05. MARACEK,I.: Research and expoitation of biotechnolgies in veterinary medicine. (Biotechnológiai és biotechnikai K+F program Szlovákiában)
05. Maracek
érk: 97.11.25.
UEVM, 04001 Kosice, Hlinkova 1/A, Slovak Republic
RESEARCH AND EXPLOITATION OF BIOTECHNOLOGIES IN VETERINARY MEDICINE
The main objectives of the STP
The development of increased potency probiotics for utilization in prevention and therapy of digestive disturbances, diarrhoea of the young, secretion disorders and mammary gland inflammations, puerperium endometritis and also for supporting defence mechanisms and the immune system.
Development and elaboration of new diagnostic kits based on molecular-genetic tests (PCR, RFLP) and monoclonal antibodies for diagnostics of rabies, feline AIDS, pestiviruses (CSFV, BVDV, IBR, BDV and anaerobic pathogens (Cl. perfringens).
Development of ELISA diagnostic methods based on monoclonal antibodies against staphylococcus enterotoxins A, B, C and D.
Preparation of DNA primers for diagnostics of enterotoxigenic strains of S. aureus.
Innovation and improvement of the effectiveness of serodiagnostics of most serious animal parasites, genetic classification of Lyme boreliosis and development of the method of target transport of anthelminth preparations in an animal organism.
Innovation and improvement of methods of controlled reproduction applying biotechnological products, particularly molecular immunology.
Development of biotechnological procedures of ensuring good health state, reproduction and production of ungulates, pheasants and wild boars.
Innovation of biotechnological methods of treatment of secondary raw materials to ensure their hygienic recycling and proposal of new technology of processing of poultry dung by means of coprophagous insect.
Development of biological-molecular methods of detection of antimicrobials, animal proteins and new pathogens in milk.
Development of a model of in vitro testing of toxicity using a tissue culture method.
Development of technological procedure and introduction of production of microbial stains and their application in the food industry.
Development of new immunodiagnostic procedures in transmissive spongiform neuropathies.
Development of a method of economic evaluation of biotechnological procedures in prevention of diseases and ensuring good health state of animals, safe products and sound productive environment.
The main expected achievements of the STP project in terms of practical exploitation:
The proposal of system approaches concerning the improvement of health of farm animal herds with regard to the freedom from serious infections and infestation.
New preparations and procedures intended to potentiate the mucosal defence abilities of the digestive tract of the young, udder and endometrium of dairy cows.
The development of an original ELISA method for detection of the virus of classical swine fever in the blood of animals.
Utilization of polymerase chain reaction and hybridization of nucleic acids in detection and differentiation of pestiviruses in comparison with analysis of their antigenic structure by means of monoclonal antibodies.
Preparation of monoclonal antibodies against staphylococcus enterotoxins C, D, and ELISA tests for determination of enterotoxins A, B, C and D.
Phylogenetic determination of isolated pestiviruses by means of PCR with epizootiological utilization.
Development of a system for monitoring of slow viral infections, including prionoses as etiological agents of spongiform encefalopathies in animals (sheep scrapie, BSE and similar).
Innovation of diagnostic methods and procedures for detection of anaerobes in animals and anaerobic microbial contamination of their products, i.e. food and raw materials of animal origin.
Preparation of an effective and harmless vaccine against fowl rheoviral infections.
Methodical procedures for elimination and compensation of factors which exert a negative influence on fertility and reproductive parameters.
Biotechnical methods of systemic character for optimization of controlled reproduction of farm animals within the scope of veterinary programmes oriented on good health state of herds, breeding, farms, etc.
New ELISA and RIA tests for diagnostic examination of milk samples substituting tests in animal blood samples.
New technologies of recycling the biogenic substances present in wastes which can serve as secondary raw materials. This concerns wastes of lignin-cellulose character as well as those from food processing industry or excrements of some animal species (poultry).
The structure of VTP:
Partial task 01:
"Application of probiotics and microbiotics in ensuring good health state of farm animals and its protection" (10 subject stages)
Team leader:
Alojz Bomba, DVM, PhD., Research Institute of Experimental Veterinary Medicine, Košice
Objectives of the research:
- a) Development of increased-potency probiotics for prevention and therapy of diarrhoea in the young, elimination of digestive disturbances and stimulation of the immune system
- b) Development of preparations based on lactobacilli for stimulation of natural mechanisms in the mammary gland, prevention and therapy of puerperium endometritis, promotion of conservation of green bulk feed and prevention of serious diseases of productive bee communities
- c) Development of a method for verification of the effect of biotechnology products on the programmed death of immune system cells
- d) Establishment of a biotechnological centre for the development, verification and production of probiotic preparations intended for young farm animals
Partial task 02:
A9"Utilization of biotechnology products and application of methods of molecular biology in diagnostics and immunoprophylaxis of infectious diseases in animals including zoonoses (8 subject stages)
Team leader:
Assoc.Prof.Stefan Svrcek, DVM, PhD, University of Veterinary Medicine and Research Institute of Experimental Veterinary Medicine, Kosice
Objectives of the research:
- a) Development of laboratory procedures for diagnostics of rabies and othe lyssa-viral diseases using in vitro methods and biotechnology products (monoclonal antibodies, PCR)
- b) Development of methodical procedures for diagnostics, prevention and suppression of feline AIDS
- c) Development of a molecular-genetic test (PCR) for diagnostics of pestiviruses
- d) Development of ELISA methods for diagnostics of BVD
- e) Innovation of diagnostic methods (PCR) of anaerobic pathogens in foodstuffs and animal feed
- f) Biotechnological construction and exploitation of biologically active preparations against actinobacillus pleuropneumoniae of pigs
- g) Development of new vaccines against rheovirus fowl infections and production of diagnostics preparations
Partial task 03:
"Molecular-genetic methods of detection of enterotoxins-producing staphylococci, determination of enterotoxins and some hormones for use in diagnostics" (3 subject stages)
Team leader:
Research Institute of Experimental Veterinary Medicine, Koaice
Objectives of the research:
- a) Preparation of monoclonal antibodies against staphylococci enterotoxins C, D and progesterone
- b) Development of an ELISA test for determination of staphylococci enterotoxins A, B, C and D
- c) Preparation of DNA primers for diagnostics of enterotoxigenic strains of Staphylococcus aureus
Partial task 04:
"The most important parasitoses in farm animals, their diagnostics, prevention and therapy using new biotechnological procedures" (5 subject stages)
Team leader:
Assoc. Prof. Pavol Dubinský, DVM, DSc., Parasitological Institute of SAS, Košice
Objectives of the research:
- a) Innovation and increasing the effectiveness of serodiagnostics of most important animal parasitoses
- b) Genetic classification of Lyme boreliosis agents and obtaining an antigen for production of an immunodiagnostic kit
- c) Determination of specific immunological characteristics of parasites for their reliable immunodiagnostics and differentiation
- d) Development of methods of target transport of antihelminth active ingredients with the aim to decrease the doses of medicals and their residua in tissues
Partial task 05:
"Biotechnological and biotechnical procedures in ensuring and protecting the productive health of farm and domestic animals" (9 subject stages)
Team leader:
Assoc. Prof. Alexander Michna, DVM, PhD., University of Veterinary Medicine, Košice
Objectives of the research:
- a) Development of a proposal for elimination of negative effects of magnesite light ashes on the health state and production of farm animals reared in exposed areas
- b) Investigation and affecting mucosal immunity of the young of farm animals by means of feed additives based on microbiotics
- b) Verification of the possibility of utilization of some enzymes, probiotics and biostimulators for protection of health and improvement of productive parameters in fowl
- d) Application of PNMK produced by biotechnology method as preventive and therapeutic means for domestic animals
- e) Introduction of an undemanding and reliable method of determination of amino- nitrogen in free amino acids for quantitative measurement of their concentration in samples of animal origin.
Partial task 06:
"Biotechnological and biotechnical procedures in the control of reproductive processes of farm animals and checking their genetic health" (7 subject stages)
Team leader: Eva Bekeová, DVM, PhD., Research Institute of Experimental Veterinary Medicine, Košice
Objectives of the research:
- a) Research and development of new biotechnical and biotechnological methods for controlled reproduction in small ruminants with gradual replacement of synthetic steroids exhibiting long protective period
- b) Improvement of biotechnics of obtaining cumulo-oocyte complexes, their conservation, in vitro maturation, fertilization, additional cultivation and transfer in ruminants
- c) Innovation of biotechnological methods for in vitro production of bovine pre- implantation embrya usable for cattle selection
- d) Innovation of biotechnical methods in the system of controlled reproduction in sheep and pigs
- e) Introduction and utilization of biotechnological methods in checking the genetic health of animals
Partial task 07:
"Biotechnological and biotechnical procedures affecting health, reproduction and production of game" (4 subject stages)
Team leader:
Assoc. Prof. Eng. Juraj Ciberej, PhD., University of Veterinary Medicine, Košice
Objectives of the research:
- a) Development of biotechnological and biotechnical procedures for ensuring the health, reproduction and production of ungulates
- b) Development of biotechnology of artificial rearing of pheasants from the point of view of improvement of their life span in the wild
- c) Development of new generation vaccines using biotechnological methods and analysis of epizootiological situation of pseudorabies in wild boar populations
Partial task 08:
"Research and utilization of biotechnological methods in ensuring hygiene-safety of foodstuffs and their technological treatment" (5 subject stages)
Team leader:
Eva Beliková, DVM, PhD, Research Institute of Experimental Veterinary Medicine, Kosice
Objectives of the research:
- a) Development of screening methods for detection of animal proteins of ruminants in thermally processes products, feed and canned goods
- b) Development of molecular-biological methods for determination of antimicrobial substances in milk
- c) Development of a suitable model for testing the toxic effects of chemicals using tissue cultures
- d) Development of a technological procedure and launching production of microbial stains and their utilization in the food industry
Partial task 09:
Hygienization of secondary raw materials from animal production and food processing industry" (4 subject stages)
Team leader:
Ján Venglovský, DVM, PhD., Research Institute of Experimental Veterinary Medicine, Košice
Objectives of the research:
- a) Innovation and modification of biotechnological methods of treatment of secondary raw materials from animal production and food processing industry for the purpose of their hygienic recycling
- b) Development of procedures of elimination of odourous substances present in the air in animal houses by means of biotechnological methods
- c) Verification of most suitable procedures for affecting the nitrogen metabolism and metabolism of organic substances in the process of treatment and processing of secondary raw materials
Partial task 10:
"Biotechnological processing of fowl excrements" (8 subject stages)
Team leader:
Prof. K. Boda, DVM, DSc., Research Institute of Experimental Veterinary Medicine, Košice
Objectives of the research:
- a) Development of a proposal for technology of fowl excrement processing by means of coprophagous insect
- b) Development of a technical procedure for utilization of biohumus obtained from fowl dung for manuring field crops and utilization of fly larvae and pupae in the nutrition of fowl, aquarium fish and exotic birds
- c) Development of non-standard equipment for biotechnological processing of fowl dung under operational conditions
- d) Development of technological set-up for an operation of capacity 1 to 2 t of excrements per day
- e) Determination of basic parameters of large-capacity technology for processing of fowl excrements, of capacity 5 - 10 t and 10 - 20 t per day
Partial task 11:
"Transmissive spongiform encefalopathies - lethal diseases transmissible from animals to man" (8 subject stages)
Team leader:
Assoc. Prof. Jozef Sokol, DSc., State Veterinary Administration of SR, Bratislava, and Prof. MVDr. Michal Novák, DrSc., Neuroimmunological Institute of SAS, Bratislava
Objectives of the research:
- a) Development of new immunodiagnostic procedures for transmissive spongiform neuropathies
- b) Pilot plant production of immunodiagnostic and genotypization kits and their introduction to laboratory diagnostics
- c) Identification, cloning and stabilization of hybridomas producing specific monoclonal antibodies against prion proteins of farm and domestic animals
- d) Development of a rapid method for isolation of DNA from the peripheral blood of sheep
- e) Development of screening tests for identification of positive hybridomas producing monoclonal antibodies to CNS
- f) Development of a kit for amplification and visualisation of PrP DNA
Partial task 12:
"Research and verification of possibilities of analysis and evaluation of economics of farm animals health" (3 subject stages)
Team leader:
Eng. A. Havrila, M.Agr.Sc., Research Institute of Experimental Veterinary Medicine, Košice
Objectives of the research:
- a) Social-economical evaluation of biotechnological methods and procedures in the sphere of animal health, animal products and environment
- b) Economics of the progress in utilization of biotechnologies in veterinary medicine
- c) Establishment of a database of information sources in the field of biotechnologies in veterinary medicine and access to this database by means of INTERNET
*
BIOTECHNOLÓGIAI ÉS BIOTECHNIKAI K + F PROGRAM SZLOVÁKIÁBAN
A program fö céljainak megoldása:
A potenciális probiotikumok kifejlesztése az emésztési zavarok megelőzésére, hasmenéses növendékállat-betegségek gyógyítására, kiválasztási zavarok, tejmirigy-gyulladás, szülés utáni méhnyálkahártya gyulladás megszûntetésére, és az ellenállóképesség mûködésének elősegítésére az immunrendszer révén.
Laboratóriumi diagnosztikai kitek kidolgozása molekuláris tesztekkel (PCR, RFLP) és monoklonális ellenanyagok alkalmazásával a veszettség diagnosztikájában, a macska-AIDS, pestisvirusok (CSFV, BVDV, IBR, BDV) és anaerob patogének (Cl.perfringens) terén.
ELISA diagnosztikai módszerek alkalmazása a Staphylococcus A, B, C és D-enterotoxin ellen.
DNS-primerek készítése enterotoxinogén St.aureus törzsek diagnosztizálására.
Paraziták szerológiai diagnosztizálásának megújítás a gén klasszifikálása (a Lym-féle borreliózisnál alkalmazott módszerek kidolgozása) által, a helminteknek az állati szervezetben való transportja alapján.
Az irányított (tervezett) szaporítás fokozása biotechnológiai, főleg molekuláris immunológiai módszerek alkalmazása révén.
Biotechnológiai eljárások kidolgozása a másodlagos termékek egészségileg nem kifogásolt reciklizálása érdekében, új technológiai eljárások kidolgozása a tejben található patogének antimikrobás anyagok által történő eliminálása érdekében.
Toxicitás tesztek modelljének kidolgozása in vitro módszerekkel, szövet kultúrákon.
Technológiai eljárás kidolgozása mikrobiális festékeknek az élelmiszeriparban történő bevezetésére.
Új immundiagnosztikai eljárás kidolgozása a transzmisszív spongioform neuropátia terén.
A gazdasági értékelés módszerének létrehozása biotechnológiai eljárások hasznosítása terén az állatok egészségvédelmében és állati termékek termelésében.
A K + F program várható fő eredményei:
Rendszeres javaslatok a tenyésztállatok gyógyítása, gazdasági állatok járványos és inváziós betegségektől való mentesítése terén.
Új eszközök és eljárások növendékállatok bélnyálkahártyája védekezőképességének erősítésében, fejőstehenek endometritiszének kezelésében.
Eredeti ELISA teszt kialakítása a klasszikus sertéspestísvírus kimutatására az állatok véréből.
Polimeráz láncreakció és nukleinsav szondák felhasználása a pestisvírusok megkülön-böztetésére, monoklonális ellenanyagok elemzése útján.
Monoklonális ellenanyagok előállitása a Staphylococcus enterotoxin C, D-re és ELISA tesztek az A-, B-, C- és D-enterotoxinok meghatározására.
A pestisvírusok filogenetikai izolálásának a meghatározása PCR-rel.
Lassú vírusos infekciók vizsgálati rendszerének kialakítása beleszámítva a prionokat, mint etiológiai tényezőket az állatok spongiform enkefalopátiáinál (paraplegia enzootica oviec-klusavka, BSE).
Diagnosztikai módszerek felújítása anaerobok kimutatására állatokban és állati termé-kekben, különös tekintettel az élelmiszerekre és állati eredetû takarmányokra.
Hatásos és nemkáros vakcína elöállítása a baromfi reovírusos fertőzése ellen.
Módszeres eljárások olyan tényezők csökkentésére, melyek negatívan hatnak a termé-kenység és szaporodás mutatóira.
Biotechnikai módszerek a szaporodás tervszerû optimalizálása gazdasági állatállomá-nyokban, tenyészetekben, farmokban.
Új ELISA és RIA tesztek a tej diagnosztikai vizsgálatára a vér helyett.
Lignocellulóz jellegû anyagok, élelmiszeripari hulladékok, vagy állati (baromfi) ürülé-kek, mint biogén anyagok reciklizáló technológiáinak kidolgozása.
A K+F 12 részfeladatból tevődik össze:
01. Probiotikumok és mikrobiotikumok érvényesítése a gazdasági állatok tenyésztésében és védelmében.
02. Biotechnológiai termékek felhasználása a diagnosztikában és immunprofilaxisban a fertőző betegségeknél, beleszámítva a zoonózisokat.
03.Molekuláris genetikai módszerek a Staphylococcus törzsek enterotoxinjainak elő-állításában, kimutatásában és némely hormonok meghatározásának diagnosztikai felhasználásában.
04. A legjelentősebb állati (élősködök) paraziták, diagnosztikája, gyógyítása új biotech-nológiai eljárásokkal.
05. Biotechnológiai és biotechnikai eljárások a termelésben, a gazdasági- és háziállatok egészségvédelmében.
06. Biotechnológiai és biotechnikai eljárások a gazdasági állatok szaporodási folyama-taiban és a genetikai egészség ellenőrzésében.
07. Biotechnológiai és biotechnikai eljárások hatása az egészségre és szaporodásra a vadgazdaságban.
08. Egészségileg nem kifogásolt biotechnológiai módszerek alkalmazása az élelmiszer-iparban és módosítot gyártásban.
09.Másodlagos állattenyésztési és feldolgozó ipari termékek ártalommentesítése.
10.Baromfi exkrementek (ürülék) biotechnológiai feldolgozás.
11. Állatról emberre terjedő transzmisszív spongioform letális enkefalopátiák vizsgálata.
12. Kutatások és lehetőségek hitelesítése, ökonómiai értékelése.
Execution of VTP research
Research capacity in man-years (FTE)
1997 1998 1999 Total
85.7 179.9 205.3 470.9