings of clams therefore, if used, would hinder, to a great extent, from four Galician fishermen’s associations the detection of this microorganism. The medium (Redondela, Camariñas, Vilaboa and Carril), which that produced the best results for the detection allowed us to characterize the main bacterial groups and isolation of the pathogen was marine agar supplemented with manitol.

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In addition, a histopathological study on the pop- 2) Rickettsiano type intracellular colonies in the ulations of Ruditapes decussatus and Ruditapes branchial and digestive epithelium. philippinarum analyzed was carried out. After 3) Haplosporidium type plasmodia in the digestive recording the morphometric indices (length and live epithelia. weight), the clams were opened and subjected to a 4) Perkinsus atlanticus found in different organs. macroscopic examination to investigate the pres- 5) An unidentified gregarine in the intestinal ence of any anomaly, such as white pustules in the epithelium and oocysts of the gregarine visceral mass (sometimes associated with Perkinsus Nematopsis sp. in the gills. sp.), macroparasites, lesions in the gills, chambers in 6) Free ciliates on the surface and in the branchial the shell, brown ring, etc. Next we made a sagital water canals. cut of a fragment containing the largest number of 7) Turbellarians of the Paravortex type in the organs possible to be processed in a histological digestive lumen and of the Urastoma type in the analysis using conventional methods such as hema- gills and paleal cavity. toxyline of Harris-eosine stain. 8) Possible trematode metacercaria in the foot. 9) Hemocyte infiltrations and granulocytomas The parasites and/or alterations observed to date always associated with the presence of P. atlanticus. are as follows:

1) Bacterial bags in the branchial water tubes (around 100 µm) inside a fibrous covering coming from the conjunctive tissue of the base of the branchial epithelium.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Technology development for a reliable supply of Pauline Kamermans (RIVO-Netherlands; high quality seed in blue mussel farming. Project Coordinator)

START: November, 2005 CHIEF RESEARCHER AT THE CIMA FINISH: October, 2007 Fuentes González, J.M.

FINANCIAL ASSISTANCE: RESEARCHERS: 6TH European Union Framework Program. CIMA: Villalba García, A. Contract no. COOP-CT-2005-017729 (BLUE SEED) RESEARCH SUPPORT: Gregorio Chenlo, M.V.

INTERNS: Domínguez Pérez, L.

SUMMARY In addition to the serious damage caused by recur- Kingdom, France and Spain), aims to confront these rent episodes of toxic microalgae, there are two two problems in an attempt to find a solution. The main problems affecting the culture of the different project will hone techniques to rear mussels in the mussel species on the European coasts: (1) problems hatchery to guarantee producers a reliable supply related to the supply of reliable spat for culture and of top-quality spat. (2) reduced quality of the product during spawning periods. The first problem, - low and unpredictable The research project is structured into five work recruitment of mussel spat from both natural popu- blocks or packs. The first one establishes the pat- lations as well as collector devices- mainly affects terns for the internal organization for its implemen- European countries located more to the north tation. The second block deals with refining the (United Kingdom, the Netherlands, etc.). The sec- techniques for handling specimens to be used as ond, i.e., the loss of biomass, and therefore reduced parent stock as well as larva culture. The third block quality of the product, as a consequence of extend- will analyze different methods to procure triploid ed periods of spawning, mainly affects the countries individuals. In the fourth block, we will work with to the south of the continent (France, Spain, etc.). methods for spat collection and grow-out in both land-based facilities (seedbeds) and sea-based struc- This cooperative European project, with the par- tures. And the fifth area will focus on an analysis of ticipation of companies and research centers from the financial cost of the different production sys- four European countries (The Netherlands, United tems under analysis.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Emergency response to coastal oil, chemical and Simon Wilkins (Devon County Council, U.K.) inert pollution from shipping. RESEARCHERS: START: 2004 CIMA: Blanco Pérez, J.C. FINISH: 2007 CETMAR: Chapela, R.; Fernández, M.; Pungin, B. FINANCIAL ASSISTANCE: INTECMAR: Ayensa, M.G.; Molares Vila, J.; Montero, P. European Union CPAM: Novas, A.; Novoa, F. (INTERREG IIIB: Atlantic Zone program) Meteogalicia: Pérez, V. and other researchers from Portugal, France and the U.K.

RESEARCH SUPPORT: CIMA: Giráldez Rivero, R. INTECMAR: Gómez, A.; Rodríguez Ruibal, M.L.

SUMMARY In view of the challenges facing regional and local The coastal response to oil, chemical and inert pol- governments, as well as the organizations that pro- lution from accidental spills is probably one of the vide assistance, when confronted with pollution greatest challenges in terms of resources and admin- from shipping, the Association, EROCIPS has devel- istration facing regional and local governments, as oped a project with European Partners in France, well as the organizations that provide assistance Spain, Portugal, Ireland and the U.K. to provide a (governmental agencies, academic institutions, Non- suitable response to combat oil, chemical and inert governmental groups, representative organizations, pollution. etc.), participating in the spatial planning of the sus- tainable use of coastal assets. The Atlantic Zone of the European Union has been the site of a number of spills –all-too widely- Pollutants represent a direct threat to coastal known- over the last thirty years, which include: the assets such as protected areas, beaches, shellfish Amoco Cádiz, Betelgeuse, Aegean Sea, Sea Empress, hatcheries, ports, marinas and nature. The indirect Erika and Prestige. Each one of these spills has aftermath of a pollutant event may also be harmful, demonstrated the pressure exerted on the resources leading to public mistrust, affecting the tourist trade and on the administrative structures of the region- and income from fishing, which has a negative al and governments, in their attempts to limit the impact on the economy of the contaminated coastal impact caused by the pollution of the coastal assets area. of a zone. Despite the damage caused, each one of these spills has also provided participants with expe- To take on this challenge, the responding organi- rience in terms of the measures to be taken when zations have developed a structured response, gen- faced with an event of this type. A number of erally as part of joint local and national contingency regional and local governments of the Atlantic plans. These are based on the following: Zone, along with the organizations that provide - Protection of coastal assets (estuary, ports, aqua- assistance, now have the necessary foundations to culture, etc.), usually by means of floating barriers/ be able to develop this experience acquired in dams; response to these events into a tool. - Cleaning beaches; - A response favoring the protection of nature; The aim of the project is to formulate a transfer- - Transport of waste, able methodology able to communicate the perti- - Provisional storage of waste . nent information to the participants who are in a position to respond to and make decisions in opera- The underlying theme of all of these activities is tions to combat coastal pollution from shipping. the need to guarantee the safety of the participants

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in the coastal action and citizens alike. In some cir- pollution events and their impact on the coast. ERO- cumstances, for example, the risk of toxicity or CIPS has laid down four objectives to meet these explosion may warrant the massive evacuation of requirements and contribute to the final goal: local communities. - Develop common tools and techniques to identi- fy the position of reference and to classify risk or The response may be less effective if the organiza- sensitive points on the coasts; tions are not able to avail themselves of the appro- - Develop common tools and techniques to identi- priate techniques, sufficient resources or if the fy all the necessary information for the effective administrative structures do not have the necessary planning of a coastal response to combat coastal capacity. The key to guaranteeing the efficiency of a pollution events on the coast; response is to provide updated information to the - Formulate common advice, standards and proto- action groups through a concentrated administra- cols to handle coastal pollution events, that are tive structure. The EROCIPS Project provides this transferable to all the coastal zones of Europe, information. - And link the framework of action to combat coastal pollution events to other spatial planning The Project, Emergency Response to Coastal Oil, structures on a local and regional level to guarantee Chemical and Inert Pollution from Shipping (ERO- a coordinated and effective response. CIPS), consists of a series of work packs designed to provide coastal action groups with the necessary The EROCIPS project does not assume statutory information to guarantee a response centered on obligations related to the formulation of contin- combating pollution. The aim is to offer a procedure gency plans as established by the member nations. that can be applied to other coastal zones in Europe. The purpose of the project is to provide an added value to the coastal response by supplying informa- The objective of the EROCIPS Project is the «devel- tion to regional and local governments to expedite opment of common transnational methodologies, the structuring of the response in the event of pol- tools and techniques to be used in response to lution from shipping. coastal pollution from shipping, transferable to the entire EU, in order to support the sustainability of Measure B-1 requires the development of efficient maritime transport systems ». and sustainable transport systems. EROCIPS takes a direct approach to this issue. The sustainable devel- In order to do this, regional and local govern- opment of international or coastal maritime traffic ments must have a common understanding of the in the Atlantic Zone requires efficient skills to deal many socio-economic and environmental factors with maritime pollution of any type or scope in that will contribute to an effective response. This order to protect the economic, environmental and includes the comprehension of a reference base social resources on land. EROCIPS will give region- along the coast and the tools and techniques than al and local governments, as well as the organiza- can be effectively applied according to the different tions that provide assistance, the ability to provide a situations and settings. Also involved is the effective quick, effective and real response on land, because integration of the response information from other it has the appropriate tools, protocols, methods, zones of spatial planning to achieve a coordinated procedures and information needed to plan and action response and an understanding of the coastal successfully execute this goal.

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Gradients of biological diversity in intertidal Mora Bermúdez, J. (USC) mobile substrates of the Antarctic: Connections between Antarctic and Sub-Antarctic communi- RESEARCHERS: ties (BINTART). CIMA: Sánchez Mata, A. UV: Lastra Valdor, M.; García Gallego,M. START: October, 2002 Universidad Valdivia, Chile: Jaramillo Lopetegui, E. FINISH: September, 2005

FINANCIAL ASSISTANCE: Ministry of Science and Technology Ntional R&D&I program REN2002-03109

SUMMARY A comparative analysis is being carried out on the erishment or enrichment) in terms of the degree of gradients of biological diversity of the macrofauna environmental stress (hydrodynamism and variation inhabiting intertidal mobile substrates of the in sediment composition) to which they are subject- Antarctic and Sub-Antarctic regions. This study ana- ed. The results obtained will be interpreted from a lyzes the biological diversity on three scales: local, macroecological and evolutionary standpoint, in an geographic and temporal during three sampling sur- attempt to analyze the impact of environmental vari- veys carried out in 2003 and 2004. ables in Antarctica on ecosystems with low diversity. 1) Being studied on a local scale are the patterns of biological diversity in the benthic communities Objectives: inhabiting the intertidal zones of Livingston Island, The general objective of this research project is to Deception Island and the Sub-Antarctic Magellan carry out a comparative study on the gradients of Region (Punta Arenas). The local gradients were biological diversity on a local, regional and temporal established in terms of tidal level, degree of expo- scale of the intertidal benthic communities inhabit- sure to waves, nature of the sediment and morpho- ing the mobile substrates of the Antarctic dynamic characteristics (tidal slope, hydrodynamism (Livingston Island and Deception island) and Sub- of the surveyed zone, etc). The sampling areas cov- Antarctic regions (Magellan Region: Punta Arenas). ered the widest possible range of environmental 1.- Study the gradients of biological diversity on a conditions, to encompass the broadest possible local and regional scale of the intertidal mobile sub- diversity of habitats. strates of the Antarctic and Sub-Antarctic regions. 2) Being studied on an intermediate geographic This main objective can be broken down into the scale, are the existing diversity gradients between following specific objectives: the islands of Livingston and Deception. • 1.1.- Analysis of the communities.- A compa- 3) On the scale of latitude, we are studying the rative study of the taxonomic composition, diversity gradient and the evolutionary patterns of abundance, biomass and diversity of the ben- speciation on both sides of the Antarctic conver- thic macrofauna of the intertidal mobile subs- gence, i.e., between the Antarctic region (Islands of trates of the Antarctic and Sub-Antarctic Livingston and Deception) and the Sub-Antarctic regions on a local scale (tidal zone and mor- region (Magellan Region of South America : beach- phodynamic and sedimentological gradients in es of Punta Arenas). each location) and a regional scale (between 4) The variability study on a temporal scale locations). involved a new sampling survey during the 2004 • 1.2.- Physical and morphodynamic analysis.- A Antarctic expedition. Patterns of ecological succes- comparative study of the physicochemical con- sion were studied in the Antarctic and Sub-Antarctic ditions (Temperature, salinity Chlorophyll a, regions, and the temporal variability of the commu- organic matter, sediment characteristics, etc.) nities was interpreted (substitution, species impov and morphodynamics (morphology, hydrody-

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namism) underlying the local and regional • 2.1.- Interannual variation: The Community. diversity gradients. An analysis of the interannual variation of •1.3.- Diversity Function.- A study of the role of taxonomic composition, abundance, biomass biological diversity, redundancy and functional and diversity of the macrofauna inhabiting the exclusivity of the species in the Antarctic and mobile substrates of the Antarctic and Sub- Sub-Antarctic zones, using a biogeographic Antarctic regions. and evolutionary focus. • 2.2..- Interannual Variation: The physical and morphodynamic environment.- An analysis of 2.- A study of the temporal variability of the com- the interannual variation of the physicochemi- munities inhabiting the mobile substrates of the cal and morphodynamic conditions of the areas Antarctic and Sub-Antarctic regions. This objective under study. involves carrying out a sampling survey just like the • 2.3.- Interannual Variation: diversity gra- one conducted to meet objective 1, which will be dients.- Interpretation of the temporal variabi- done during the second year of the project. lity of biological diversity from a biogeogra- Similarly, this main objective can be broken down phic and evolutionary standpoint. into the following specific objectives:

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Development of elements, tools, protocols for González-Garces Santiso, A. (IEO-Vigo) action and an information system for the design of contingency plans in the event of accidental marine RESEARCHERS: spills (CONTINMAR). CIMA: Molares Vila, J. Subproject: Preparation of an operational guide IEO-Vigo: Fumega de L´Hotellerie de Fallois, J.; to monitor marine pollution caused by a hydrocar- Nunes González, M.T.; Cabanas López, JM; bon oil spill and its impact on the resources. Castro Pampillón, JA.

START: July, 2004 RESEARCH SUPPORT: FINISH: December, 2006 Giraldez Rivero, R.

FINANCIAL ASSISTANCE: Ministry of Science and Technology within the Strategic Action on Maine Spills in the framework of the National R&D&I Program 2000-2003 (Special assistance: Strategic Action of R&D to combat acci- dental marine spills)

SUMMARY CONTINMAR is a project coordinated by CETMAR spills.Through subproject 3 “Preparation of an oper- focusing on the development of elements, tools, ational guide to monitor marine pollution caused by protocols for action and an information system for a hydrocarbon oil spill and its impact on the the design of contingency plans in the event of acci- resources”, collaboration was offered in the design dental marine spills. In keeping with the multidisci- of the databases. plinary character of the actions to be implemented to combat marine pollution, CONTINMAR comprises Under this subproject two reports were submitted 9 subprojects with the participation of a multidisci- on the oceanogrpahic characteristics of Galician plinary and interinstitutional group of researchers coastal waters and on the collections of existing with broad experience in the target areas of the oceanographic data. Currently being reviewed is a project in addition to experts belonging to the report on the characteristics of the spills and a man- administrations with competence for the formula- ual for the sampling and analysis of hydrocarbons. tion and execution of contingency plans. In 2005 work was carried out to prepare a sensitiv- Over the course of this year an information system ity map of the coast in which the entire Galician was implemented for the storage and management coast was classified on the basis of the sensitivity of the necessary information to be able to deal with scale (ESI) developed by the NOAA (Nacional emergency situations related to accidental marine Oceanic and Atmospheric Administration U.S.).

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Genetic evaluation of the phenotypical variants of García Vázquez, E. (Universidad de Oviedo). the European goose barnacle Pollicipes pollicipes across its geographic distribution. RESEARCHERS: CIMA: Molares Vila J. START: January, 2005 UO: Ayllón, F.; Campo Falgueras, D. FINISH: December, 2006 Centro Experimentación Pesquera: Fernández Rueda, P.; García Flórez, L. FINANCIAL ASSISTANCE: Local Ministry for Fisheries and Maritime Affairs RESEARCH SUPPORT: CIMA: Giráldez Rivero, R.; Centro Experimentación Pesquera: Cabranes Benduero, M.C. UO: González Pola, I.

SUMMARY In Spain and Portugal the goose barnacle The strategy used to determine the degree of (Pollicipes pollicipes) is harvested commercially from genetic differentiation of these biological units con- the intertidal populations, as it is one of the most sisted of examining their genetic variation for mark- highly esteemed shellfish. The edible part of the ers with varying degrees of preservation. Over the goose barnacle is the musculature found inside the course of 2005 this research focused on well-pre- peduncle, so specimens having the thickest peduncle served genes, which are usually the same or very fetch the highest prices in the market. Over the similar among different specimens belonging to the course of the past decade, the biomass and density same taxonomic unit. The study began with coding of goose barnacles of low commercial value –i.e. genes (loci) of the nuclear ribosomal RNA (rRNA), 5S individuals with a long peduncle and little muscula- rDNA y and the mitochondrial ones, 12S rDNA and ture- have undergone a substantial increase in many 16S rDNA. In order to conduct this study, it was nec- areas of the Galician coast. The increasing presence essary to devise the appropriate methodology, for of this type of goose barnacle, in detriment to the both DNA extraction as well as the amplification goose barnacle with a short, thick peduncle, has reactions (PCR) on the basis of oligonucleotide made it necessary to carry out a genetic characteri- primers that are heterologous (i.e. originating from zation of the populations harvested in order to another species). define the stock unit on which the effort of the goose barnacle fishery is exerted. The genetic markers studied with a higher level of variability were two mitochondrial sequences cod- P. pollicipes is the species inhabiting the east coast ing proteins: gene COI, which codifies subunit I of of the north Atlantic and is found on the coasts of the oxidase cytochrome enzyme and gene Cyt B France, Spain, Portugal, Morocco and Senegal. To which codifies cytochrome B. For these two genes, it date it is not known whether all of the goose barna- was also necessary to perfect the methodology used cles found on the coats of Europe and North Africa with this species. belong to a single stock or to several stocks with dif- ferent genetic characteristics. All the mitochondrial genes were subjected to complete gene sequencing and the identification of The basic question raised in this work is to deter- the genetic variants by means of the technique mine whether or not the two phenotypical variants called SSCP described by Sunnucks and coll. (2000) are based on a genetic determination or simply on was begun . With this technique it is possible to an environmental one. In 2005, the CIMA research detect variation on the sequence level in acryl amide unit conducted a detailed examination of the mor- gels in denatured conditions, causing the separate phological characteristics of the two variants, paying stands of DNA to migrate to a electrophoretic field. particular attention to the adult stage.

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Genetic diversity can also be studied in hypervari- barnacle samples, for four mitochondrial genes able loci (VNTR, such as, for example, the microsatel- (SSCP variation) and the preserved nuclear gene 5S lites) which allow for the identification of the stocks rDNA (variation in sizes of fragments amplified by (populations, subpopulations) as well as individuals means of PCR), although part of the latter requires from more diverse species, -vertebrates and inverte- the design of new primers to guarantee the brates. Using these types of genetic markers, several repeatability of the results. For the first time we suc- aspects of the biology of these species have been cessfully obtained complete sequences of these studied, ranging from reproductive strategies to genes and their variation has been analyzed in population structures. In the genus there has been goose barnacle samples from Asturias of the two no microsatellite marker published to date. In this phenotypes the “good” on and the “bad” one. The first year of the project heterologous microsatellites results obtained to date would point to the exis- (originating from another species) have been tested. tence of genetic differences between the two phe- However, in the end, they could not be used owing notypes of goose barnacle in two different mito- to their lack of specificity. chondrial genes for a sample from Asturias. A gene library is being compiled which will be completed by Over the course of 2005 we honed the method- the end of the project. ologies needed to analyze a large number of goose

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Byproducts from aquaculture in Galicia. Richie Flynn (Irish Farmer’s Association) Ireland

START: June, 2004 CHIEF RESEARCHER AT THE CIMA FINISH: December, 2005 Guerrero Valero, S.

FINANCIAL ASSISTANCE: RESEARCHERS: Aquareg INTERREG IIIC CIMA: Santos Piñeiro, I.; Outón Caamaño, M.J.

INTERNS: Otero Otero, M.

SUMMARY This research work is being carried out under a are the mussel, oyster and cockle among the mol- project financed by Interreg III delaing with byprod- lusks and the trout and turbot among the fishes. The ucts from aquaculture and includes the participation legislation passed by the European Union, National of Ireland, the Norwegian region of Trondelag and and Regional governments regulates and limits the Galicia. The objective is to compile a database with applications of the byproducts resulting from evis- the help of aquaculture waste producers, waste ceration, thereby avoiding the cannibalism that managers and companies that transform the waste, would occur if trout or turbot were given a diet of along with the applicable legislation on a regional, feed composed of offal from the same species. national and EU level. All th information collected Moreover, the new regulations open the door to during the execution of the project will be available new opportunities for the reuse and recycling of on the Internet. these materials.

Direct information was collected by grouping data The amount of waste from aquaculture in Galicia supplied by farmer’s associations through surveys comes to a total of 73,677 Tm./year not counting and indirect information was gathered through the the mud originating from feces from mussel culture bibliography available from the Local Ministry for which accounts for 237,491 Tm/year and, at the pres- Fisheries and Maritime Affairs of the Xunta de ent time, is not recycled. The next most abundant Galicia (regional Government of Galicia). The web waste are shells from the canning industry -53.456 page offers a database from Ireland, Norway and Tm/year- which are reused as different byproducts Galicia by menas of the following: by the companies Abonomar SL and Calizas Marinas SA and also included in the waste products is the - Preparation of Surveys. In keeping with the sur- debris generated form thinning out the culture vey model selected and used in the three countries, ropes, accounting for 6,854 Tm/year. visits were paid to farmer’s associations, aquaculture companies and waster managers. On the basis of the As far as the fishes are concerned, the total data obtained a list of farmers and managers was amount of waste produced from trout farming is drawn up to be included in the database. processed by the companies Toysal SA and Artabra SA, accounting for 4,036 Tm/year which can be bro- - Preparation of lists on the legislation applicable ken down into 2,700 Tm /year in mud recycled for to aquaculture waste as per the European Union, agricultural use and 907 Tm/year from evisceration. National and Regional governments. In turbot farms, the highest production of organic waste comes from dead fishes, for a total of 58 The species having the highest production –and Tm/year. therefore generating the greatest amount of waste-

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Powerpoint Presentations on the Internet: Presentation of the web page at the Irish Salmon www.mispeces.com (Spanish) and Growers´Asso. Conference, Bradán 2005. www.aquareg.com (English and Spanish). www.aquaculturebyproducts.com

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PROJECT STAFF

TITLE: CHIEF RESEARCHER: Carbon flows through Plankton in Subtropical Gutiérrez Morán, X.A. (IEO) Oligotrophic environments: a Lagrangian approach (CARPOS). INVESTIGADORES CIMA: Zapata Gago, M. START: December, 2003 UV: Fernández Suarez, E.; Marañón Sainz, E.; FINISH: December, 2006 Rosón Porto. G.; Serret Ituarte, P.; Teira González, E.; Varela Benvenuto, R. FINANCIAL ASSISTANCE: IEO: Bode Riestra, A.; de Armas Pérez, D.; Ministerio de Educación y Ciencia: González Estrada, N.; Nogueira, E.; CICYT REN2003-09532 Varela Rodríguez, M. UO: Anadón Álvarez, R.; Álvarez Marques, F

RESEARCH SUPPORT: Campaña Ferro, E.; Escánez Escánez; Espiñeira Piñeiro, M.; Lorenzo Salamanca, J.

INTERNS: UV: Aranguren Gassis, M.; Cermeño Ainsa, P.; Estévez Blanco, P.; González Fernández, J.; Fernández Graña, R.; Herrera Cortijo, J.; Pérez Rial, V.; Piedracoba Varela, S. UO: Blanco Bercial, L.; García López, E.; Paz Granda, A.

SUMMARY In recent years there has been much debate on The general objective of CARPOS is to quantify the magnitude of the net metabolic balance in the the net metabolic balance of carbon in the photic oligotrophic ocean, which boils down to the exis- layer of the central and peripheral region of the tence of two opposing points of view: respiration subtropical North Atlantic using a Lagrangian exceeds production in the oligotrophic ocean versus approach. This objective can be broken down into 4 the open ocean is in a state of metabolic balance. specific objectives: 1) verify that the central zone of the North Atlantic subtropical gyre has a net bal- The CARPOS project has come about largely from ance of the plankton community that is neutral or the results obtained by the applicant groups in pre- positive, 2) construct and compare the metabolic vious projects financed by the National Plan (CIR- balances in vitro and in situ in the comparison of CANA; CICYT MAR99-1072-01) or the European two ologotrophic regions, 3) quantify the effect of Union (CANIGO: MAS3-CT96-0060). The hypotheses small-scale physical variability on region’s produc- put forth by CARPOS are as follows: tivity and evaluate its effect on the net metabolic Hypothesis 1: The center of the North Atlantic balance and 4) assess the effect on methodological Subtropical Gyre has a compensated net metabolic limitations that may be associated with the execu- balance (close to 0). tion of in vitro experiments on the metabolic bal- Hypothesis 2: If the small-scale variability (inter- ance of this oceanic region. nal waves) is not considered, this will give rise to significant errors in the magnitudes of the estima- These objetives will be met through a work plan tions of the production and/or consumption of organized in 4 modules, which includes the execu- organic matter in the region. tion of 2 oceanographic surveys. The first survey Hypothesis 3: The heterotrophic metabolism char- (CARPOS-I) will carry out a Lagrangian sampling in acteristic of the subtropical NE Atlantic is, at least the central region of the North Atlantic subtropical partially, the result of methodological artifacts gyre, which is an area predicted to have a stable related to the maintenance of the microbial popla- metabolic balance. The second survey, CARPOS-II, tions enclosed in bottles. will focus its activity on a Lagrangian sampling in

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the peripheral region of the North Atlantic subtrop- The survey programmed for autumn 2005 was not ical gyre, known to present a clearly heterotrophic carried out due to a mechanical problem with the metabolic balance. oceanographic vessel shortly before we were sched- uled to set sail. If the necessary resources are avail- The task to be carried out at the CIMA (M. Zapata) able, the survey will be conducted in 2006. will consist of an analysis of the chlorophyll and carotenoids of the phytoplankton by means of HPLC. The data obtained will be processed using the CHEMTAX program, which will allow for the estima- tion of the contribution made by total chlorophyll a to the different taxonomic groups.

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CENTRO DE INVESTIGACIÓNS MARIÑAS CENTRO DE INVESTIGACIONS MARINAS (MARINE RESEARCH CENTER)

3.2. THESES AND RESEARCH WORKS (DOCTORAL PROGRAM) AND MASTER’S THESES CARRIED OUT AT THE CIMA

3.2.1. DOCTORAL THESES

TITLE: RESEARCHER: Evaluation of susceptibility to disease, immunologi- da Silva Escardua, P. M. cal capacity and productive traits of populations of the European flat oyster Ostrea edulis of diverse DIRECTOR: geographic origin. Villalba García, A.

DATE PRESENTED: July, 2005. AREA OF PATHOLOGY

SUMMARY Infection by Bonamia ostreae has been responsi- However, the variability between families within ble for high mortalities in the European flat oyster each origin caused some families to reach signifi- Ostrea edulis along the Atlantic coast of Europe cantly higher yields than others in their respective since the early 1980s. This disease is considered to be origins, which would imply that in a selection pro- a serious impediment to the development of the gram, it is not only the choice of origin, but also the European flat oyster industry in Galicia. The produc- choice of the pairs of parent stock that is important tion of a strain of oysters resistant to bonamiosis to production. may be a promising alternative. As a preliminary step to the development of a genetic selection pro- Several techniques using the optical microscope gram, it was considered of interest to evaluate the used to diagnose infection from B. ostreae, an possible existence of genetic variability in the popu- examination of histological cuts (CH), a smear of sev- lations of the European flat oyster for the traits eral of several organs monolayers of hemolymph under study and to choose a favorable population cells MCH) prepared using several protocols. The or populations on the basis of which a selection pro- MCH análisis was the most sensituve technique. It gram would be started. This report presents the was therefore chosen as the method used in all the results of the evaluation of the growth, mortality, samplings once the oysters had reached the appro- susceptibility to disease, immunological aspects and priate size for the extraction of the hemolymph. The gonadal development in oysters O. edulis from dif- results point to the need for a review of the recom- ferent geographic origins cultured in Galicia for 2 mendations of the Office Internationale des years (September, 2001 to September, 2003). Epizooties and the European Union for the diagno- sis of bonamiosis. A selection was made of genetically differentiated populations of oysters along the European coast – Infection by B. ostreae was the most serious dis- two areas free of bonamiosis, Greece (GR) eastern ease detected in oysters during the study period and Mediterranean and the North of Irlend (IR), and two caused mortality. Significant differences were found in Galicia, Ortigueira (OR) and Coroso (CO), with dif- in the prevalence of B. ostreae between origins and ferent histories of pressure from bonamiosis. Using between families within origins. Families with the parent stock taken from each of these populations, lowest and highest prevalence of infection by B. 5 families of oyster spat from each origin were pro- ostreae has the same origin (CO). The GR families, duced in the hatchery (with the exception of IR with the exception of one, exhibited a high suscep- which produced 4). The spat of the 19 families were tibility to bonamiosis. All the OR families manifested reared on a raft in the Ría de Arousa. Significant a lower susceptibility (low prevalence and intensity differences were found in growth, mortality and of the infection). Disseminated neoplasia was anoth- susceptibility to bonamiosis and other pathological er one of the pathological alterations having the alterations between origins and between families greatest prevalence. It was detected during almost within origins. The Galician oysters exhibited a high- all months and affected all the families, causing er yield in terms of production (more rapid growth mortality. An infection by the herpes virus was and higher survival). The Greek and Irish oysters, in another one of the diseases diagnosed and was contrast, showed low growth and survival rates. associated with mortality during the first year of cul-

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ture, with the IR origin being the most severely the plasma (PL) of the European flat oyster, and 14 affected. In addition, plasmodes and spores of the enzymes in HE and PL of Crassostrea gigas, which protozoon Haplosporidium armoricanum were was used as a reference, because of its resistance to observed, with a very low prevalencia and also seen bonamiosis. Significant qualitative and quantitative was an alteration characterized by lesions in the gills differences were found in the concentration of associated with a profusion of apoptosis, whose some enzymes between the origins of O. edulis and cause could not be identified. Other less important between O. edulis and C. gigas. The enzyme a-gly- symbionts and pathological alterations were detect- cosidase was the only one detected in the ed during the study, namely, spirochete bacteria, hemolymph of C. gigas que that was not detected in rickettsian-type colonies of organisms, trophozoites O. edulis. A comparison of the enzyme activity of the flagellate Hexamita nelsoni, ciliates and cope- between oysters that were infected and non-infect- pods. Unspecified histological symptoms of stress, ed by B. ostreae exhibited the presence of the such as hemocytic infiltration, granulocytomas and enzymes a-chemotripsine and a-glycosidase only in necrosis wee also observed. An index calculated on infected oysters and the concentration of other the basis of the prevalence of all the pathological enzymes was significantly higher in the hemolymph alterations observed estimated the total load of of infected oysters. The extra enzyme activity pathological problems (CTPP). The CTPP of the fam- detected in the hemolymph of the infected oysters ilies correlated significantly with the accumulated may be attributed to either the enzymes produced mortality at the end of grow-out. Significant differ- by the parasite or to the production of enzymes in ences were detected in the CTPP between origins the hemocytes induced by the infection, or a combi- and families within origins. Foreign origins present- nation of the two causes. ed the highest values, while OR had the lowest. The results obtained with the best families suggest that A study of the evolution of the gonadal condition the culture of the European flat oyster in Galicia is of the oysters over the course of grow-out on the possible if spat from suitable oysters are used. raft allowed us to determine two periods separated by a stage predominated by a gonadal rest period in Three types of hemocytes were distinguished in autumn 2002. In the first period, the oysters the hemolymph of the oysters: granulocytes (Gr), matured as males and in some oysters female game- large hyalinocytes (HIg) and small yalinocytes (HIp). togenesis developed later. Starting in autumn 2002, Also detected was a temporal variation in the total most of the oysters underwent female gametogene- number of hemocytes (THC) and in the relative sis and as of May 2003 the population was dominat- abundance of hemocytic types (DHC) of the ed by oysters carrying out male gametogenesis. hemolymph in the oysters, which would suggest a Significant differences were observed in the evolu- seasonal pattern. No significant differences were tion pattern of the gonadal condition between ori- found between THC and DHC between origins, gins. In the IR oysters the gonad was seen to be although the Galician oysters showed higher THC active in the most restricted period, and there was a values and percentage of Gr. as compared to the for- more widespread gonadal rest period. The gonadal eign specimens, which might indicate a higher activity of the GR oysters covered the broadest peri- immunological capacity. Significant differences were od, having a more restricted gonadal rest period recorded in the percentage of Gr and HIp between and the percentage of oysters that carried out families within origins IR and GR. A significant corre- female gametogenesis was smaller than in the other lation between the intensity of infection by B. origins. The two Galician origins exhibited a similar ostreae and THC and DHC was found, which, behavior, with a higher percentage of oysters depending on the progression of the infection will undergoing female gametogenesis in the second lead to an increase in the number of hemocytes cir- reproductive period. Significant differences were culating in the hemolymph (as well as those infiltrat- also detected between families within origins CO, ing the tissue), particularly the HIg which are GR and, to a lesser extent, IR, while the heterogene- assumes to be the type of hemocyte most favorable ity was lower in the OR origin. It was found that the to the division of the parasite. A trend was noted in probability of a B. ostreae infection reaching an which a high percentage of Gr was associated with advanced stage was significantly higher when the a higher survival rate and a lower susceptibility to oysters were in the pure female and predominantly disease. By The commercial kit api ZYM was used to female hermaphrodite sexual stage. Moreover, this dtect 16 enzymes in the hemocytes (HE) and 14 in probability was significantly higher when the oys-

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ters were in the stage of gonad maturity and after probably use up large quantities of energy, which the partial spawning. The development of gameto- would then be unavailable to confront an infection. genesis and, the production of oocytes, in particular,

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3.2.2. THESIS

TITLE: RESEARCHER: Spawning periods, larval abundance and recruit- Macho Rivero, G. ment of the cirripeds Balanus perforatus, Chthamalus montagui y Elminius modestus, in the DIRECTORS: south of Galicia (NW Spain). CIMA: Molares, J. U.V: Váquez, E. DATE PRESENTED: November 2005 AREA OF MARINE RESOURCES

SUMMARY Over the course of four years we have studied the Lastly we studied the effect of different environ- life history of three of the most important cirripeds mental factors (photoperiod, seawater and air tem- inhabiting the coasts of Galicia (NW Spain), and the perature, salinity and availability of food) on the impact of environmental factors on the different reproductive state, the presence of different larval stages of this cycle. In the open intertidal area of stages in the water column and recruitment in the Cabo Home (outer Ría de Vigo) there is an abun- intertidal zone. dance of the species Balanus perforatus and Chthamalus montagui, while in the more sheltered All of the species presented a very prolonged peri- zone of Pedras de Corón (inner Ría de Arousa) od of reproduction, larval abundance and recruit- Elminius modestus is the predominant species in the ment over the course of the year. A longitudinal pat- intertidal area, despite the fact that this species was tern was observed when we compared the data introduced in Europe a mere 60 years ago. from all the research carried out on the European coast, with a tendency towards broader periods as Of the three species studies in this work, there we move to the south or towards the have been no publications to date on the life histo- Mediterranean. ry of Balanus perforatus and Elminius modestus on the coast of the Iberian Peninsula, although some Moreover, we discovered different reproductive research has been done on the reproductive and strategies; Balanus perforatus and Elminius modes- recruitment periods of Chthamalus montagui. tus did not show synchronism at the onset or end of reproduction, although there did appear to be a cer- Over the course of the four year period of this tain degree of synchronism in the spawnings in the study (1998-2001), we determined the life history of middle of the reproductive period of B. perforatus. Balanus perforatus, Chthamalus montagui and In contrast, there was a strong degree of synchro- Elminius modestus, from fecundation to recruitment. nism at the onset and end of the reproductive peri- The first descriptive part of the analysis determines od in Chthamalus montagui, during which multiple the reproductive period, larval abundance and the asynchronic spawnings were carried out. recruitment period of the three species, and general patterns were obtained which describe the seasonali- A close spatial and temporal relationship was ty of the different stages of the life history of each found between the different larval stages under one. Next we focused on the influence of reproduc- study ( initial and advanced nauplii, cipris) both on tion on the larval abundance recorded in the medium an intra- and inter- specific level. Also, the presence as well as the impact of the previous two stages of of recruits in the intertidal zone is closely linked to the life history on recruitment. We also examined the the abundance in cipris and advanced nauplii, unlike interspecific relations in the three stages of the life what occurred during the reproductive period. history as well as the possible existence of spatial and temporal relations between larvae.

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The environmental factors did not appear to have tor does not mark the end of the reproductive sea- a major impact on the life history of Elminius mod- son in either of the two species, which would appear estus, possibly due to the extremely eurithermal and to depend on the availability of food as wells the eurihaline nature of this species in both the adult photoperiod in the case of C. montagui. and larval stages. None of the environmental factors studied deter- However, the reproduction of Balanus perforatus mine the larval abundance in this medium or the and Chthamalus montagui is controlled primarily by scope and intensity of recruitment of the three cir- seawater and air temperature, respectively, which ripeds on the Galician coasts. Hydrodynamic process- would appear to reflect the different positions the es yet to be studied in this work may likely be the two species occupy in the intertidal zone (lower factors having the greatest impact on these two intertidal zone for B. perforatus and the mid and stages of the life history of these species. upper zone for C. montagui). Nevertheless, this fac-

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3.2.3. ADVANCED STUDIES (DEA)

TITLE: RESEARCHER: Study of the reproductive cycle of the pullet carpet Cerviño Otero, A. shell clam Venerupis pullastra (Montagu, 1803) in two populations on the Galician coast: Camariñas DIRECTORS: and O Grove. CIMA: Martínez Patiño, D. USC: Abad Caeiro, M. DATE PRESENTED: July 2005 AREA OF AQUACULTURE

SUMMARY One of the most important species in the econom- the gonadal follicles have an abundance of mature ic activity of Galicia is the pullet carpet shell clam gametes, we also observed empty spaces left by (Venerupis pullastra) accounting for the greatest gametes already released and in the follicle walls volume of sales with the highest market price in new germinal lines appeared, which would explain recent years. Although the global income has the presence of mature gametes during the whole remained the same, catches have declined. To year. increase production, it is necessary to raise the issue of the rational management of the resource, on the Another important characteristic of this clam one hand, and the optimization of the hatchery cul- species is that is does not have a rest period, i.e., ture of this species. To this end it is necessary to have when the gonad stops producing gametes, it begins a thorough knowledge of the biology of this a new process of gametogenesis. The follicles, which species, paying close attention to reproductive still show signs of residual gametes, start to form aspects. vesicular cells which are filled, later to be invaded by new germinal lines that would give way to another This paper presents the preliminary conclusions cycle of gametogenesis. In Stage E4 of gonadal from the study carried out on two natural beds of renewal like in E1 of the onset of gametogenesis it the pullet carpet shell clam (Camariñas and O is still possible to find follicles with non-atresic resid- Grove), sampled monthly for fourteen months. In ual oocytes that may be released and new recruit- addition to gaining insight into the gametogenic ments may be generated. This has been confirmed cycle by means of traditional histological tech- in our lab, after obtaining spawnings of the pullet niques, we have studied the variation in the most carpet shell clam from these samplings all year important biochemical components of the gonad. round. For the analysis of glycogen content, the enzyme- based method of Keppler and Decker (1984) was As regards the analysis of the most important bio- used. Total proteins were obtained by the method chemical components in the gonad, we observed a of Lowry (1951) and for lipids Folch’s (1975) gravi- similar trend in both populations, with an increase metric method was used. in the quantity of proteins in the gonad during the spring, which also coincides with a decrease in the On the basis of a histological analysis of the sam- amount of lipids and glycogen during these same ples, it was deduced that both populations have a months. The decrease in glycogen and lipids coin- similar pattern of development despite the fact that cides with the months in which this species is in the they are subjected to different environmental fac- maturity and spawning stage, as can be seen from tors. The pullet carpet shell clam was assumed to the seasonal evolution in the Condition Index. have an extended period of maturity and spawning ranging from March to September in the Camariñas The wide-ranging maturity and spawning period population and during more months (although with (with continuous release and renewal of the a lesser proportion of individuals) in the O Grove gametes), the absence of a rest period and the fol- population. During this maturation period, in which licular asynchrony (which allows for the coexistence

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of spawning follicles with follicles accumulating found all year round and therefore, this species may reserves) are the three main characteristics that are potentially be able to produce spawnings in the peculiar to this species. Thanks to this, gametes are hatchery at any time of the year.

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TITLE: RESEARCHER: A study of disseminated neoplasia and other patho- Díaz Costas, S. logical alterations and parasites affecting popula- tions of the cockle, Cerastoderma edule, de Galicia. DIRECTORS: Carballal Durán, MJ.; Villalba García, A. DATE PRESENTED: July 2005 AREA OF PATHOLOGY

SUMMARY The cockle is a the most abundant mollusk of com- of surfaced individuals found on the beaches and mercial interest in the intertidal beds of Galicia, their greater vulnerability to predators, environ- which makes it an important source of income in the mental conditions and the sudden changes they are Galician economy. Despite the fact that it is a major subjected to in their intertidal habitat. The patho- economic resource, very few pathological studies logical alterations and parasites found to affect the have been carried out on the cockles population. cockle may be classified into three groups, in terms of the damage caused to the individuals. The most The objectives of this study are as follows: severe pathogenic alterations were disseminated neoplasia and large foci with strong hemocytic infil- Describe the pathological alterations affecting the tration and paratization by tremetodes. When the beds under study and to identify seasonality or rela- neoplasic cells were found in high intensities, they tionships with physical variables that might explain replaced the hemocytes and displaced the cockle tis- the tendency of the prevalence of the parasites and sue. The large foci of intense hemocytic infiltration pathological alterations to be able to design predic- mainly affect the foot, mantle gonad and digestive tion models that can be used in the harvesting plans system. These two pathological alterations were of this resource with a view to make better use of its associated with high mortalities of cockles on the harvest. north coast of Galicia. The following trematodes were observed: metacercarians located on the foot Compare the parasites and pathological alter- and mantle, and sporocysts. When sporocysts were ations as well as their intensity and prevalence in found in high intensities they caused gonadal cas- buried and surfaced cockles to determine their path- tration, and probably, the death of the individual. ogenicity. The metacercarians found in the foot could lead to problems when the cockle buries itself. The parasites Transmit disseminated neoplasia to healthy cock- that induced moderate lesions depending on the les by means of different techniques to demonstrate intensity of the infection were branchial bacteria, a possible viral origin. Pseudoklossia sp., the gregarine Nematopsis sp. and the branchial copepods. These parasites may pro- Pathological monitoring techniques are an impor- duce hemocytic reactions, which with high intensi- tant tool in establishing strategies to combat para- ties, would deform the tissue. The third group is sites and pathological alterations having a high made up of organisms had little or no pathological degree of pathogenicity. The design of models to effect, such as the undetermined gregarines, the predict prevalence and seasonality may be used in rickettsian type intracellular colonies, the turbellari- harvesting plans to make the most of the resource, um Paravortex cardii and ciliates. preventing the high mortalities owing to pathologi- cal anomalies. This research work characterizes the Several studies have shown that cockles that are parasites and pathological alterations found in com- enduring unfavorable conditions disinter them- mercially important natural beds of C. edule and selves. A comparison of the prevalence of the differ- discusses whether there is any seasonality or possi- ent parasites and pathological alterations between ble influence from physical factors such as tempera- buried and surfaced individuals exhibited signifi- ture and salinity in the appearance of these patholo- cant differences in prevalence and intensity of dis- gies. A study was also carried out to determine seminated neoplasia and large foci of hemocytic whether the parasites found in cockles may be the infiltration between the groups, which means that cause of their disinterment, due to the large number these two pathologies would appear to be the

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direct cause of the disinterment of the cockles. On studies that relate neoplasia to different types of the basis of histological preparations, it was found environmental pollutants: hydrocarbons, PCB, aro- that disseminated neoplasia and large foci of matic hydrocarbons, pesticides and cadmium, but intense hemocytic infiltration causes serious damage these works do not provide conclusive evidence that that could be lethal and possible affecting the sur- the above factors are the cause of disseminated neo- vival of the population owing to the castration that plasia. Several hypotheses have been put forth as to takes place in severely affected individuals. These the etiology of disseminated neoplasia. Although, pathological alterations may also bring about indirect none of these have yet to prove conclusive, there is mortalities, as the cockle, with a now weakened reason to believe that its origin in bivalves is caused immunological system, is left defenseless, on the sur- by an infectious etiology, probably a virus. In this face, subject to predators and sudden environmental research work, three transmission experiments were variations that are typical of the intertidal zone. carried out for the purpose of investigating the eti- ology of a possible infectious agent in disseminated The first cases of disseminated neoplasia in bivalve neoplasia in the cockle, Cerastoderma edule, in mollusks were described for Crassostrea virginica Galicia. In the first experiment the cohabitation of and Crassostrea gigas in North America. Today this healthy cockles with neoplasic specimens in the nat- pathology is found in 15 bivalve species. In cockles, ural environment was reproduced in tanks. This Cerastoderma edule, it has been found on the coast experiment confirmed the existence of a contagious of Brittany, France, in Ireland and in Galicia, north- agent. To determine the viral origin of the neopla- west Spain. Neoplasic cells in cockles are character- sia, two more specific experiments were conducted: ized by their large size and a higher proportion of the inoculation of neoplasic cells in healthy cockles, nucleus: cytoplasm than hemocytes. The nucleus is which would mean that the contagious agent wouls usually excentric, with a prominent nucleole, often have to be present in the cells and inoculation lobe-shaped. Mitotic shapes are also common. In the through the filtering of neoplasic cells where we initial stages of the disease it is manifested with the limited the size of the agent responsible for this dis- appearance of some abnormal cells in the conjun- ease to sizes of less than 0.45 mm, through which tive tissue around the gonad, digestive gland, man- protozoans, bacteria and neoplasic cells would not tle or gills. Neoplasic cell were also observed in the be able to pass. The transmission of neoplasia was cavities and vssels of the hemolymph. In more successful by cohabitation and the inoculation of advanced stages, most of the organs are affected neoplasic cells. However, transmission was not suc- where the normal cells are replaced by neoplasic cessful by filtering neoplasic cells, probably due to cells. The factor or factors that cause disseminated the short duration of the experiment because of the neoplasia have not been clearly defined. There are mortality of the inoculated cockles.

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3.2.4. MASTER’S THESIS

TITLE: RESEARCHER: Effect of feed containing carotenoids on the Fernandez Caballero, A.M. growth, coloration and composition of the blackspot sea bream. DIRECTORS: CIMA: Linares Cuerpo, F. MASTER:International University Master’s IEO_Vigo: Olmedo, M. in Aquaculture, ULPGC; ICCM; CIEHAM. AREA OF AQUACULTURE

SUMMARY This study constitutes the master’s thesis present- and 801.62±142.01 g and a mean size of 27.68±1.85 ed as a requirement for the IV Master Universitario cm and 27.82±1.78 cm respectively. The perivisceral Internacional en Acuicultura (2004–2006) (IV fat content in the fishes was 4% of the body weight International University Master’s course in for the two groups. Aquaculture) given on the Canary Islands from October 2004-March 2005. The study is based on Coloration was evaluated in three areas of the previous experiments carried out on farmed sparids body: the lateral zone (zone I), the caudal zone such as couch’s sea bream from the Canary Islands, (zone II) and the muscle (zone III) of the specimen. where feed containing carotenoids has been used to Samples were taken of the muscle, liver and gonads mitigate the great variations in the coloration of and an analysis was carried out to determine the cultured species as compared to those in the natural percentage of proteins, lipids, moisture, ash environment, a condition which makes these cul- (expressed in % of dry weight), the composition in tures fishes difficult to market. fatty acids and the concentration of carotenoids in the skin and muscle. The experiment was conducted over 4 months in experimental cages for sea bream belonging to The results showed that the use of the carotenoid Local Ministry of Fisheries and Maritime Affairs in dietary supplement in the blackspot sea bream did Bueu (Pontevedra) to evaluate the growth, bio- not produce any significant differences in growth in chemical composition and coloration of the sea either group of fishes. Nor did it improve the bio- bream specimens. Two types of feed were used, one chemical composition of the muscle and liver of the having a composition of 50/18 and supplemented specimens. with carotenoids, a diet of 40mg/Kg based on shrimp flour and Krill (B-carot) and the other one, As regards coloration, a significant difference was used as a control, SORGAL 50/12 (S-12). Five batches found in the Hue (color) of the skin (zone I) at the of 900 fishes each were used (Pm 192.75±30.18 g time of butchering and in the Chroma (intensity) of and Tm 17.96±1.47), 3 of which were fed a diet of B- the muscle (zone III) in the smaller-sized fishes and carot and 2 with S-12. in the L (lightness) of the skin (zone I) at the time of butchering and in the Chroma (intensity) of the Another two batches of 500 fishes hatched in muscle (zone III) of the larger sized-fishes, in a com- 2001 (Pm 664.55±111.12 g and Tm 25.73±2.17) were parison of the groups of fishes given feed supple- introduced in two modules each. One batch was fed mented with carotenoids and the control groups. B-carot and the other received S-12. Three sam- plings were conducted, - at the beginning of the A comparison of the fishes given the above treat- experiment, and 61 and 118 days after starting to ment with specimens in the natural environment, administer the pigmented diet. In December 2005 led to the conclusion that a diet with a carotenoid the fishes fed the two types of feed did not show supplement (40 mg/Kg diet) administered over a any differences in growth. The fishes fed S-12 and B- period of 118 did not produce the desired effect. carot attained a mean weight of 789.39±125.50 g Significant differences were found between fishes

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fed the pigmented diet and those from the natural stage, we would suggest that this study be conduct- environment in all three values L (lightness), Hue ed over a longer period of time to be able to draw (color) and Chroma (intensity). definitive conclusions on the advisability of the use of feed containing carotenoids in the grow-out Since this experiment was only carried out over a stage of the blackspot sea bream. 4 month period and taking into account that the blackspot sea bream at this point is in a slow growth

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TITLE: RESEARCHER: Growth and reproduction of the Pacific oyster, Escudeiro Rossignoli, A. Crassostrea gigas (Thunberg, 1793), cultured in intertidal zones and on rafts in Galicia (NW Spain). DIRECTORS: CIMA: Guerra Diaz, A. MASTER. Univ. do Algarve, Portugal: Dinis, Mª.T. International Master’s in Aquaculture and Fisheries. Univ. do Algarve. (Portugal). AREA DE AQUACULTURE

SUMMARY Oyster production in Galicia is based primarily on 1793), cultured in intertidal zones and on rafts in the rearing of spat of the European flat oyster, Galicia (NW Spain)” is included within the above Ostrea edulis, imported from third countries such as project and its activity is focused on monitoring the France, Italy and Ireland, despite the fact that this growth pattern and reproductive cycle of two culture has always entailed problems with mortali- batches of adult Crassostrea gigas 18 months old ties and a lack of spat supplies. For this reason, originating from the Bay of Arcachon (France). Two among the alternatives put forth to aid the oyster different culture techniques were used: the elevated sector, the Galician Administration has introduced method using “pochones” (bags) in the intertidal the idea of a supplementary culture of the species zone and suspended from rafts located at two Crassostrea gigas, known in Galicia as the “ostra points in the Ría de Arousa (Galicia), Vilaxoán and rizada” (Pacific oyster) which does not appear to Grove F. Monthly histological studies on the gonads present any major reproductive problems. However, of around 15 - 30 individuals from each batch owing to the fact that this species is allochthonous, showed that summer spawnings occurred in both it has generated a great deal of mistrust. Along study locations, coinciding with the warmest months these lines, the Directorate General of Marine of the year. However, these clutches were not viable, Resources started a project aimed at evaluating the since the attachment of juvenile oysters did not take feasibility of farming the Pacific oyster in Galicia place. However, the correlation coefficients from an integral standpoint –including technical, between the gonadal maturity indices and physio- biological, economic and ecological aspects- as a logical conditions were statistically significant in possible supplement to the culture of the European both batches. In general, the results obtained on the flat oyster. biometric variables (dorsal-ventral length, mean total weight and fresh meat weight) showed that The study entitled “Growth and reproduction of growth was slightly higher in the intertidal zone the Pacific oyster, Crassostrea gigas (Thunberg, than on the rafts.

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SCIENTIFIC ACTIVITY

3.3. ADVISORY SERVICES FOR THE SECTOR AND THE ADMINISTRATION

A.- Reports issued.

- Report on the pathology of populations of the coc- - Report on the pathological condition of a sample kle, Cerastoderma edule, and carpet shell clam, of carpet shell clam received on 9/09/2005. Ruditapes decussatus, of the Lombos de Ulla zone, Requested by the Fishermen’s Association of under the “Action Plan for the Recovery of Lombos Redondela owing to the presence of an incident of do Ulla”. DXRM. Carballal Durán M.J.; Fernández mortality (70-80%) on A Portela beach (Cesantes). Conchas, R.; Villalba García, A. DXIDP. López Gómez, C.

- Weekly reports issued (43) in 2005 on the larval - Report on the mortality of the fauna inhabiting abundance of the mussel for the CCMM and INTEC- the rocky areas on the Island of Ons observed MAR. Fuentes González, J.M. during the summer of 2005. Delegación Territorial de Vigo of the CPAM. Molares Vila, J. - Report on the culture of Haliotis discus hannai in Galicia. DXRM. Guerra Díaz, A. - Project Report on the promotion of the culture of new species of sparids: blackspot sea bream. - Report on the emission of wastewater from a mini- Pilot trials and technological transfer JACUMAR. hatchery on bivalve mollusks located in the munici- Linares Cuerpo, F. pality of O Vicedo (Lugo). DXRM. Guerra Díaz, A. - Annual Report on the culture of the blackspot - Reuse of effluents from a seawater fish farm for sea bram in cages in Bueu, of the Local Ministry the grow-out of bivalve mollusk spat. Final report of for Fisheries. DXRM. Linares Cuerpo, F. research project PGDIT. IDP. Guerra Díaz, A.

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B.- Participation in scientific or technical meetings.

- Manager of the Marine Resource Program of - Representative of the Galician Regional the Galician Plan, IDIT. Blanco Pérez, J.C. Government Gallega in the Working group of the National Plan, JACUMAR “Promotion of the - Member of the Organizing Committee X culture of new species of sparids. Pilot trials and Congreso Nacional de Acuiculture. Gandía technological transfer”. Linares Cuerpo, F. (Valencia). Cerviño Eiroa, A. - Member of the working group at the - Member of the Organizing Committee IV International Seminar on “Octopus culture”. Symposium on Sandy Beaches 2006. Sánchez Linares Cuerpo, F. Mata, A. - Member of the working group of the National - Member of the Organizing Committee of the I Plan, JACUMAR “Sole culture”. Linares Cuerpo, F. Seminario de Gestión y Tratamiento de Residuos en la Acuicultura. Vigo. (Pontevedra). Guerrero - Working group on “Fishery Research in Spain” Valero, S. for the preparation of the Libro Blanco de la Pesca. Ministerio de Agricultura, Pesca y - Member of the Organizing Committee of the Alimentación. Vigo. Guerra Díaz, A.; Molares VII Foro dos Recursos Mariños. O Grove Vila, J. (Pontevedra). Guerra Díaz, A. - Participant at round table on “A Pesca ¿Unha - Participation as technical advisor of the Local Actividade Sostible?”. University of Vigo. Ministry for Fisheries and Maritime Affairs at Molares Vila, J. two meeting to coordinate the campaign to supply mussel spat. CIMA Vilanova de Arousa. - Meeting at the General Fisheries department Fuentes González, J.M. on the program for the recovery of hake and the Norway lobster of the Cantabrian Sea and the - Participation in the Round Table. Libro Blanco Western Iberian Peninsula. Molares Vila, J. “Plan de Galicia do Mar”. Aquaculture. Guerra Díaz, A. - Working group on deep sea fishery for the pre- paration of the Libro Blanco de Política - Representative of the Regional Government of Marítima de Galicia. CPAM. Molares Vila, J. Galicia in the monitoring group of the National Plans JACUMAR. Guerra Díaz, A. - Working group on research in the world of fis- hery. Libro Blanco de Política Marítima de - Meeting of the WGMASC of the ICES Galicia. CPAM.. Molares Vila, J. Seafarming Committee, participant as a member of the WGMASC. La Rochelle (France). Fuentes - Bilateral meeting on fishery research carried González, J.M. out in Spain and Canada. St John’s (Canada). Canadian Embassy. Molares Vila, J. - Member of the Galician Scientific Fishery Committee (CCGP). Participant in two meetings - Working Group on “Los Caladeros del Golfo de of the CCGP. Fuentes González, J.M. Cádiz y Portugal” for the preparation of the Libro Blanco de la Pesca. Ministerio de - Member of the Galician Scientific Fishery Agricultura, Pesca y Alimentación. Sevilla. Committee (CCGP). Participant in two meetings Molares Vila, J. of the CCGP. Participant in drawing up a report on the condition of fishery stocks of interest to - Participant at the ASLO International Meeting. Galicia. Molares Vila, J. Santiago de Compostela. Zapata, M.

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- Meetings for the coordination and preparation - Meeting for the coordination of a working of the final report of the participants in the pro- group of participants in AQUAREG. Brussels. ject “ Bonamia ostreae life cycle investigations, Guerrero Valero, S. optimised production of resistant Ostrea edulis spat, and studies of oyster (Ostrea edulis) immu- - Meeting for the coordination of the project, ne mechanisms”. Cork, Irleland and London, Sustainable Harvesting of Ensis (SHARE-90). U.K. Villalba, A. Faro-Portugal. Da Costa González, F.; Martínez Patiño, D. - Meeting for the coordination of the working - Meeting INTERREG IIIB: “Sustainable harves- group of Project AQUAREG. I. Communication. ting of Ensis (SHARE-90)”. Ireland. Martínez Waste and byproducts from aquaculture. Patiño, D.; Da Costa González, F Villagarcía de Arousa. Guerrero Valero, S.

- Meeting in London, U.K., with “BY-PRODUCTS, Working Group of UK. Guerrero Valero, S.

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C.- Other

- Member of the panel of judges to award the b) proteins, lipids and fatty acids in larva samples of Diploma of Advanced Studies to Seila María Díaz gilt head sea bream from cultures carried out at the Costa and Ana Cerviño Otero. Carballal Durán, M.J.; IEO in Mazarrón (Murcia). Guerra Diaz, A. c) proteins, lipids and fatty acids in samples of larvae and fry of the sole (Solea senegalensis) from cultures - Member of the WGMASC of the ICES seafarming carried out in the culture centers at the IEO in Committee. Fuentes González, J.M. Santander. proteins, lipids and fatty acids in samples of paralar- - Atenndance at IFAS Aquaculture conferences and vae of the octopus and prey from cultures carried visit to the BMW zone of Ireland in the framework out at the CO in Vigo of the IEO of the project RE-GEX of AQUAREG ITERREG IIIC. Darriba Couñago, S.; Martínez Patiño, D.; Ojea - Visit to the BMW region of Ireland. Project RegEx Martínez, J. of AquaReg. Abalone culture facilities. Darriba Couñago, S.; Matínez Patiño, D.; Ojea Martínez, - Irish Shellfish Association annual conference. Justa. Clarinbride.Co. Galway. Ireland. Darriba Couñago, S. - Visit to the BMW region of Ireland. Project RegEx of AquaReg. Facilities of the Marine Institute in - I Forum AquaReg. Vigo. Pontevedra. Darriba Galway and Newpor. Cerviño Otero, A.; López Couñago, S. Gómez, C.; Guerra Díaz, A.; Darriba Couñago, S.

- Collaboration in the organization of the I - Visit to the Trondelag region (Norway). Project Seminario de Gestión y Tratamiento de Residuos en Regex ofe AquaReg. Sea urchin culture facilities. Da la Acuicultura. Otero Otero, M. Costa González, F.; Loureiro Barcon J.M.; Ojea Martínez, J. arriba Couñago, S.; Matínez Patiño, D.; - Analysis and attendance –non ordinary- at the fish Ojea Martínez, J. culture lab. Linares, F.: a) fatty acids in phytoplankton samples from the company TinaMenor, carried out in the fish cul- ture lab.

114 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

3.4. SCIENTIFIC AND TRAINING ACTIVITIES CARRIED OUT BY THE STAFF

3.4.1. ORGANIZATION, CHAIRMANSHIP AND PARTICIPATION IN SCIENTIFIC MEETINGS

VII Foro dos Recursos Mariños. O Grove X Congreso Nacional de Acuicultura. Gandía (Pontevedra). (Valencia).

Member of the Organizing Committee. Guerra Cerviño Eiroa, A.; García Fernández, A.; De Coo Díaz, A. Martín, A. Grow-out of the short-neck clam, Ruditapes philippinarum (Adams & Reeve, 1850), in Cerviño Eiroa, A.; García Fernández, A.; De Coo different culture system sin the Rías Bajas and on the Martín, A. method using bags for the grow-out of coast of Galicia. Book of abstracts: 600-601. clams on rafts. Cerviño Eiroa, A.; García Fernández, A.; De Coo Cerviño Otero, A.; Ojea, J.; Nóvoa, S.; Martínez, D. Martín, A. Results of the grow-out of the carpet Reproductive cycle of the pullet carpet shell clam shell clam, Ruditapes decussatus (Linné, 1758), using Venerupis pullastra (Montagu, 1803). different systems in the Rías Bajas of Galicia. Book of abstracts: 598-599. Da Costa, F.; Ojea, J.; Nóvoa, S.; Martínez, D. Culture of hatchery-reared spat of three commercial sole- Cerviño Otero, A.; Ojea, J.; Nóvoa, S.; da Costa, F.; nacean species of Galicia. Martínez, D. Study of the reproductive cycle of the pullet carpet shell clam Venerupis pullastra Darriba, S.; Miranda, M. Impact of declining salinity (Montagu, 1803), in two different populations on on the reproduction of the razor clam (Ensis arcua- the Galician coast: Camariñas (NW Galicia) and O tus). Grove (SW Galicia). Book of abstracts: 508-509.

Guerrero, S.; Santos, I.; Outón, M.J. Characterization Cerviño Otero, A.; Martínez, D.; Nóvoa, S.; Ojea, J.; and quantification of waste from aquaculture in García-Martínez, M.; Abad, M. Seasonal variation in Galicia. the condition index and biochemical composition of the gonad of Venerupis pullastra (Montagu, 1803) in Miranda, M.; Santamaría, I.; Casal, J.; Ruiz, A.; two different populations on the Galician coast. Guerra, A. Use of wastewater from a turbot fish Book of abstracts: 606-607. farm in Galicia for the grow-out of bivalve mollusks. Cerviño Otero, A.; Carballal, M.J.; Martínez, D.; Rodríguez, L.; Iglesias, D.; Escudeiro, A.; Montes, J.; García Fernández, A.; Cerviño Eiroa, A. Effects of Conchas, R.F.; Pérez, J.L.; Fernández, M.; Guerra, A. processes related to the first reproductive maturity Initiative to promote the culture of the Pacific oyster in the grow-out of the clams, Ruditapes decussatus (Crassostrea gigas) in various Galician rías. (Linné, 1758), Venerupis pullastra (Montagu, 1803) and Ruditapes philippinarum (Adams & Reeve, I Seminario de Gestión y Tratamiento de Residuos 1850). Book of abstracts: 604-605. en la Acuicultura. Vigo. (Pontevedra). Cerviño Otero, A.; Carballal, M.J.; Martínez, D.; Guerrero Valero, S. Waste from aquaculture in Cerviño Eiroa, A.; García Fernández, A. Differential Galicia. growth between males and females of Venerupis pullastra (Montagu, 1803) and the repercussions on practices in hatcheries and grow-out centers. Book of abstracts: 602-603.

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Chereguini, O.; Garcia de la Banda, I.; Lobo, C.; Mariño- Balsa,J.C., F.Linares and J.L.Rodríguez. Linares, F. Larval co-feeding and weaning of Solea Design of a plan to restock the turbot in Galicia. senegalensis (Kaup, 1958) in environments with dif- Book of abstracts:128-130 ferent salinities. : 366-367. Olmedo, M.; Linares, F.; Álvarez-Blázquez, B.; Da Costa, F.; Darriba, S.; Martínez, D. Embryonic and Peleteiro, J.B. Current state of the blñackspot sea larval development of Ensis arcuatus (Jeffreys, 1865) bream, Pagellus bogaraveo. Book of abstracts: 356- using the electron scanning microscope. Book of 357. abstracts L: 610-611. Ojea, J.; Martínez, D.; Nóvoa, S.; Cerviño-Otero, A. Da Costa, F.; Nóvoa, S.; Ojea, J.; Martínez, D. Gonadal cycle of a population of Tapes philip- Conditioning of the solenacean Ensis arcuatus under pinarum and biochemical composition (glycogen, two temperature regimes: correlation to proteins lipids and proteins) dof the gonad-visceral mass. and glycogen. Book of abstracts: 608-609. Book of abstracts:638-639.

Darriba, S.; Miranda, M.; Guerra, A. Reproductive Prado, S.; Ojea, J.; Martínez, D.; Montes, J.; Barja, J. study of the bivalves Ensis arcuatus and Ensis siliqua Selection of marine bacteria as probiots in aquacul- in different natural beds in Galicia (NW Spain). ture. Activity of the strain. Book of abstracts 244- Book of abstracts: 510-511. 245.

De Coo, A.; Cerviño Eiroa, A.; García Fernández, A. Rodríguez, L.; Iglesias, D.; Montes, J.; Conchas, R.F.; Critical period of mortality in the grow-out of the Pérez, J.L.; Fernández, M.; Guerra, A. Study of the European flat oyster Ostrea edulis (Linné, 1758) on evaluation of the culture of the Pacific oyster rafts in the Rías Bajas of Galicia. Correlation to cul- (Crassostrea gigas) in various Galician rías. ture densities and handling operations. Book of Preliminary results. Book of abstracts 578-579. abstracts: 612-613. Santos, I.; Guerrero, S.; Outón, M.J. Waste from tur- García Fernández, A.; Cerviño Eiroa, A.; De Coo bot culture (Psetta maxima) in Galicia. Book of Martín, A. Growth Potential and mortality results in abstracts: 138-139. the grow-out of Venerupis pullastra (Montagu, 1803), using several methods in the Rías Bajas of IX Congreso Ibérico de Parasitología. Coimbra Galicia, with special attention being paid to the fac- (Portugal). tors interacting with its evolution. Book of abstracts: 618-619. Conchas, R.F.; Pérez, J.L.; González, G.; Montes, J. Optimization of the culture of the European flat Guerra, A. Achievements in the study of commercial oyster in Galicia through the control of the parasite solenaceans in Galicia, under the National Plan Bonamia ostreae using quick diagnostic techniques. JACUMAR. Book of abstracts: 574-575. López, C.; Darriba, S.; Conchas, R.F. Presencia de Guerrero, S.; Santos, I.; Outón, M.J. Use of byprod- Marteilia sp. in Solen marginatus (Pennántt, 1777) ucts from bivalve shells in Galicia. Book of abstracts: en Galicia (NO España). 120-121. 40 EMBS: European Marine Biology Symposium. López, C.; Darriba, S.; Miranda, M.; Álvarez, C. Vienna. Depuration of Ensis arcuatus and Ensis siliqua (Solenacea). Book of abstracts: 624-625. Sánchez Mata, A. Macrobenthic biomass gradient over a range of exposed sandy beaches at the north- Miranda, M.; Santamaría, I.; Casal, J.; Ruiz, A.; ern Atlantic coast of Spain. Guerra, A. Grow-out of clam spat in effluents from land-based marine fish farms. Book of abstracts: Sánchez Mata, A. Patterns of zonation in Antarctic 630-631. intertidal benthos: the effects of glacial melting on diversity and abundance.

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Fifth International Conference on Molluscan Aquaculture Europe 2005. Trondheim Shellfish Safety. Galway (Ireland) (Noruega).

Blanco Pérez, J.C. Modelling phycotoxins accumula- Guerrero, S.; Santos, I.; Outón, M.J. Mussel and tion in bivalves: a review. white shell by-products in Galicia, Spain.

Blanco, J.; García, A.; Vázquez, M.C.; Mariño, M.C.; Santos, I.; Guerrero, S.; Outón, M.J. Turbot Farms Gómez, M.J.; Cano, J.; Campos, M.J. Kinetics and waste in Galicia, Spain. anatomical distribution of Paralytic Shellfish Poisoning (PSP) toxins in the king scallop Pecten 12th International Conference of the European maximus. Association of Fish Pathologists “Diseases of Fish and Shellfish”. Copenhagen (Denmark). García-Muñoz, A.; Campos, M.J.; Cano, J.; Mariño, C.; Gómez, M.J.; Blanco, J. Anatomical distribution Conchas, R.F.; Balseiro, P.; Novoa, B.; Montes, J.; of amnesic shellfish poisoning (ASP) toxins in the Figueras, A. Validation of nested PCR for diagnosis king scallop Pecten maximus throughout the culture of parasite Perkinsus olseni/atlanticus in culture cycle. clams.

Saavedra, Y.; González, A.; Fernández, P.; Blanco, J. The influence of sex and spawning on metal concen- Carballal, M.J.; Díaz, S.; Villalba, A. Urosporidium sp. tration and content of Mytilus galloprovincialis. hyperparasite of the turbellarian Paravortex cardii in the cockle Cerastoderma edule. Larvi 2005. 4th Fish & Shellfish Larviculture Symposium. Ghent University, Belgium. Díaz, S.; Soudant, P.; da Silva, P.M.; Carballal, M.J.; Lambert, C.; Villalba, A. Flow cytometric DNA con- Laura Ribeiro, L; Couto, A.; Olmedo, M.; Álvarez- tent analysis of normal haemocytes and neoplastic Blázquez B.; Linares, F.; Valente, L. Digestive cells in the haemolymph of the cockle Cerastoderma enzymes activity during black spot seabream edule. (Pagellus bogaraveo) larvae and post-larvae devel- opment. III Congreso Internacional de Ciencia y Tecnología Marina “Océanos III Milenio”. Puerto Real (Cádiz).

Marine and Freshwater Toxins Analysis. First Joint Díaz, S.; Carballal, M.J.; Villalba, A. Alteraciones Symposium and AOAC Task Force Meeting. Baiona patológicas del berberecho, Cerastoderma edule, de (España). un banco natural del NO de España.

Blanco, J.; Mariño, M.C.; Martín, H.; Acosta, C.P. Vibrio 2005. Gante (Bélgica). Anatomical distribution of DSP in the mussel Mytilus galloprovincialis during the depuration process. Prado, S.; Romalde, J.L.; Montes, J.; Barja. J.L. New Vibrios isolated from disease outbreaks in shellfish World Aquaculture 2005. Bali (Indonesia). larvae.

Chereguini, O.; García de la Banda, I.; Diez, J.; Lobo, National Shellfisheries Association Annual Meeting, C.; Linares, F. Effect of different regimes on Senegal Philadelphia (PN. EEUU). sole Solea senegalensis (Kaup 1858) larval culture in Cantabria (Northern Spain). Da Silva, P. M.; Fuentes, J.; Villalba, A. Disseminated neoplasia in oysters Ostrea edulis cultured in Galicia García de la Banda, I.; Chereguini, O.; Lobo, C.; Diez, (NW Spain). J.; Linares, F.; Arce, F. Ongrowing of senegal sole Solea senegalensis (Kaup 1858) fries using two Da Silva, P. M.; Villalba, A.; Sunila, I. Branchial lesions different commercial diets in Cantabria (Northern associated with abundant apoptotic cells in oysters Spain). Ostrea edulis of Galicia (NW Spain).

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La Peyre, J.; Casas, S.M.; Reece, K.S.; Villalba, A. In International Moolusk Culture Workshop on “The vitro propagation of Perkinsus mediterraneus a par- current state of the biology and culture of byruning asite of the edible oyster Ostrea edulis. bivalves”. Universidad de Concepción. Chile.

Romalde, J. L.; Rodríguez, J.M.; Beaz, R.; Vilariño, Guerra, A.; Darrriba, S. New species in the aquacul- M.L.; Carballal, M.J.; Díaz, S.; Villalba, A. Evidences ture of bivalve mollusks: studies on solenaceans in for retroviral etiology of the disseminated neoplasia Galicia (Spain). in cockles (Cerastoderma edule). Guerra, A. Mini-hatcheries: development and trans- fer of a facilities model for the procurement of com- mercial bivalve mollusk spat.

Ojea , J. Culture of microalgae to feed burying bivalve mollusks.

Martínez, D. Cultures of three solenacean species of commercial interest in Galicia.

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3.4.2. DIRECTION OF DOCTORAL THESES AND INTERN TRAINING

• Post-Graduate Degrees

At the CIMA

A.- Doctoral Theses Subject: Larval ecology of the goose barnacle Pollicipes pollicipes: seasonal patterns, control Subject: Depuration of ASP toxins in the great scal- mechnaisms and behavior from eclosion to attach- lop. ment. Doctoral Candidate: Mauríz Pereira, A. Doctoral Candidate: Macho Rivero, G. Degree: Master’s Degree in Marine Science Degree: Master’s Degree in Marine Science Tutor and thesis director: Blanco Pérez, J.C. Tutor and thesis co-director: Molares Vila, J.

Subject: Pathological study of the cockle population Subject: Harvesting the cockle Cerastoderma edule Cerastoderma edule (L.) in Galicia. in Galicia. Biology, ecology and management of the Doctoral Candidate: Iglesias Estepa, D. natural beds. Degree: Master’s Degree in Marine Science Doctoral Candidate: Santos Piñeiro, I. Tutor and thesis director: Carballal Durán, M.J.; Degree: Master’s Degree in Biology Villalba García, A. Tutor and thesis director: Molares Vila, J.

Subject: Study of disseminated neoplasia and other Subject: Evaluation of the susceptibility to disease, pathological alterations and parasites affected the immunological ability and productive traits in popu- populations fo the cockle Cerastoderma edule in lations of the European flat oyster Ostrea edulis of Galicia. different geographic origin. Doctoral Candidate: Díaz Costa, S.M. Doctoral Candidate: da Silva Scardua, P.M. Degree: Master’s Degree in Marine Science Degree: Master’s Degree in Biology Tutor and thesis director: Carballal Durán, M.J.; Tutor and thesis director: Villalba García, A. Villalba García, A. Subject: Comparison of immunological parameters Subject: Biochemical and immunological identifica- between oyster varieties (Ostrea edulis) and species tion of mussel larvae, Mytilus galloprovincialis. (O. edulis and Crassostrea gigas). Doctoral Candidate: Lorenzo Abalde, S. Doctoral Candidate: Comesaña Lestayo, P. Degree: Master’s Degree in Biology Degree: Master’s Degree in Marine Science Tutor and thesis director: Fuentes González, J.M. Tutor and thesis director: Villalba García, A.

Subject: High quality hatchery reared mussel spat . At other institutions Doctoral Candidate: Domínguez Pérez, L. Degree: Master’s Degree in Biology Subject: Accumulation of metals in bivalves. Tutor and thesis director: Fuentes González, J.M. Doctoral Candidate: Saavedra Yáñez, Y. Degree: Master’s Degree in Chemistry Subject: Optimization of the culture of the pullet Institution : INTECMAR carpet shell clam (Venerupis pullastra). Tutor and thesis director: Blanco Pérez, J.C. Doctoral Candidate: Cerviño Otero, A. Degree: Master’s Degree in Biology Subject: DSP toxins in phytoplankton and bivalve Tutor and thesis director: Martínez Patiño, D. mollusks. Doctoral Candidate: Fernández Cañamero, M.L. Degree: Master’s Degree in Chemistry Institution: CETMAR Tutor and thesis director: Blanco Pérez, J.C.

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Subject: Inmunodetection of mussel larvae on the Subject: Effects of the black tide from the Prestige Galician coast by means of monoclonal antibodies M oil spill on intertidal benthos of the open-ocean 22.8 and M 36.5. beaches of Galicia. Doctoral Candidate: Pérez Estévez, D. Doctoral Candidate: Sánchez-Mata Arabella G. Degree: Master’s Degree in Biology Degree: Master’s Degree in Biology Institution: Department of Immunology, School of Institution: University of Vigo Sciences. University of Vigo Tutor and thesis director: Sánchez Mata, A. Tutor and thesis director: Fuentes González, J.M. B.- Practical internships for Master’s Degrees at the Subject: Bonamia sp. histopathological alterations CIMA of Ostrea puelchana d´Orbigny (Mollusca, Pelecypoda, Ostreidae), from the natural beds and Graduate.: Escudeiro Russignoli, A. culture areas of the Gulf of San Matías, Argentina”. Degree: Master’s Degree in Biology Doctoral Candidate: Kroeck Sesgonds, M. Tutor: Guerra Díaz, A. Degree: Master’s Degree in Biology Institution: Instituto de Biología Marina y Pesquera Graduate.: Fernández Caballero, A.M “Almirante Storni” (Argentina) Degree: Master’s Degree in Marine Science Tutor and thesis director: Montes Pérez, J. Tutors: Linares Cuerpo, F.; Olmedo, M.

Subject: Study of the production of spat of the European flat oyster (Ostrea edulis L.) by emans of microbiot modification. Doctoral Candidate: Prado Plana, S. Degree: Master’s Degree in Biology Institution: University of Santiago de Compostela Tutor and thesis director: Montes Pérez, J. and Barja Pérez, J.L.

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• VOCATIONAL INTERNS

Tutor: Carballal Durán, M.J. Tutor: Martínez Patiño, D. Intern: Angueira Souto, M.C. Intern: Rial Conde, D. Specialist in Pathology Specialist in Aquatic Production

Tutor: Guerrero Valero, S. Tutor: Molares Vila, J. Intern: Otero Otero, M. Intern: Crego Mata, A. Specialist in Aquatic Production Systems specialist

Tutor: Linares Cuerpo, F. Tutor: Molares Vila, J. Intern: Soto Fraga, N. Intern: Varela Sóñora, M.T. Specialist in Aquatic Production Specialist in Aquatic Production

Tutor: Linares Cuerpo, F. Intern: Cachafeiro Diéguez, P. Specialist in Aquatic Production

Tutor: Martínez Patiño, D. Intern: Rábade Uberos, S. Specialist in Aquatic Production

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• On-the-job Training (FCT).

A.- University degrees B.- Technicians (Voctional training)

Student: González Méndez, I. Student: Goldar Buceta, S. From: University of Vigo From: I.E.S. “A Xunqueira”. Pontevedra Period: August - 05-February 06 Period: January-June Tutor: Martínez Patiño, D. Tutor: Carballal Durán, M.J.; Montes Pérez, J.

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3.4.3. COURSES GIVEN

DOCTORAL PROGRAMS

Marine Biology and Aquaculture Program. Summer Course. University of Oviedo

Course: Episodes of phycotoxicity in mollusks . Fundamentals of Aquaculture. Martínez Patiño, D.; Blanco Pérez, J.C. Nóvoa Vázquez, S.; Ojea Martínez, J.

Course: Biodiversity of marine bottoms. Sánchez Mata, A. Design and Management of Bivalve Mollusk Hatcheries. Centro Internacional de Altos Estudios Course: pathological alterations and the immune Agronómicos Mediterráneos (CIHEAM). Zaragoza. system in bivalve mollusks. Carballal Durán, M.J.; Villalba García, A. Economy of hatchery production. Guerra Díaz, A.

Course: Molusk culture techniques. A study of the A Case study: the design of hatcheries for several incidence of bonamiosis. species in different media. Lovatelli, A.; Helm, M.; Montes Pérez, J.; Guerra Díaz, A. Guerra Díaz, A.

Program of Biodiversity and Conservation of the European/Latinamerican Week. AL-INVEST III. Natural Environment. BIC GALICIA. Commercialization and Technological Transfer in Extractive Fishery and Course: Biodiversity of marine bottoms. Sánchez Aquaculture. Vigo (Pontevedra). Mata, A. Marine research and development of aquaculture in Environment and Natural Resource Program. Galicia. Guerra Díaz, A.

Course: Biodiversity of marine bottoms. Sánchez Management of non-industrial Fishery through Mata, A. handling plans. Seminar organized by the General Secretariat of relations with the European and Doctoral Prigram of the University of Vigo: Foreign Cooperation of the Xunta de Galicia and the Biology of Organisms and Ecosystems. Spanish Agency for International Cooperation. Antigua (Guatemala). Molares Vila, J. Course: Ecology of open-ocean beaches on the coats of Galicia. Sánchez Mata, A. Computer Science for the management of Shellfish Beds. Seminar organized by the CPAM. Molares Course: Structure and organization of benthic com- Vila, J.; Rodriguez Ruibal, L. munities. Sánchez Mata, A.

COURSES

Undergraduate courses given in the Marine Science Program. University of Vigo

Marine Biology. 1st Course. Sánchez Mata, A.

Communities on the Coastal Zones. 4th Course. Sánchez Mata, A.

Biological Oceanography Methods. 3rd Course. Sánchez Mata, A.

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3.4.4. PARTICIPATION ON EXAMINING BOARDS TO JUDGE DOCTORAL CANDIDATES

Darriba Couñago, S. Montes Pérez, J. Title: A study of spermatogenesis and the gonadal Title: Study of the toxicity of the abalone (Haliotis cycle in Mytilus galloprovincialis with specific refer- tuberculata) on the Galician coast with special ence to the presence of a gonadal neoplasia and its attention to its cytohistochemical location. possible association with xenobiotics. Doctoral Candidate: Bravo Portela, I. Doctoral Candidate: Alonso Martínez, A. University de Vigo University of Vigo Title: Effects the protozoan parasite Perkinsus olseni Fuentes González, J.M. (atlanticus) on the Manila clam Ruditapes philip- Title: Evaluation of the susceptibility to disease, pinarum in Korea. immunological capability and productive traits in Doctoral Candidate: D. Kyung-Il Park populations of the European flat oyster Ostrea University de Santiago edulis of different geographic origin. Doctoral Candidate: da Silva Scardua, P.M. Title: A study of Vibrio parahaemolyticus in Galicia. Aquaculture Institute. University of Santiago de Clinical, molecular and ecological aspects. Compostela Doctoral Candidate: Bárbaro Lozano, A. University de Santiago Title: Genetic Analysis of the marine mussel Mytilus galloprovincialis Lmk. By means of proteomic tech- Title: Characterization of the emerging pathogen in niques. marine aquaculture, Pseudomonas anguilliseptica. Doctoral Candidate: Mosquera Gómez, E. Doctoral Candidate: López Romalde, S. Departament of Genetics. University of Santiago de University de Santiago Compostela

Guerra Díaz, A. ; Martinez Patiño, D. Title: Mollecular markers and sexual dimorphism in the genera Mytilus and Ensis (Mollusca, Bivalvia). Doctoral Candidate: Mariñas Pardo, L. A. University of A Coruña

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3.4.5. EDITION OF SCIENTIFIC PUBLICATIONS

Blanco Pérez, J. C. (editor in chief ) Salgado, C.; Fernández, M.; Molares, J. Revista Galega dos Recursos Mariños. Vilagarcía de Arousa (on line)

Guerra Díaz, A. (member of the editorial board) VII Foro dos Recursos Mariños e da Acuicultura das Rías Galegas, 7. ISBN: 84-609-5211-8

Sánchez Mata, A. JFAE, Journal of Food, Agriculture and Environment Scientific International Society of Food, Agriculture and Environment (ISFAE)

Villalba García, A. (member of the editorial board) Revista de Biología Marina y Oceanografía. Facultad de Ciencias del Mar, University of Valparaíso, Chile

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3.4.6.REVIEW OF ARTICLES IN SCIENTIFIC PUBLICATIONS

Aquatic Conservation: Marine and Freshwater

Ecosystems. Sánchez Mata, A.

Aquaculture. Fuentes González, J.M.

Boletín del IEO. Darriba Couñago, S. (3)

Ciencias Marinas. Guerra Díaz, A. (2).

Ciencias Marinas. Montes Pérez, J.

Ciencias Marinas. Carballal Durán, M.J.

Capítulo de parasitología, de libro Biology and cul- ture of molluscs Nodipecten.

Journal of the Marine Biological Association of the UK. Sánchez Mata, A.

Journal of Applied Ichthyology. Sánchez Mata, A.

Journal of the Marine Biological Association of the UK (JMBA). Darriba Couñago, S.

Journal of Invertebrate Pathology. Villalba García, A.

Marine Biology. Villalba García, A.

OIE Manual of Diagnostic Tests for Aquatic Animals, 5th Edition. Villalba García, A

Scientia Marina. International Journal on Marine Sciences. Molares Vila, J.

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3.4.7. PATENTS

Patent Title: Antibodies of mice and their application in the spe- cific identification of larva D in the mussle Mytilus galloprovincialis.

Applicants: University ofVigo and Xunta de Galicia

Inventors: González Fernández, A. Lorenzo Abalde, S. Fuentes González, J. M.

OEPM Application Number: P200300629

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3.5. STAFF TRAINING

3.5.1. VISITS TO OTHER CENTERS

Cao Hermida, M.A.; Casas Liste, S.M. Comesaña Lestayo, P. Subject: Analysis of the immunological parameters Subject: An analysis of the immunological parame- of Ostrea edulis and Crassostrea gigas. ters of Ostrea edulis and Crassotrea gigas. Duration: 7 days Duration: 7 days Center: University Collage Cork (Ireland). Centro: University Collage Cork. Ireland

Cerviño Otero, A. Subject: Molecular techniques to diagnose bonamio- Subject: Trials carried out in the culture of the pullet sis. Analysis of the immunological parameters of the carpet shell clam. oyster Crassostrea ariakensis relative to bonamiosis. Duration: 4 days Duration: 1 month Center: Molllusk Hatchery in Tinamenor Centro: Virginia Institute of Marine Science. (Santander). Gloucester Point. Virginia. EEUU.

Sánchez Mata, A. Subject: Spanish Antarctic Survey 2004-2005. Sampling and lab processing of sediments and biots for the análisis of Antarctic intertidal benthos. Duration: 2 months Center: Spanish Antarctic Base Juan Carlos I, Livingston Island and Spanish Antarctic Base Gabriel de Castilla, Deception Island, Shetland Islands in the South Antarctic.

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3.5.2. COURSE ATTENDANCE

Design and Management of Bivalve Mollusc Curso para a Obtención do Diploma de Directivo da Hatcheries. CIHEAM. FAO. Zaragoza. Xunta de Galicia. Cerviño Otero, A. Molares Vila, J.

Procedemento Administrativo na Modalidade de Taller de Nuevas Tecnologías para el Análisis Teleformación. EGAP. Xunta de Galicia. Territorial: Puesta en marcha de un Project de siste- Cruzado Estévez, A.; Ventoso Padín, P. mas de información geográfica. San Sebastián. Carreira Vázquez, P.; Martínez Verde, G.; Rodal Gestión y Tratamiento de Residuos en la Mallo, M.; Rodríguez Ruibal, L. Acuicultura. 1º Seminario. Vigo (Pontevedra). Alonso Fdez. de Landa, J.L.; Darriba Couñago S.; Sistemas de Información Geográfica y Sistemas de Guerra Díaz, A.; Miranda Bamio, M.; Posicionamiento Global (GPS) en Nuestra Vida Cotidiana. San Sebastián. Xestor de Bases de Datos Relacionais. EGAP. Xunta Carreira Vázquez, P.; Martínez Verde, G.; Rodal de Galicia. Mallo, M.; Rodríguez Ruibal, L. Gregorio Chenlo, M.V. Uso de los Sistemas de Información Geográfica en Course de Iniciación a la Biología Marina. Pesca y Acuicultura. Zaragoza. L’Oceanografic (Ciudad de las Artes y las Ciencias). Parada Encisa, J.M. Valencia. Otero Otero, M. Curso de Introducción al Uso de las Bases de Datos de la Web of Knowledge. Fundación Española II Curso sobre Etología y Mantenimiento de Ciencia y Tecnología. Vigo. Mamíferos. Aquarium Finisterrae (Museos Cientí- Darriba Couñago, S. ficos Coruñeses). A Coruña. Otero Otero, M. V Curso de Ictiopatología Práctica para Piscicultores: situación sanitaria actual del cultivo de rodaballo y Recolectora de Algas Comerciais. Consellería de lenguado. CETMAR , IGAFA, SKRETTING. Vigo. Pesca e Asuntos Marítimos. Illa de Arousa. Pazos Sieira, G.; Soto Fraga, N. Otero Otero, M.; Búa Santamaría, I. VALBIOMAR, Course de presentación de alternativas Curso de Perfeccionamento na Lingua Galega. para el uso y aprovechamiento de residuos proce- Concello de O Grove. dentes de la pesca y acuicultura. CETMAR (Vigo). Otero Otero, M. Guerrero Valero, S.

Hidrolizados de Proteínas de Pescado y Ensilados. VALBIOMAR II. Hidrolizados proteicos de pescado y Proxecto Valbiomar. Centro Tecnolóxico do Mar ensilados: características y propiedades, preparación (CETMAR). y aplicaciones para la acuicultura. CETMAR (Vigo). Otero Otero, M. Guerrero Valero, S.

Avaliación de Contaminantes Biolóxicos. EGAP. Xunta de Galicia. Miranda Bamio, M.

Jornada sobre: Avances Científicos en la Investigación de Salmonella y Vibrio parahaemoly- ticus. CETMAR. Vigo (Pontevedra). Darriba Couñago, S.; Miranda Bamio, M.

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3.6. COLLABORATION AGREEMENTS

Collaboration agreement between CETMAR and the aimed at the management of specific resources CPAM to procide technical assistance and the devel- (SIGREMAR) to other shellfish resources in Galicia”. opment of the following projects and research actions. Mapping and evaluation of zones and Collaboration agreement with the University of specie sof interest in the action plan for the recovery Concepción (Chile) for Project FONDEF. of the Lombos do Ulla area; Culture and manage- ment of solenaceans. Collaboration agreement between the CPAM and the Organization of Mussel farmers of Galicia Framework Cooperation Agreement CPMA-ANFA- (OPMEGA) to carry our a study on the recruitment of CO. the mussel on collector ropes within the context of the research action “Evaluation of the larval abun- Framework Cooperation Agreement with the dance and recruitment of mussel spat Mytilus gallo- University of A Coruña. provincialis on the Galician coast”.

Framework Cooperation Agreement with the Collaboration agreement with the USC on the proj- University of Vigo. ect: “Techniques for the minimization, treatment and use of waste from aquaculture”. Subproject: Collaboration agreement between the CPAM and “Study on the evaluation of fish by-products from the Aquaculture Association “Illa do Santo” of Bueu fish farms. Biochemical and microbiological aspects.· (Pontevedra) to farm fishes in cages.

Collaboration agreement with the IEO to develop a research project.

Collaboration agreement between the CPAM and the CESGA to carry out the research project entitled “Extension of the Geographic Information System

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3.7. OUTSIDE VISITORS

CIMA in Vilanova de Arousa

Dr. Cesar Lodeiros Sabbatical Year Instituto Oceanográfico de Venezuela. University of Oriente. Venezuela

Veronique Sechet IFREMER Nantes ASP Toxicity in scallops.

Yanet Guerrero Rentería Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE). Ensanada. Baja California. Mexico. Training in the diagnosis of disease in mollusks

CIMA in Ribadeo

Adele Cromie Queen´s University Belfast Centre for Marine Resources and Mariculture 1 month

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3.8. PUBLICATIONS

3.8.1. ARTICLES IN JOURNALS LISTED IN THE Science Citation Index (SCI)

Abollo, E.; Pascual, S. (2005). Renal coccidiosis in the Li, A.; D’Amelio, S.; Paggi, L.; He, F.; Gasser, R.; Lun, European cormorant Phalacrocórax aristotelis aris- Z.; Abollo, E.; Turchetto, M.; Zhu, X. (2005). Genetic totelis from the Galician coast. Journal of Marine evidence for the existence of sibling species within Biological Association of the United Kingdom, 85: Contracaecum rudolphii (Hartwich, 1964) and the 1017-1019. validity of Contracaecum septentrionale (Kreis, 1955) (Nematoda: Anisakidae). Parasitology Carballal, M. J.; Díaz S.; Villalba, A. (2005). Research, 96: 361-366. Urosporidium sp. hyperparasite of the turbellarian Paravortex cardii in the cockle Cerastoderma edule. López, J.L.; Lorenzo-Abalde, S.; Fuentes, J. (2005). Journal Invertebrate Pathology, 90(2): 104-107. Proteomic approach to probe for larval proteins of the mussel Mytilus galloprovincialis. Marine Da Silva, P.M.; Fuentes, J.; Villalba, A. (2005). Biotechnology, 7 : 396-404. Growth, mortality and disease susceptibility of oys- ter Ostrea edulis families obtained from brood Lorenzo-Abalde, S.; González-Fernández, A.; de stocks of different geographical origins, through on Miguel-Villegas, E.; Fuentes, J. (2005). Two monoclo- growing in the Ría de Arousa (Galicia, NW Spain). nal antibodies for the recognition of Mytilus spp. Marine Biology, 145: 965-977. larvae: studies on cultured larvae and test on plank- ton samples. Aquaculture, 250: 736-747. Da Silva, P. M. ; Soudant, P.; Carballal, M. J.; Lambert, C. ; Villalba A. (2005). Flow cytometric Macho, G.; Molares, J.; Vázquez, E. (2005). Timing of DNA content analysis of neoplastic cells in larval release by three barnacles from the NW haemolymph of the cockle Cerastoderma edule. Iberian Peninsula. Marine Ecology Progress Series, Diseases of Aquatic Organisms, 67: 133-139. 298: 251-260.

Da Silva, P.M.; Villalba, A.; Sunila, I. (en prensa). Mc Donough, N.A. and Patiño, D.M. (2005). Branchial lesions associated with abundant apoptot- Developing stock enhancement techniques for two ic cells in oysters Ostrea edulis of Galicia (NW Spain). razor clams species en the European Atlantic Area. Diseases of Aquatic Organisms. Journal of Shellfhis Research. 24, (1): 329.

Darriba, S.; San Juan, F.; Guerra, A. (2005). Pascual, S.; Abollo, E. (2005). Whaleworms as a tag Gametogenesis cycle of Ensis siliqua (Linnaeus, 1758) to map heavy metal pollution zones. Trends in in the Ría de Corcubión, Northwestern Spain. Parasitology, 21: 204-206. Journal of Molluscan Studies, 71: 47-51 Pascual, S.; Abollo, E.; Azevedo, C. (en prensa). Host- Darriba, S.; San Juan, F.; Guerra, A. (2005). Energy parasite interaction of a muscular-infecting didymo- storage and utilization in relation to reproductive zoid in the musculature of the Atlantic mackerel cycle in the razor clam Ensis arcuatus (Jeffreys, Scomber scombrus L. ICES Journal of Marine Science. 1865). ICES Journal of Marine Science, 62: 886-896. Prado, S.; Romalde, J.L.; Montes, J.; Barja, J.L. (2005). Kroeck, M.A.; Montes, J. (2005). Occurrence of the Pathogenic bacteria isolated from disease outbreaks haemocyte parasite, Bonamia sp., in flat oyster, in sellfish hatcheries. First description of Vibrio nep- Ostrea puelchana d´Orbigny, farmed in San Antonio tunios as oyster pathogen. Diseases of Aquatic Bay (Argentina). Diseases of Aquatic Organisms, 62: Organisms, 67: 209-215. 231-235. Rodríguez, F.; Chauton, M.; Johnsen, G.; Andresen,

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K.; Zapata, M. (en prensa). Photoacclimation in phy- Villalba, A.; Casas, S.M.; López, C.; Carballal, M. J. toplankton: implications for biomass estimates, pig- (2005). Study of perkinsosis of the carpet shell clam ment functionality and chemotaxonomy. Marine Tapes decussatus in Galicia (NW Spain). II. Temporal Biology. pattern of diseases dynamics and association with clam mortality. Diseases of Aquatic Organisms, 65: 257-267.

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3.8.2. ARTICLES IN JOURNALS NOT LISTED IN THE SCI

Cerviño Eiroa, A; García, A; de Coo, A; Bao, M.; García Fernández, A; Cerviño Eiroa, A; De Coo Domínguez, M. Mejoras tecnológicas en el preen- Martín, A; Bao Iglesias, M.; Domínguez, M. (2005). gorde de moluscos bivalvos. Aplicaciones en el cultivo Tasa de crecimiento y mortalidad en el preengorde en batea en la Rías Bajas Gallegas. Galicia. España. de almeja fina, Ruditapes decussatus (LINNE, 1758), Actas del IX Congreso Nac. Acuicult.: 195-200. la almeja babosa, Venerupis pullastra (MONTAGU, 1803) y la almeja japonesa, Ruditapes philippinarum Conchas, R.F.; Montes, J. Bonamiosis control in (ADAMS & REEVE, 1850), en batea en las Rías Bajas. Galicia (NW Spain). Proceedings of European Actas del IX Congreso Nac. de Acuicult.: 211-213. Multicolloquium of Parasitology: 481-486. Guerrero, S.; Santos, I.; Outón, M.J. (2005). Mussel Conchas, R. F.; Alonso, B.; Montes, J. Patologías aso- and white shells by-products in Galicia, Spain. E.A.S. ciadas a los Solénidos: Ensis arcuatus, Jeffreys (nava- Special Publication, No. 35: 403-404. ja) y Ensis siliqua, Linneo (longueirón) en Galicia. Actas del IX Congreso Nac. Acuicult.: 253-256. Guerrero, S.; Santos, I.; Outón, M.J. (2005). Caracterización y cuantificación de los residuos pro- Conchas, R. F.; Alonso, B.; Montes, J. Oral vaccination cedentes de la acuicultura en Galicia. VIII Foro dos against vibriosis caused by Vibrio vulnificus serovar E Recursos Mariños e da Acuicultura nas Rias Galegas, in anguilli-culture. Actas del IX Congreso Nac. 7: 233-243. Acuicult.: 257-264. Linares, F.; Olmedo, M.; Peleteiro, J.B. (2005). Conchas, R.F.; Montes, J.; López, C.; Darriba, S.; Crecimiento y composición bioquímica de las larvas Iglesias, D. (2005). Bucephalidae trematode affect- de besugo (Pagelus bogaraveo), alimentadas con ing razor clam Ensis arcuatus from the coast of artemia enriquecida con tres microalgas diferentes. Galicia (NW Spain). Multidisciplinary for parasites, Actas del IX Congreso Nac. de Acuicult.: 353-357. vectors and parasitic diseases. Proceedings of European Multicolloquium of Parasitology: 505-509. López, C. ; Darriba, S.; Conchas, R.F. (2005). Presencia de Marteilia sp. en Solen marginatus en Galicia (NO Darriba, S.; Miranda, M.; da Costa, F.; Catoira, J.L. España). Acta Parasitológica Portuguesa. Revista da (2005). Estudios para la gestión de la explotación de Sociedade Portuguesa de Parasitologia, 12: 517-518. Solénidos. VIII Foro dos Recursos Mariños e da Acuicultura das Rías Galegas, 8: 215-220. Martínez, D.; Nóvoa, S.; Ojea, J.; Rodriguez-Moscoso, Darriba, S.; San Juan, F.; Guerra, A. (2005). E. (2005). Determinación de la talla de primera Metabolismo reproductivo de la navaja (Ensis arcua- maduración sexual de la coquina, Donax trunculus tus Jeffreys, 1865): Acumulación y movilización de (Linné, 1758) (Mollusca:Bivalvia), en un banco natu- lípidos de reserva durante el ciclo gametogénico. La ral de la Ría de O Barqueiro (NO de Galicia). Actas acuicultura como actividad económica en las zonas del IX Congreso Nac. de Acuicult.: 215-217. costeras. Actas del IX Congreso Nac. de Acuicult.: 201-204. Olmedo, M.; Peleteiro, J.B.; Cal, R.; Linares, F. Crecimiento de juveniles de lenguado (Solea senega- De Coo, A.; Cerviño-Eiroa, A.; García Fernández, A.; lensis, Kaup, 1858) en Galicia. Actas del IX Congreso Bao Iglesias, M.; Domínguez, M. (2005). Tasas de Nac. de Acuicult.: 303-304. crecimiento, formación de colas y mortalidad en el preengorde de la ostra plana (Ostrea edulis, Linné Santos, I.; Guerrero, S.; Outón, M.J. Turbot Farms 1758) en bateas en la Rías Bajas de Galicia. Su rela- Waste in Galicia, Spain. E.A.S. Special Publication No. ción con las densidades de cultivo y las operaciones 35: 235-236. de manejo. Actas del IX Congreso Nac. de Acuicult.: 195-200.

134 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005 SCIENTIFIC ACTIVITY

Chereguini, O.; García de la Banda, I.; Diez, J.; Lobo, García de la Banda, I.; Chereguini, O.; Lobo, C.; Diez, C.; Linares, F. Effect of different regimes on senegal J.; Linares, F.; Arce, F. Ongrowing of senegal sole sole Solea senegalensis (Kaup 1858) larval culture in Solea senegalensis (Kaup 1858) fries using two Cantabria (Northern Spain). World Aquaculture. Bali different commercial diets in Cantabria (Northern (Indonesia) Libro de actas. Spain). World Aquaculture. Bali (Indonesia) Libro de actas.

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3.8.3. CONTRIBUTIONS TO BOOKS

Blanco, J.; Moroño, A.; Fernández, M.L. Toxic Iniesta, R.; Blanco, J. Bioacumulación de Episodes in Shellfish, Produced by Lipophilic Hidrocarburos y Metales Asociados a Vertidos Phycotoxins: An Overview. Revista Galega dos Accidentales en Especies de Interés Comercial de Recursos Mariños (Monog.): 1-70 pp. (on line Galicia. Revista Galega dos Recursos Mariños http://www.recmar-galicia.org). (Monog.) 2-200 pp. (on line http://www.recmar-galicia.org). Garrido, J.L.; Zapata, M. Chlorophyll Analysis by New HPLC Methods. Chlorophylls and Bacteriochlo- Zapata, M.; Garrido, J.L.; Jeffrey, S.W. Chlorophylls rophylls: Biochemistry, Biophysics, Functionsand and Bacteriochlorophylls: Biochemistry, Biophysics, Applications, Grimm, B.; Porra, R.J.; Rüdiger, W.; Functions and Applications. Grimm, B.; Porra, R.J.; Scheer, H. (eds). Vol 25 of Advances in Rüdiger, W.; Scheer, H. (eds). Vol 25 of Advances in Photosynthesis and Respiration. Series Editor: Photosynthesis and Respiration. Series Editor: Govindjee, Springer, Dordrecht. ISBN: 1-4020-4515-8. Govindjee, Springer, Dordrecht. ISBN: 1-4020-4515-8.

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4. SCIENTIFIC MEETINGS AND TRAINING ORGANIZED BY THE CIMA

SCIENTIFIC MEETINGS AND TRAINING ORGANIZED BY THE CIMA

4.1. SEMINARS

Title: Protozoo parasitic Perkinsus marinus: new Title: The marine investigation and the technologi- solutions for an old problem?. cal transference to the companies of acuicultura in Speaker: Dr. José A. F. Robledo. University of Chile. Maryland Biotechnology (USA) Speaker: Dr. Irene Lépez García. Dpto. de Date: 19.05.05 Oceanografía. University de Concepción. Chile Date: 11.09.05

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5. ANNEX.

ANNEX

ABBREVIATIONS USED

AZTI Instituto Tecnológico Pesquero y Alimentario (Technological Institute of Fisheries and Food) CCGP Comité Científico Galego da Pesca (Galician Scientific Committee on Fisheries) CCMM Centro de Control de Calidade do Medio Mariño (Marine Monitoring Center) CESGA Centro de Supercomputación de Galicia (Adcanced computation center) CEP Centro de Experimentación Pesquera (Center for Fisheries Experiments) CETMAR Centro Tecnológico del Mar (Center for the Technology of the Sea) CIEHAM Centro Internacional de Altos Estudios Agronómicos del Mediterráneo de Zaragoza (International Center for Advanced Agronomical Studies of the Mediterranean of Zaragoza) CICEM Centro de Investigación para el Cultivo de las Especies Marinas (Research Center for the Culture of Marine Species) CIMA Centro de Investigacións Mariñas (Marine Research Center) CO Centro Oceanografico (Oceanographic Center) CPAM Consellería de Pesca e Asuntos Marítimos (Local Ministry for Fishery and Maritime Affairs) CSIC Consejo Superior de Investigaciones Marítimas (High Council for Maritime Research) DXIDP Dirección Xeral de Innovación e Desenvolvemento Pesqueiro (Directorate General of Innovation and Fisheries Development) DXRM Dirección Xeral de Recursos Mariños (Directorate General of Marine Resources) ICCM Instituto Canario de Ciencias Marinas (Canary Islands Institute for Marine Sciences) (Aquaculture Station) IEO Instituto Español de Oceanografía (Spanish Oceanography Institute) IGAFA Instituto Galego de Formación en Acuicultura (Galician Institute for Aquaculture Training) INTERREG Iniciativa Comunitaria de Cooperación Transfronteriza (Community Initiative for Transfrontier Cooperation) INTEGMAR Instituto Tecnológica del Mar (Technological Institute of the Sea) IPIMAR Instituto de Investição das Pescas e do Mar (Portugal) IRTA Institut de Recerca i Tecnología Agroalimentáries.

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JACUMAR Junta Asesora de Cultivos Marinos (Advisory Board for Marine Culture) (Harvesting Permit) SIGREMAR Sistema de Información Geográfico Orientado a la Gestión de Recursos Marinos. (Geographic Information System Oriented towards the Management of Marine Resources) UIMP Universidad Internacional Menéndez y Pelayo (International University of Menéndez y Pelayo) ULPGC Universidad de Palma de Gran Canaria (University of Palma of Gran Canaria) USC Universidad de Santiago de Compostela (University of Santiago de Compostela) UC Universidad de A Coruña (University of A Coruña) UO Universidad de Oviedo (University of Oviedo) UV Universidad de Vigo (University of Vigo)

146 CONSELLERÍA DE PESCA E ASUNTOS MARÍTIMOS CIMA Annual Report 2005