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Генеральный директор Международного эпизоотического бюро (МЭБ) Бернар Валла ознакомился с работой ФГБУ «ВГНКИ»

07.03.2013

В рамках подписания Соглашения об открытии в Москве регионального представительства Международного эпизоотического бюро (МЭБ) его генеральный директор Бернар Валла ознакомился с работой подведомственного Россельхознадзору «Российского государственного Центра качества и стандартизации лекарственных средств для животных и кормов» (ФГБУ «ВГНКИ»). Ему были продемонстрированы презентации двух докладов, характеризующих работу двух отделов института - отдела безопасности кормов и пищевых продуктов (заведующий – Комаров А.А., д.б.н., профессор) и отдела биотехнологии (заведующий - Обухов И. Л., д.б.н., профессор).  Приводим тексты презентаций.

Пресс-служба ФГБУ «ВГНКИ»
 
Презентация Отдела безопасности кормов и пищевых продуктов (Department of feed and food products safety). Заведующий отделом — доктор биологических наук, профессор Комаров А.А.
 Main Tasks of Department
  1. Development and validation of new confirmatory and screening methods for control of drug residues and toxic substances in animal produce.
  2. Monitoring of products of animal origin in the frames of Government Monitoring Program.
  3. Testing of feed additives and animal drugs for registration and certification in Russia.
  4. Expert assessment of normative documents for feed additives and animal drugs
  5. International collaboration. Participation in International Proficiency Testing.
  6. Organizing Proficiency Tests for Inter-regional Reference Centers of Federal Service for Veterinary and Phytosanitary Surveillance
       Instrumentation
       2 HPLC – triple quad systems
       2 HPLC – hybrid triple quad/ ion trap systems
       2 UPLC – hybrid Q-TOF HR MS system
       1 GC- HR MS system with double-focusing geometry
       1 GC – triple quad system
       1 GC – ion trap system
       HPLC – ICP MS system
       3 stand-alone HPLC instruments
       2 Atomic Absorption Spectrometers
       Spectrophotometers
       Different equipment for sample preparation
       Test assays developed by VGNKI for screening of drug residues
16 test assays developed
       Test assay «Clenbuterol-ELISA»
       Test assay «Methyltestosteron-ELISA»
       Test assay «19-nortestosteron-ELISA»
       Test assay «Diethylstilbestrol-ELISA»
       Test assay «Trenbolon-ELISA»
       Test assay «Ethynilestradiol-ELISA»
       Test assay «Dexamethasone-ELISA»
       Test assay «Chloramphenicol-ELISA»
       Test assay «Tetracyclines-ELISA»
       Test assay «Quinolones-ELISA»
       Test assay «Enrofloxacine-ELISA»
       Test assay «Malachite green-ELISA»
       Test assay «Zearalenone-FPIA»
       Test assay «DON-FPIA»
       Test assay «Ochratoxin A-FPIA»
       Test assay «Aflatoxin B1-FPIA»
       Use of confirmatory multi-methods to increase the testing efficiency
       Confirmatory methods of drug residues detection are developed on the basis of a combination of methods GС-MS/MS (Gas Chromatography-Mass Spectrometry), HPLC-MS/MS (High Performance Liquid Chromatography-Mass Spectrometry) (methods of I level on Codex Alimentarius).
       New perspective direction is development of multi-methods that allow to carry out simultaneous detection of many xenobiotics in a sample and essentially simplifies a problem of carrying out a large-scale monitoring and increases efficiency of techniques.
       Application of HPLC-MS with tandem mass spectrometer detection allows to carry out reliable detection of several groups of the various chemical nature in one sample with the minimum preparation of the sample.
       We have optimized conditions for quantitative determination of more than 200 drugs, growth promotors and their metabolites in food of animal origin
       Monitoring of foods of animal origin in 2012
       Determination of tetracycline residues
       Tetracyclines are antibiotics most often used in livestock
       Wide-spread usage of tetracycline antibiotic group, and also long period of excretion (more than 20 days) increase the probability of residues in products of animal origin
       The confirmatory method, allowing to define reliably antibiotic residues of tetracycline group in products of animal origin with HPLC-MS (High Performance Liquid Chromatography – Mass Spectrometry), was developed by “VGNKI”
       Doxycycline MRL (SanRules 2.3.2.2804-10) is harmonized with EU requirements - ЕС (Commission Reg. No 37/2010)
       VGNKI’s method is approved as national standard: “GOST R 53601-2009”
       Occurrence of samples with exceeding MRL for tetracyclines in 2008-2012
      
Occurrence of tetracyclines residues in food
from different countries in 2011-2012
       Occurrence of coccidiostats residues in food in 2012
       Detection of not only drug residues in production of animal origin but also their metabolites
       Nitrofurans are quickly metabolized in the organism, however their residues stay in organs and tissues of productive animals for a long time (more than 6 weeks after application)
       Nitrofuran metabolites possess carcinogenic, mutagen and genetic toxic actions, so no MRL or ADI could be established.
       Any nitrofuran residues in products of animal origin is prohibited in most countries!
       Use of preparations of this group is forbidden in animal industries of the USA, Canada, countries of the European Union, Argentina, Brazil, Australia and other countries!
       Since 01.01.2012г, according to SanRules 2.3.2.2871-11, MRL of 1 mcg/kg for nitrofurans is established 1 μg/kg, it practically eliminates the possibility legal use of nitrofuran drugs.
       VGNKI developed the national standard for determination on nitrofurans and metabolite – GOST R 53992-2010.
 
       OCCURRENCE OF NITROFURANS RESIDUES IN 2011-2012
       Identification of new risks in 2012
       Risks associated with use of beta-agonists
       β-agonists – are non-steroid compounds, but expressing similar effects with steroids. They have strong anabolic effect and induce the growing of muscle mass.
       In Russia β-agonists are not registered neither as feed additives nor veterinary drugs.
       The risk of beta-agonists hazard to human health caused the ban on their use in EU countries (96/22/ЕС).
      
DETERMINATION OF ϐ-AGONISTS RESIDUES IN FOOD
        Triphenylmethane dyes
       Triphenylmethane dyes (malachite green, crystal violet and their leuco- metabolites) express antiparasitic and antifungal action and are widely used in fish production in some countries of south-East Asia
       They are widely used as industrial dyes and due to this can accumulate in environment and contaminate fish and seafood
       The use of these dyes is prohibited in USA, Canada, EU due to their carcinogenic and genotoxic effects (21 CFR 589.1000)
        No residues in animal products are allowed!
Ø In Russia triphenylmethane dyes are not registered as veterinary drugs, thus no residues are allowed as well.
       According to EU legislation the testing labs should reach the limit of detection of 2 µg/kg to determine dyes (Commission Decision 2004/25/EC). VGNKI has developed and validated UPLC/MS-TOF method with LoD of 1,0 µg/kg.
       DETERMINATION OF TRIPHENYLMETHANE DYES IN FISH
       Applying the precautionary principle: organoarsenics
       In USA organoarsenic compounds are registered and could be used in livestock production. Roxarsone and arsanilic acid are the components of more than 60 feed additives used in USA.
       MRL for arsenic in raw meat established in USA is 0,5 mg/kg, five times higher than in Russia (0,1 mg/kg)
       In Russia the organoarsenics are not registered, and previous analytical methods were aimed to determination of total arsenic.
       In 2010-2011 VGNKI has performed the experiments showing the possibility of determination of organoarsenics, based on HPLC-ICP-MS
       The method was developed and validated in VGNKI.
       Confirmation of technical competence according to ISO/IEC 17025
       Confirmation of technical competence according to ISO/IEC 17025
       EXTERNAL ADVANCE TRAINING COURSES FOR STAFF
       Development of methods for determination of triphenylmethane dyes in aquaculture
         Screening:
*      We optimized the conditions to develop sensitive and specific ELISA kit for malachite green.
         Confirmation:
*      An original sample preparation was designed for determination of triphenylmethane dyes. Preliminary oxidation of leucometabolites let to lower LoD to 1 µg/kg.
*      The chromatography separation and MS identification were optimized on high resolution MS system Maxis (Bruker Daltonik).
*      The accuracy of the method was confirmed with successful participation in FAPAS international proficiency testing on determination of malachite green in fish.
       VGNKI’s result in proficiency testing on determination of malachite green in fish (FAPAS, Oct 2012г)
       Development of confirmatory method for determination of aminoglycoside residues
       FPIA kits
       Outcome of scientific work in 2012
3 methods were designed and validated
Ø  Method of confirmatory determination of aminoglycoside residues in food by HPLC-MS/MS;
Ø   Method of confirmatory determination of anabolic steroids and stilbene derivatives in feed, biological liquids, tissues by HPLC-MS/MS;
Ø Method of confirmatory determination of triphenylmethane dyes in fish and aquaculture products by UPLC-TOF/MS.
Regulatory documents (standards and instruction) were designed for 4 test kits for quantification of mycotoxins in feed and food products by fluorescent polarization immunoassay (FPIA) : «Aflatoxin В1 FPIA», «Ochratoxin А FPIA», «Zearalenon FPIA», «DON FPIA».
       PARTICIPATION IN INTERNATIONAL CONFERENCES, SEMINARS, WORKSHOPS
Staff members took part 20 international events, including:
Ø 12th International Congress of the European Association for Veterinary Pharmacology and Toxicology, Leeuwenhorst, Netherlands (A.Komarov);
Ø 60th ASMS Conference on Mass Spectrometry and Allied Topics, Vancouver, Canada (S.Calugaru);
Ø «EuroResidue VII», Egmond-an-Zee, Netherlands (A.Komarov, I.Nesterenko, I.Batov);
Ø III International conference «Feed-Efficacy-Claims-Mode of Action», Cologne, Germany (A.Komarov);
Ø 6th Nordic Conference On Plasma Spectrochemistry (A.Sereda); 
Ø 28th Asilomar Conference on Mass Spectrometry in Food Safety and Quality, Pacific Grove, CA (S.Calugaru);
Ø IV International Conference «Feed safety: methods and tasks», Beijing, China
Ø 10 scientific articles published
 
Презентация отдела биотехнологии (Departmentofbiotechnology) Заведующий отделом — доктор биологических наук, профессор Обухов И.Л.
 
State-run monitoring of feed and food products safety (determination of ruminant DNA and GMO in feed and food products).
 Development, testing and validation of molecular biology methods for detection and quantification of new transgenic lines in feed and food products.
Development, testing and validation of molecular biology methods for the diagnostics of bacterial and viral infections in animals.
PCR diagnostics of animal infectious diseases.
Monitoring the efficiency of domestic animals vaccination against rabies in connection with international requirements established in EC Regulation № 998/2003.
Microbiological and mycological control of feed and food products safety.
PCR testing for viral contaminants of veterinary vaccines and application of genetic analysis in vaccine safety and quality control.
The identification of animal components in feed and food products by molecular biology methods
The PCR tests for bovine, equine, ovine, chicken, rat, mink and dog DNA detection in feed and food products have been developed.
The sequencing of mitochondrial gene cytB has been used to identify relative animal species to prevent falsification in food samples.
The Real-time PCR test has been developed to differentiate 3 salmon species (Oncorhynchus gorbuscha, O. keta and O. nerka) in caviar and fish products.
Development and testing of a novel biochip technology
Real-time PCR tests for the diagnostics of bacterial and viral infections in animals
BOVIDAE
- Bovine leukemia virus
- Bovine Herpesvirus
- Bovine viral diarrhea virus
- Rotavirus, Coronavirus
- M. tuberculosis/bovis
- Brucella abortus
- Fusobacterium necrophorum
- Yersinia pseudotuberculosis, Yersinia enterocolitica
- Parainfluenza-3 virus 
Real-time PCR tests for the diagnostics of bacterial and viral infections in animals
  AVES
Chlamydophila psittaci
Salmonella spp.
Mycoplasma synoviae
Mycoplasma gallisepticum
Influenza A virus (H5, H7, H9)
Newcastle disease virus
Avian infectious bronchitis virus
Ornithobacterium rhinotracheale
Avian infectious laryngotracheitis virus
Avian metapneumovirus
 
PCR tests for the diagnostics of bacterial and viral infections in animals
  MULTIPLE SPECIES
Chlamydia spp.
Salmonella spp.
Mycoplasma spp.
Bacillus anthracis
Influenza A virus
Mycobacterium paratuberculosis
M. avium 
Leptospira interrogans
Listeria monocytogenes
Campylobacter jejuni
 Aleutian mink disease virus
Bordetella bronchiseptica
Toxoplasma gondii
Aeromonas sp.
 
Recent PCR tests that were developed in the department
- Real-time PCR test for the detection of Schmallenberg viral RNA in animal tissues, blood, serum and in disease vectors (mosquitos).
697 different samples were analyzed, viral RNA
was not detected.
- Real-time PCR test for the detection of Actinobacillus pleuropneumoniae DNA in bacterial cultures, swine nasal swabs, tracheobronchial washes, tonsils and lung tissues.
138 different samples were analyzed, 14.5% were positive
-Real-time PCR test for the detection of bovine herpes virus 1 (BHV1) DNA.
Allows to detect viral DNA in semen, tissues, swabs of not vaccinated animals or
of animals vaccinated with marker vaccines.
78 different samples were analyzed, 12.3% were positive.