National Reference Laboratories for Mollusc Diseases
(Infection with Bonamia exitiosa, Bonamia ostreae, Marteilia refringens, Microcytos mackini, Perkinsus marinus)
Institute of Infectology
The national reference laboratory for mollusc diseases (NRL-Mu) is located at the Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, and is responsible for coordinating and ensuring the diagnosis of mollusc diseases, mainly parasitic diseases, based on the German regulation on fish diseases and the EU legislation. The EU Directive on Aquaculture lists Bonamiosis, caused by Bonamia ostrea and its variants, and Marteiliosis, caused by Marteilia refringens and its variants. Based on the recommendations of the Office International des Epizooties (OIE) another Bonamiosis, caused by Bonamia exitiosa, Microcytosis (Microcytos mackani) and Perkinsosis (Perkinsus marinus) must furthermore be diagnosed and differentiated. The NRL-Mu consists of the following NRLs: NRL for Bonamiosis, NRL for Marteiliosis, NRL for exotic parasites of molluscs (B. exitiosa, M. mackani, P. marinus). The NRL also investigates other mollusc diseases such as abalone diseases.
The diseases are notifiable in case of abnormal mortality in association with the detection of the pathogens.
In addition to parasitic diseases, irido-, herpes- and birnaviruses can be detected in molluscs.
Main research topics and projects
- Development of molecular diagnostic methods for detection of parasites and viruses of molluscs
- KHV-I carrier fish (Anodonta sp.)
Overview of services
- Direct contact for federal and state authorities with regard to the diagnosis of mollusc diseases
- Pathogen detection in sent-in samples
- Pathogen detection in case of abnormal mortality in so far disease-free areas or fish farms or confirmation of the results of regional laboratories
- Application and optimization of the standards, diagnostic procedures, and diagnostic reagents
- Testing, standardization and improvement of test procedures for detection of mollusc diseases
- Coordination of the standards and diagnostic procedures used by the diagnostic authorities for diagnosis of the respective diseases
- Organization of training measures for laboratory experts of the veterinary authorities
- Organization of comparative national ring tests
- Participation in comparative ring tests for EU Member States organized by the Community reference laboratory
- Annual report to the Community reference laboratory on epidemiological and laboratory data on the diagnosis and control of mollusc diseases
- Participation in the annual meeting of the representatives of the NRL-M of the EU Member States
Reference material
- Histological preparations of infected molluscs
- DNA and RNA preparations of pathogens (parasites and viruses)
Overview of methods
The standard procedures used by the NRL-M for detection of parasites of molluscs are:
- Formalin-fixed organs/molluscs
- Paraffin-embedding
- Histological sections in HE staining
- Smear preparations in HE and hemacolor staining
- PCR detection of parasites of molluscs
- Detection by RT-PCR and quantitative real-time PCR
- In-situ hybridization for detection of mollusc parasites and viruses
Other methods used by the laboratory
- Chromatography (e.g. gel filtration, affinity chromatography)
- Agar gel precipitation test (AGPT)
- Electrophoresis (e.g. immunoelectrophoresis, PAGE, IF, 2-D electrophoresis)
- Cell culture techniques for fish, mollusc and mammalian viruses
- Western and Dot Blot
- Cloning and sequencing
- Transfection
- Virus purification and concentration
- ELISA (antigen and antibody)
- PCR, RT-PCR, real-time PCR, In-situ hybridization
Relevant legislation
Council Directive 2006/88/EC on animal health requirements for aquaculture animals and products thereof, and on the prevention and control of certain diseases in aquatic animals (Official Journal of the EU No. L328 p. 14)
Commission Decision 2001/183/EC dd. 22 February 2001 laying down the sampling plans and diagnostic methods for the detection and confirmation of certain fish diseases and repealing Decision 927532/EEC. (Official Journal of the EU No. L 56 p. 65)
Regulation on fish diseases and regulation on the modification of the regulation on notifiable animal diseases dd. 22 November 2008 (Federal Gazette I p. 2315)
Regulation on notifiable animal diseases dd. 3 November 2004, last modified by the Second Regulation on the modification of animal disease regulations dd 18 December 2009.
in their latest version