Project no. SSPE-CT-2003-502329 PANDA Permanent network to strengthen expertise on infectious diseases of aquaculture species and scientific advice to EU policy Coordination Action, Scientific support to policies WP4: Report on the current best methods for rapid and accurate detection of the main disease hazards in aquaculture, requirements for improvement, their eventual standardisation and validation, and how to achieve harmonised implementation throughout Europe of the best diagnostic methods Olga Haenen*, Inger Dalsgaard, Jean-Robert Bonami, Jean-Pierre Joly, Niels Olesen, Britt Bang Jensen, Ellen Ariel, Laurence Miossec and Isabelle Arzul Work package leader & corresponding author: Dr Olga Haenen, CIDC-Lelystad, NL ([email protected]) PANDA co-ordinator: Dr Barry Hill, CEFAS, UK; www.europanda.net © PANDA, 2007 Cover image: Koi with Koi Herpes Virus Disease: enophthalmia and gill necrosis (M.Engelsma acknowl.) Contents Executive summary 5 Section 1 Introduction 7 1.1 Description of work 7 1.2 Deliverables 8 1.3 Milestones and expected results 9 1.4 Structure of the report and how to use it 9 1.5 General remarks and links with other WPs of PANDA 9 Section 2 Materials and methods 10 2.1 Task force 10 2.2 Network 10 2.3 Workshops and dissemination 10 2.4 Analysis of data 10 2.5 Why harmonization throughout Europe background and aim 11 2.6. CRL functions 11 Section 3 Results 12 3.1 Task force 12 3.2 Network 12 3.3 Workshops and dissemination 12 3.4 Analysis of data 14 Diseases/pathogens of fish 14 3.4.1 Epizootic haematopoietic necrosis virus (EHN) 14 3.4.2 Red sea bream iridovirus 16 3.4.3 Infectious Salmon Anaemia Virus 19 3.4.4 Koi Herpes Virus 22 3.4.5 Streptococcus agalactiae 27 3.4.6 Streptococcus iniae 29 3.4.7 Lactococcus garvieae 32 3.4.8 Trypanoplasma salmositica 34 3.4.9 Ceratomyxa shasta 38 3.4.10 Neoparamoeba perurans 40 3.4.11 Parvicapsula pseudobranchicola 43 3.4.12 Gyrodactylus salaris (Gyrodactylosis) 44 3.4.13 Aphanomyces invadans (Epizootic Ulcerative Syndrome) 48 3 Diseases/pathogens of molluscs 48 3.4.14 Introduction on mollusc diseases/pathogens 52 3.4.15 Nocardia spp. (Pacific oyster nocardiosis – Nocardia crassostreae ) 53 3.4.16 Candidatus Xenohaliotis californiensis (withering syndrome) 54 3.4.17 Perkinsus olseni /atlanticus 55 3.4.18 Perkinsus marinus 57 3.4.19 Marteilioides spp. ( M. chungmuensis : Marteilioidosis) 58 3.4.20 References for all molluscs diseases in this report 60 Diseases/pathogens of crustaceans 62 3.4.21 Introduction to crustacean diseases/pathogens 62 3.4.22 Yellowhead disease (YHD) 62 3.4.23 White spot disease (WSD or WSSD), White spot syndrome (WSS) 64 3.4.24 Taura Syndrome (TS) 66 3.4.25 Infectious hypodermal and haematopoietic necrosis (IHHN) 68 3.4.26 Coxiella cheraxi 70 3.4.27 References of crustacean pathogens/diseases: 71 Diseases/pathogens of amphibians 73 3.4.28 Amphibian ranavirus 73 3.4.29 Batrachochytrium dendrobatidis (amphibian chytridiomycosis) 75 Section 4 General discussion 79 4.1 Evaluation of available tests and recommendations for improvement 79 4.2 Priority list for improvement of tests/testing per pathogen/disease, 86 and skills 4.3 Working towards standardization and validation 87 4.4 General remarks and links with other WPs 87 4.5 Recommendations to achieve harmonized implementation 88 Section 5 General conclusions and recommendations 91 Section 6 Acknowledgements 92 Section 7 Annexes 93 7.1 Task force 93 7.2 List of consulted experts per pathogen 94 7.3 Table on methods described in literature for detection of ISAV and 95 diagnosis of ISA 7.4 Current available detection and diagnostic methods for some 98 non WP2 listed diseases/pathogens of molluscs and crustaceans 7.4.1 Herpesvirus (oyster herpes-like virus disease, OsHV1) 98 7.4.2 Bonamia ostreae 100 7.4.3 Marteilia refringens 102 7.4.4 Gaffkemia ( Aerococcus viridans ) 104 7.4.5 Crayfish plague ( Aphanomyces astaci ) 104 7.5 Table on the evaluation of available methods for non WP2 listed mollusc and 106 crustacean diseases/pathogens 7.6 List of illustrations and authors 107 7.7 The PANDA consortium 108 4 Executive summary Within the PANDA project, the objectives of Work tests for important and current EC listed viruses (Viral Package 4 (WP4) are to identify the optimal diagnostic Haemorrhagic Septicaemia Virus (VHSV), Infectious methods currently available for the most serious Haematopoietic Necrosis Virus (IHNV) and Spring diseases, which were identified by risk analysis (WP2), Viraemia of Carp Virus (SVCV)). Extension of the and to provide recommendations for their standardis - training and ring tests with the fish pathogens EHNV, ation and harmonisation procedures throughout KHV and EUS is advized, apart from with Infectious Europe and for any needs to improve their accuracy, Salmon Anaemia Virus (ISAV). For the 3 mentioned fish rapidity and applicability. bacteria, fast and accurate additional tests are needed for confirmation. For the 4 fish parasites, expertise The work package was lead by participant 4, Olga lacks in Europe, to screen for these parasites, and type Haenen, who appointed a task force to do the work them. However, as these parasites are not listed yet together. The WP4 task force made tables with current by EC or OIE, they have a lower priority. available diagnostic methods, with literature refer - ences, and if the test were well established or The mollusc diseases and pathogens are well validated. These tables were put at the panda website covered in expertise and training via the CRL for to get input from PANDA network members. Mollusc Diseases, which organizes workshops on Additionally, the draft tables were sent to leading endemic and exotic important diseases and pathogens experts per pathogen, which were asked for for NRL’s. Furthermore, they already take the exotic comments. From this latter action many input pathogens into account in their ring test. resulted. During the plenary PANDA workshops in For crustacean diagnosis , appointment of a CRL by Lelystad (April 2006) and Weymouth (March 2007), the the EC is necessary, and training on clinics and WP4 results were discussed, and plans for making the diagnosis of Yellowhead disease, White Spot Disease, reports were made. During the whole project time, and Taura syndrome is recommended. The task force many lectures were presented to international furthermore recommended to acquire expertise and audiences, and many experts joined the PANDA testing for the non-WP2 listed Crayfish plague by network. Aphanomyces astaci , as this disease is a threat for From the tables and the discussions, this report was crustaceans all over Europe. made. It was concluded, there are many well estab - The amphibian diseases /pathogens RANA virus and lished tests for diagnosis of disease and detection of Batrachochytrium dendrobatidis , a fungus are new to hazardous pathogens of aquaculture species, like most laboratories. Appointment of a CRL by the EC is defined in the WP2 list. However, many of the necessary, after which certain laboratories should get diseases or pathogens are not known yet by most expertise and skills in testing via training. Many of the laboratories in the EC. internationally available tests are non validated, but for For fish diseases , especially for the recently EC-listed reason of daily use at laboratories well established. aquaculture diseases, acquisition of expertise into the However, these tests need validation and ring testing, EC, and training in screening and diagnostic after they have been implemented into European techniques on the viruses Epizootic Haematopoietic laboratories. Necrosis (EHN), Koi Herpes Virus Disease (KHVD), With the new lists of diseases of the EC and WP2 of Epizootic Ulcerative Syndrome (EUS) was recom - PANDA, the tasks to achieve harmonised implemen - mended. The Community Reference Laboratory on tation throughout Europe of the best methods are Fish Diseases sofar organizes workshops and ring extended for several responsible bodies: The 5 European Commission, Community Reference Laboratory, and the National Reference Laboratory with their government will have to put much effort and money, using the PANDA network and world wide experts, to get the expertise into Europe and to the CRL’s, NRL’s and regional labs. Priorities have to be made in the whole process, and therefore ad hoc expert groups need to be appointed first. In this way, the PANDA network can be further used. 6 section 1 Introduction The aquaculture industry is growing, both at European Through this network discussion, information was and at world level. More and more globalisation takes assimilated and collated on: the current status of place. This includes transport of live aquaculture available diagnostic methods for the diseases animals all over the world. These transports carry the identified in WP2 (Table 1.1.a), new developments in risk of transporting aquaculture disease as well, and methods for disease diagnosis and surveillance, with introducing these diseases into a new region. The EC an assessment of their specificity, sensitivity and has good legislation to be able to trade relatively safe, speed and their potential applicability to diseases by the current Aquaculture Directive 91/67/EC, and by identified in WP2, needs for validation and standardis - implementing the EC Directive 2006/88/EC. ation of diagnostic methods for the serious aquatic animal diseases, needs for strengthening knowledge Within the PANDA project, work package 2 has and technical skills to achieve harmonised application identified the most serious aquaculture diseases and within the EU for the current best diagnostic methods hazards, which threaten European aquaculture: exotic, identified. emerging and re-emerging disease hazards of potential risk to Europe, including an assessment of Network & Workshops: Workshops of the task force their potential impact on aquaculture and aquatic and other experts selected from the network were wildlife in the EU.