World Bank Business Plan Authors: Èož-Rakovac, Strunjak-Peroviæ, Hacmanjek, Topiæ Popoviæ
Rudjer Boskovic Institute
Project of the Ichthyopathology Group
Rudjer Boskovic Institute
Center for Marine and Environmental Research
Bijenicka 54, Zagreb, Croatia
Tel. 098 417607, 091 5602887,
Fax. 01 4854990
Ichthyopathology Group:
Dr. Rozelindra Èož-Rakovac, PhD, DVM
Dr. Mato Hacmanjek, PhD, DVM
Dr. Ivanèica Strunjak-Peroviæ, PhD, DVM
Dr. Natalija Topiæ Popoviæ, MS, DVM
1. Introduction and Background
We are hereby submitting proposal for support of ongoing and future scientific and technical programs that are of great economic importance for Croatia. Based on the knowledge and experience of our group we propose to establish an Authorized Diagnostic Center for Aquatic Animal Diseases within the Rudjer Boskovic Institute, for which we will implement legislative measures in accordance with the European Union Standards. The Center will:
1.Carry out both fundamental and applied research for fish health control and ecosystem protection
2.Offer diagnostic services to the Croatian fish farming industry
3.Offer educational services to the Croatian fish farming industry.
The fish farming industry is one of the fastest growing food producing sectors of the world. This industry has recently started to expand in Croatia and has the potential to contribute significantly to the economy. Major production constraints faced by the fish farming industry are outbreaks of disease and the general health of the farmed populations. These represent both ecological and economical issues.
Currently, Croatia is a pristine and virtually intact oasis of clean water resources. Constant monitoring is required to maintain this state and also to ensure the efficient growth and viability of the emerging fish farming industry.
The Center would reduce the direct cost of disease impacts of aquaculture and commercial fisheries, protect the competitiveness of Croatia’s export products and improve public confidence in the health of aquatic animals and security of food supply.
According to the EU legislative criteria will be set for Quality Assurance/Quality Control mechanisms for disease diagnostic laboratories.
2. Mission
As detailed below, the mission of the Center proposed herein is manifold
2.1. Fundamental & Applied Research for Fish Health Control and Protection of Ecosystems
In order to be able to control the health of farmed fish, thereby increasing the efficiency of production and to also preserve and maintain the supporting aquatic ecosystem, it is critical to understand fundamental aspects of physiology. Key physiological factors that must be researched are detailed in Sections 2.1.1 – 2.1.4 below.
2.1.1. Blood Biochemistry
Fish can experience biological and abiotic stresses from both natural and from anthropogenic sources. Such stresses can directly compromise fish health and if severe enough can cause disease or death. Biochemical and molecular parameters are the first indicators of homeostatic imbalance. Therefore, we propose the routine examination of farmed fish blood (plasma chemistry) to rapidly diagnose and treat changes in fish health status. To efficiently perform such screening standard plasma chemistry values must be established for farmed fish species.
2.1.2. Drug Interactions and Design
Improved diagnostic methods to detect fish diseases are required along with better ways to control and treat such diseases. The indiscriminate use of drugs must be avoided since there is a risk that drug resistant strains of the pathogens will develop. For this reason the use of antibiotics should be limited and other control methods used in their place. Veterinarians in Croatia have authority for the off-label use of approved animal drugs. If drug products are intended for and are used to treat a food-producing animal (e.g. fish), then residues in the food products must also be established and demonstrated to be safe for human consumption. Therefore, we intend to conduct a series of well-controlled studies to compare the depletion kinetics of drugs from the different tissues of series of farmed fish species. Such studies will include dose ranging in order to determine the optimal dosage for treating specific species of diseased farmed fish. Such studies would contribute greatly to guidance on the use and routes of administration of «aquaculture» drugs.
2.1.3. Determination of the Hepatic Xenobiotic Metabolizing Enzyme Systems in Fish
Since the cytochrome P450 isoenzymes (e.g. CYP1A & CYP3A) metabolize xenobiotics (pollutants and drugs) in fish as well as in mammals, they can act as a control point influencing drug plasma level, half-life, elimination or final effect of drugs. We propose to evaluate the effect of some antimicrobials used for fish treatment on CYP1A and CYP3A induction, kinetics and protein content in the tissues of fish farmed in Croatia.
2.1.4. Macroscopic and Microscopic Anatomy of Aquatic Organisms
As a first step in the development of rapid diagnostic methods for fish diseases, we propose to conduct an extensive study to define the normal physiological and pathological features of tissues from a variety of farmed fish in Croatia. Macroscopic and microscopic analysis of histological sections from different organs in different life stages of the various fish species would be evaluated. In combination with other diagnostic methods such as plasma chemistry (Section 2.1.1 above), we would establish the ranges of normal features and set the standards for rapid definition of pathological changes in locally farmed fish. This would be very important for future diagnosis and treatment of farmed fish populations.
2.2. Technological Development
2.2.1. Authorized Diagnostic Laboratory for Aquatic Organisms (reared and wild) Diseases
Based on the knowledge and experience of our group we propose to establish an Authorized Diagnostic Center for Fish Diseases within the Rudjer Boskovic Institute. The Center will offer diagnostic and educational services to the fish farming industry in Croatia and the region. Together with colleagues from the Croatian Veterinary Institute and Veterinary Faculty we propose to participate in the implementation of legislative measures in accordance with the European Union standards for diagnostics and treatment of fish diseases (International Aquatic Animal Health Code of the Office International des Epizooties – OIE 2000).
A critical element will be the evaluation of the epizootiological situation in Croatia. This will require systematic investigations of given populations of aquatic animals take place identifying surveillance zones for disease control purposes.
2.2.2. Authorized Diagnostic Laboratory for Pet Aquarium Fishes
Many people keep pet fish aquariums in their homes and there are many pet shops that import fish and aquatic animals. The poor management of tanks in homes and pet shops and the potential import of pathogens with fish and aquatic animals brought into Croatia from other parts of the world represents a potential threat to the fish farming industry and the aquatic ecosystem. Therefore, we propose to carry out a research study on parasitic diseases that cause the majority of death to fish in pet aquaria. For the purpose of the study, and with the help of media, local pet-fish owners would be encouraged to bring their dead and moribund fish to our Center for examination. After data collection and analyses, measures to treat, mitigate and or prevent such diseases would be recommended and made accessible to veterinarians, pet-shop owners, inspectors and fish owners.
2.2.3. Application of Available Human Rapid Identification Kits for Ichthyopathology
We intend to evaluate the effectiveness of the BBL E/NF (Becton Dickinson, USA) rapid identification system for implementation in screening of possible bacterial fish pathogens. To date, the BBL E/NF system has not been used aquatic organism microbiology, unlike some other rapid tests, like API (Biomerierux, France), Mono-kits (Bionor, Norway), GN Microplates (Biolog, USA), which all have shortcomings for application to fish bacteria identification.
2.2.4. Organization and Establishment of Fish Blood Bank
There is a need to establish a bank that would hold fish blood obtained from various fish species in different age groups. The blood would be catalogued and available for research and educational purposes for scientists throughout the world. Standard values would be established and blood would also be a resource for molecular research and diagnostics.
2.2.5. Assessment of ecological risks; design, development and implementation of monitoring systems for biological parameters
2.3. Education
Since most of the fish disease problems can be avoided with proper management, we intend to organize a series of workshops for fish farmers, veterinarians, inspectors and pet fish owners. Such workshops will teach:
1.Procedures to avoid diseases
2.How to respond to disease problems
3.Correct ways of shipping fish to a disease diagnostic laboratory
4.Treatment of diseased fish
5.Education on threatening and also novel fish diseases and conditions.
Histological images will be presented as an atlas on a web site to be available for comparison to other scientists and veterinarians in Croatia and world-wide.
One of the senior staff holds the position of graduate teacher at the Faculty of Agriculture in Ichthyohaematology science. Being a group of young, educated and experienced professionals and scientists in the fields of oceanology, ichthyology and veterinary fish medicine, we as a Group would like to respond to the needs and requests for higher education of colleagues who aspire to MS and PhD degrees by being their mentors and advisers.
Our group intends to form consultancy services for health/disease control. Most often, the nature of fish health issues is one of law and/or regulation. Such consultancy services will be provided within the framework of existing regulations and laws.
2.4. RBI Services
The existing Group already co-operates with a number of industrial Fish Farming clients. The intent of the proposed Center is to expand this client base offering services, including the training of management and staff at fish farms. This educational training will comprise on-site lectures and problem-solving that should lead to improvement of productivity by prevention and treatment of diseases. The Center will also provide direct technical and scientific assistance as required.
In addition, the existing Group has extensive collaborations with international experts and consultants (Cornell University, Ithaca, New York; Ashni Naturaceuticals, Ithaca, New York; National Institute of Biology, Ljubljana, Slovenia). It is conceivable that once established, the proposed Center will be able to extend its collaborations and provide services on an International basis.
However, to implement the proposed Center initial funding from the Croatian Ministry of Science and Technology and the World Bank is required.
2.5. Future
By the year 2010 it is envisioned that the proposed Center will be:
1.An internationally recognized Center of Excellence for the diagnosis, prevention and treatment of diseases of fish, shellfish and marine invertebrates.
2.A developer of novel diagnostic methodologies that are in concordance with the European Union (EU) regulations and standards.
3.An organization that helps to set policy within the EU
4.An internationally recognized Center of Excellence for fundamental and applied research of aquatic animal diseases.
5.Providing expert contract research services to biotechnology and pharmaceutical companies involved in aquatic veterinarian drug development programs
6.A key educational network site providing workshops, seminars & higher education qualifications.
7.Cooperating on a world-wide basis with other diagnostic centers and research groups.
8.A key member and participant in the EAFP (European Association of Fish Pathologists) and other international organizations.
3. Current situation
3.1. SWOT Analysis (Strengths, Weaknesses, Opportunities, Threats)
Strengths
7, 10, 15 and 20 years respective experience of the proposing professional and scientific experience
Existing collaboration with international experts
Established high reputation in the international scientific community and with industrial customers
Demonstrated strengths at the leading edge of the research and development of biochemical and molecular methods in aquatic animal science (see publication list in Section 5.2)
Commercial applicability to the drug development industry, the food technology industry and to the protection of human and animal health
Enthusiastic, motivated and professional team
Weaknesses
Need for initial financial input for equipping the Laboratory and attending professional meetings and symposia
Additional technical staff are required
Opportunities
Potential to become a world recognized center of Diagnostic & Research Excellence for fish, shellfish and marine invertebrates
Potential to develop and implement new commercializable methodologies in concordance with the EU and other international regulations and standards
Implementation of molecular science in diagnostics, prevention and treatment of aquatic animal diseases
Potential to develop collaborations and co-operate with relevant European and world-wide diagnostic and research centers
Improve the efficiency, economics and productivity of the fish farming industry in Croatia
Commercial applications providing expert contract research and development services to industrial clients
Potential to generate intellectual property (IP) that may lead to spin-off companies from the Center or that could be licensed to industrial companies
Threats
Unstable economic situation in the country
Need for competition with developed and leading countries with higher budget for R&D
3.2. PEST Analysis (Political, Economic, Social and Technological Factors)
Political Factors
Our existence as a research group is based on the current policy of the Ministry of Science and Technology
EconomicFactors
Unstable economic situation in the country
Croatia is a small, relatively undeveloped country and has minor resources for independent grant providers that could support activities of the Group
Limitation of the creative thinking because of the funneling of research interests by the financiers
Social Factors
Uncertain job positions
Technological Factors
Related to economical factors
4. Market Plan
Establish a Center of Excellence for the standardization and implementation of diagnostic techniques for our designated activities
Provide expert diagnostic and treatment services to the Croatian Fish farming (freshwater, marine, aquaria) and fishing industries (fish as human food)
Provide ecological risks and ecological impact assessments for different customers
Provide educational services to the above clients and to official veterinary inspectors
Organize new educational opportunities for professionals and graduate studies, including workshops, seminars and post-graduate studies
Provide expert contract drug research and development services to clients from the pharmaceutical and biotechnology industries
Generate IP that can be licensed to industrial companies or form the basis of spin-out companies, thereby generating revenues for the Center.
5. Human Resources
All staff graduated from the University of Zagreb (Veterinary Faculty). All received their MS degrees at the University of Zagreb (Faculty of Science) in the field of Oceanology. The first three members defended their PhD theses in the field of Biochemistry, Virology and Parasitology at the University of Zagreb (Veterinary Faculty). One member is a Fulbright scholar with Cornell University research experience.
All participated in scientific projects:
1.Genetics, nutrition and pathology of aquatic animals
2.Aquatic organisms as bioindicators of water quality in which they live
Project “Systematic research of the Adriatic Sea as a base for sustainable development of the Republic of Croatia (shortly “Project Adriatic”) and have extensive experience in laboratory and field diagnostic work. All have membership of the Croatian Microbiological Society and the Croatian Veterinary Association
5.1. Existing Collaborations with Experts from Croatian Institutions:
Veterinary Faculty, University of Zagreb
Faculty of Agriculture, University of Zagreb
Faculty of Food Technology and Biotechnology, University of Zagreb
Croatian Veterinary Institute, Zagreb
Public Health Institute of Republic of Croatia, Zagreb
Oikon doo, environment protection company of Zagreb
5.2. Recent Research Papers
Èož-Rakovac, Rozelindra; Teskeredžiæ, Emin; Hacmanjek, Mato; Tomec, Marija; Topiæ- Popoviæ, Natalija; Strunjak-Peroviæ, Ivanèica: Seawater challenge test and body composition of Coho salmon (Onchorynchus kisutch). Veterinarni Medicina. 44 (1999), 12; 365-371.
Topiæ Popoviæ, Natalija; Strunjak-Peroviæ, Ivanèica; Fonns, A.; Vilsgaard-Espersen, T.; Teskeredžiæ, Emin: Pseudorhadinorhynchus salmothymi isolation from brown trout in Krka river (Croatia). Periodicum Biologorum. (1999), 3; 273-275.
Èož-Rakovac, Rozelindra; Topiæ Popoviæ, Natalija; Strunjak-Peroviæ, Ivanèica; Krèa, Sanja; Grubaèeviæ, Nedjeljka: Aeromonas hydrophila outburst on carp (Cyprinus carpio, L.) post transfer to aquarium. Periodicum Biologorum. 102 (2000), 3; 293-296.
Èož-Rakovac, Rozelindra; Teskeredžiæ, Emin: Biochemical changes in coho salmon plasma following sea water adaptation. Periodicum Biologorum. 1 (2000), 3; 297-301.
Topiæ Popoviæ, Natalija; Teskeredžiæ, Emin; Strunjak-Peroviæ, Ivanèica; Èož-Rakovac, Rozelindra: Aeromonas hydrophila isolated from wild freshwater fish in Croatia. Veterinary Research Communications. 24 (2000), 371-377.
Topiæ Popoviæ, Natalija; Hacmanjek, Mato; Teskeredžiæ, Emin: Health status of rudd ( Scardinius erythrophtalmus hesperidicus H.) living in the lake Vrana on the island of Cres. Journal of Applied Ichthyology. 17 (2001), 1, 43-45.
Èož-Rakovac, Rozelindra; Strunjak-Peroviæ Ivanèica; Topiæ Popoviæ, Natalija; Hacmanjek Mato; Šimpraga Borka, Teskeredžiæ Emin: Health status of wild and cultured sea bass in the northern Adriatic Sea. Vet. Med.-Czech, 47 (2002), 8: 222-226.
6. Equipment and Facilities Plan
All current activities are conducted on equipment temporarily provided by our research collaborators and the Rudjer Boskovic Institute. As detailed in Section 6.1, to set-up and implement the proposed Center, certain equipment is required.
6.1. Highly needed equipment for initial activities
Field vehicle $ 20,000
Biochemical blood analyzer (VETTEST 8008)$ 12,500
Mini-centrifuge (StatSpin)$ 2,000
Laboratory microscope 2000x magnification with image analyzer$ 19,000
Laminar flow hoods$ 13,000
Freezer and refrigerating units$ 10,000
Thermostat$ 1,000
API rapid kits for bacterial identification (Biomerieux)$ 3,000
Liquid nitrogen container$ 2,500
Water quality probe unit$ 2,300
Electrophoresis units$ 4,000
Western blotting kits$ 3,000
Spectrophotometer$ 3,500
Total:$ 95,800
7. Organizational Plan
A plan is to coordinate scientific and technical professionals in order to implement our above stated missions. Additional staff will be hired according to the need of the current projects. Gradually, the number of active researchers in diagnostics of aquatic animal diseases will increase.