Kazakh National agrarian university UDC: 636.087.63:639.2 On the rights of manuscript PARITOVA ASSEL ERZHANOVNA Veterinary-sanitary evaluation of the quality fish while using non- traditional feed additives «Tseofish» 6D120200-Veterinary sanitation A dissertation submitted for the degree of Doctor of Philosophy (PhD) The domestic scientific adviser doctor of veterinary sciences, professor Sarsembayeva N.B.; Foreign scientific adviser doctor of chemical sciences, professor of Institute Plant protection (Poland) Lozowicka B. The Republic of Kazakhstan Almaty, 2014 1 CONTENTS NORMATIVE LINKS 4 DEFINITIONS 6 DESIGNATIONS AND ABBREVIATIONS 8 INTRODUCTION 10 1 REVIEW OF LITERATURE 14 1.1 Veterinary-sanitary examination of fishery products 14 1.2 Aquaculture development in Kazakhstan and actual problems 18 1.3 Feed and feed additives in aquaculture 20 1.4 Zeolites are natural minerals used in veterinary science 23 1.5 The chemical composition and nutritional value of fish 30 1.6 Pesticide residues in feed and fish 35 2 OWN RESEARCHES 38 2.1 Materials and methods 38 2.1.1 Research materials 38 2.1.2 Research methods 41 2.1.2.1 Organoleptic methods 41 2.1.2.2 Methods of biochemical research 42 2.1.2.3 Methods for determining of chemical composition of fish 43 2.1.2.4 Method for determining the amino acid composition of fish 43 2.1.2.5 Methods of determining fatty acid composition of fish 43 2.1.2.6 Method for determining the mineral composition of fish 44 2.1.2.7 Method for determining the vitamin composition of fish 44 2.1.2.7.1 Equipment 44 2.1.2.7.2 Reagents and solutions 44 2.1.2.7.3 Samples and reference materials 44 2.1.2.7.4 Sample preparation 45 2.1.2.7.5 Chromatographic conditions 45 2.1.2.8. Methods of histological examination of meat 45 2.1.2.8.1 Preparing a mixture of egg white with glycerol and processing 45 microscope slides 2.1.2.8.2 Preparing of eosin solution 46 2.1.2.8.3 Preparation of Ehrlich hematoxylin 46 2.1.2.8.4 Staining of slices 46 2.1.2.9 Hematological study of fish 46 2.1.2.10 Determination of pesticide residues in feed and fish meat 48 2.1.2.10.1 Samples and reagents 48 2.1.2.10.2 Standarts 48 2.1.2.10.3 Sample preparation 48 2.1.2.10.4 Instrumentation and chromatographic conditions 48 2.1.2.10.5 Method of validation 49 2.1.2.11 Statistical processing of results 50 3 RESEARCH RESULTS 51 2 3.1 Physico-chemical properties of non-traditional feed additive Tseofish 51 3.2 Toxicological evaluation of non-traditional feed additive Tseofish 55 3.3 Study of the effect of non-traditional feed additive Tseofish at fish- 56 biological and hematological parameters of the body of fish 3.3.1 Fish breeding and biological indicators of valuable fish species when 56 using nontraditional feed additive Tseofish 3.3.2 Hematologic characteristics of valuable fish species when using a feed 58 additive Tseofish 3.4Veterinary and sanitary assessment of the quality of fish, when used in 63 the composition of the feed additive feed Tseofish 3.4.1 Organoleptic characteristics and indicators of freshness of fish meat in 63 the diet added feed additive Tseofish 3.4.2 The chemical composition and nutritional value of fish meat when 70 using a feed additive Tseofish 3.4.3 Amino acid composition of fish meat in the diet when using NFA 75 Tseofish 3.4.4 Analysis of the fatty acid composition of fish meat when added to feed 81 NFA Tseofish 3.4.5 Vitamin content of fish meat of valuable species when used in feed 93 NFA Tseofish 3.4.6 Mineral content of fish meat in the application of NFA Tseofish 97 3.5 Morphological changes of valuable species of fish meat using NFA 100 Tseofish 3.6 Study of the content of pesticide residues in feed and fish 102 3.6.1 Pesticide residues in fish feeds when using nontraditional feed additive 117 Tseofish 3.6.2 Pesticide residues in fish when using in feed nontraditional feed 102 additive Tseofish 4 GENERALIZATION AND EVALUATION OF RESEARCH 120 RESULTS 5 CONCLUSION 123 6 PRACTICAL PROPOSALS 125 7 REFERENCES 126 Application A Pesticide residue chromatograms Application B The electron fotos of zeolite crystals Application C Act of research results in the learning process 3 NORMATIVE LINKS In this dissertation references are used to the following standards: ST RK 1802-2008 Fish, seafood and products their processing. Acceptance rules and sampling. ST RK 1803-2008 Fish and seafood. Sensory evaluation methods. GOST 7631-85 Fish, marine mammals, marine invertebrates and products their processing. Acceptance rules, organoleptic methods of quality evaluation, sampling methods for laboratory testing. GOST 23042-86 Meat and meat products. Method for determination of total fat content. GOST R 51479-99 Meat and meat products. Method for determination moisture mass fraction. GOST 23392-78 Meat. Methods for chemical and microbiological analysis of freshness. GOST 19496-93 Meat. Method of histological analysis. GOST R 53642-2009 Meat and meat products. Method for determination of total ash mass fraction. GOST 25011-81 Meat and meat products. Methods for determination of protein. ST RK 2010-2011 Determination of organochlorine pesticides by chromatographic methods. ISO 24333-2009 Grain and grain products. Sampling. GOST 25336-82 Laboratory glassware and equipments. Types, basic parameters and dimensions. GOST 13496.20-87 Fodder and feed raw materials. Method for the determination of pesticide residues. The Council Directive Veterinary and sanitary requirements for aquaculture 2006/88/ES from animals and their products, and on the prevention and October 24, 2006 control defined diseases in aquaculture animals. GOST R 52346-2005 Combined feed for fish. Nomenclature of indices. ISO/IEC 17025:2005 General requirements for the competence of testing and calibration laboratories. ISO/TC 34/SC 6 Meat, poultry, fish, eggs and their products. ISO 936:1998 Meat and meat products - Determination of total ash. ISO 1442:1997 Meat and meat products - Determination of moisture content (Reference method). ISO 12877:2011 Traceability of finfish products. Specification on the information to be recorded in farmed finfish distribution chains. CAC/MRL 1 Maximum Residue Limits (MRLs) for Pesticides. 4 CAC/GL 55-2005 Guidelines for Vitamin and Mineral Food Supplements. CAC/GL 33-1999 Recommended Methods of Sampling for Pesticide Residues for the Determination of Compliance with MRLs. CAC/RCP 52-2003 Code of Practice for Fish and Fishery Products. CAC/GL 31-1999 Guidelines for the Sensory Evaluation of Fish and Shellfish in Laboratories. GOST R 55483-2013 Meat and meat products. Determination of fatty acid composition by gas chromatography. GOST R 55482-2013 Meat and meat products. Method for determination of water-soluble vitamins. 5 DESIGNATIONS AND ABBREVIATIONS ADIs- Acceptable Daily Intakes ARfDs -Acute Reference Doses ASPM -Agreement about Sanitary and Phytosanitary measures ATP -Adenosine triphosphate BD - basic diet DDT - 4,4 '-dichlorodiphenyl-trichloroethane E.coli - Esherichia coli ECD - electron capture detector EFA - essential fatty acids FA- fatty acid FAME - fatty acid methyl ester FAO -Food and Agriculture Organization GOST - government standard GC - a gas chromatograph g - gram g/l - gram per litre h - hour HAC- Higher Attestation Comission HCH - hexachlorocyclohexane EU - European Union EUPT - European Commission’s Proficiency Testing Program ISO - The International Organization of Standartization IUCN - The International Union for Conservation of Nature kg - kilogram lb - the pound-mass LD - lethal dose LIA - lioneic acid LOD - limit of detection LOQ - limit of quantification mcl - microliter mg - milligram mg/kg- milligram/kilogram ml- milliliter mm - millimeter min - minute MPC - maximum permissible concentration MRL - maximum permissible level MYA - myristic acid ND - normative documents NFA - nontraditional feed additive NPD - the nitrogen-phosphorus detector OCs- organochlorinies OLA- oleic acid 6 PAA- palmitic acid PUFA - polyunsaturated fatty acid RfD – Reference Dose RSD - relative standard deviation S/N - signal-to-noise ratios SRI - Scientific Research Institute STA- stearic acid ST RK - Standard of the Republic of Kazakhstan UK - United Kingdom USA- United States of America WTO - World Trade Organization 7 DEFINITIONS This dissertation uses the following terms with corresponding definitions: Adsorption - a process that occurs when a gas or liquid solute accumulates on the surface of a solid or a liquid (adsorbent), forming a molecular or atomic film (the adsorbate). It is different from absorption, in which a substance diffuses into a liquid or solid to form a solution. The term sorption encompasses both processes, while desorptionis the reverse process. A fish - is any member of a paraphyletic group of organisms that consist of all gill-bearing aquatic craniate animals that lack limbs with digits. Aluminosilicates - are minerals composed of aluminium, silicon, and oxygen, plus countercations. They are a major component of kaolin and other clay minerals. A mineral - is a naturally occurring substance that is solid and stable at room temperature, representable by a chemical formula, usually abiogenic, and has an ordered atomic structure. Amino acids - are biologically important organic compounds composed of amine (-NH2) and carboxylic acid (-COOH) functional groups, along with a side- chain specific to each amino acid. A vitamin - is an organic compound required by an organism as a vital nutrient in limited amounts. An organic chemical compound (or related set of compounds) is called a vitamin when it cannot be synthesized in sufficient quantities by an organism, and must be obtained from the diet. Chromatography - (kroʊməˈtɒɡrəfi/; from Greek χρῶμα chroma "color" and γράφειν graphein "to write") is the collective term for a set of laboratory techniques for the separation of mixtures.