RU2392618C1 - Method of animal killing product quality evaluation - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention refers to veterinary, particularly to veterinary examination. Method involves biosample derivation on the basis of phenylisothiocyanate solution in isopropyl alcohol, exposure to capillary zonal electrophoresis using work buffer based on β-cyclodextrin, detection of free amino acids subjected further to qualitative and quantitative analysis using water extraction of animal organs and tissues as biosample. Biosample is taken in advance into microcontainer in amount not exceeding 0.05 cm³ of fluid to which up to 0.1 cm³ of 0.1 M solution of sodium carbonate is added, stirred, and then at least 0.3 cm³ of phenylisotiocyanate solution in isopropyl alcohol is added and left to react for up to 40 minutes. After the content is dried in natural conditions, 0.5 cm³ or less of distilled water is added, stirred, poured into Eppendorf tube and centrifuged to remove gases and suspensions. Further the data of qualitative and quantitative analysis are compared to reference result of free amino acid concentration for clinically healthy animals, and product quality is determined. In case free acid concentration exceeds reference, killing products are considered contaminated with helminths.

EFFECT: fast and accurate determination of helminth contamination of killing products.

3 dwg, 14 tbl, 1 ex

Description

The invention relates to the field of veterinary medicine, in particular to veterinary examination.

The determination of free amino acids in blood serum by chromatography on paper is known [see K.V. Giri et al., 1953, F. Bode, 1955. Reference. Biochemical research methods in the clinic. / Edited by Professor A.A. Pokrovsky. M .: "Medicine", 1969, p. 85-91], where the method is based on the reaction of amino acids with ninhydrin in a slightly acidic medium, followed by the conversion of the obtained blue derivative into a stable copper derivative of orange color, having a maximum absorption at 530 mmk. Measurement of the optical density of the copper derivative (“dida”) allows quantitative determination of all amino acids, with the exception of proline and hydroxyproline, which otherwise react with ninhydrin.

, где А - количество микрограммов аминокислоты, содержащейся в 100 мл сыворотки; а - количество микрограммов аминокислоты в пробе, найденное по графику; с - общее количество экстракта в микрограммах; b - количество микролитров экстракта, взятое для хроматографирования; d - количество микролитров сыворотки, соответствующее общему количеству экстракта.The determination process includes: 1) obtaining a concentrated protein-free extract of blood serum according to the method of Boulanger and Bizerte; preliminary, powder is obtained from 2 ml of blood serum dried by lyophilization or in a vacuum desiccator; 2) obtaining one-dimensional chromatograms of blood serum; 3) quantitative determination of free amino acids of blood serum; 4) the construction of a calibration graph. The calculation of the amino acid content is carried out according to the calibration graphs obtained with standard solutions of amino acids by the formula:

where A is the number of micrograms of the amino acid contained in 100 ml of serum; a - the number of micrograms of amino acids in the sample, found according to the schedule; C is the total amount of extract in micrograms; b is the number of microliters of the extract taken for chromatography; d is the number of serum microliters corresponding to the total amount of extract.

The closest in technical essence is the method for determining the free forms of amino acids in the blood serum of animals through the system of capillary electrophoresis "Drops" (see. Practical guidance on the use of capillary electrophoresis systems "Drops". N.V. Komarova, Ya.S. Kamentseva, S- P, 2008, p.102), including derivatization of a bioassay based on a solution of phenylisothiocyanate in isopropyl alcohol, exposure to it by capillary zone electrophoresis using a working buffer based on β-cyclodystrin, detection vobodnyh amino acids and their qualitative and quantitative analyzes.

The disadvantages of the known methods are the limited capabilities, the complexity of the study, the lengthy determination process, the inability to determine the quantitative composition of free amino acids in the extract of organs and tissues (the longest muscle of the back, heart muscle, liver, lungs, spleen and kidneys) with metastrongylosis (Metastrongylus elongatus) on quality products of slaughter of pigs in cattle with dicroceliosis (Dicrocoelium lanceatum) and echinococcosis (Echinococcus granulosus) on the quality of products of slaughter of cattle.

The technical solution to the problem is to expand the functional and technological capabilities, shorten the lead time and increase the accuracy of determining free amino acids in establishing the quality and safety of animal slaughter products in pig metastrongyles (Metastrongylus elongates); with dicroceliosis (Dicrocoelium lanceatum) and echinococcosis (Echinococcus granulosus) in cattle.

This object is achieved in that in a method for assessing the quality of animal slaughter products, including derivatization of a bioassay based on a solution of phenylisothiocyanate in isopropyl alcohol, exposure to it by capillary zone electrophoresis using a working buffer based on β-cyclodystrin, detection of free amino acids and their qualitative and quantitative analysis , according to the invention, a water sample of animal organs and tissues is used as a bioassay; preliminary, a bioassay is taken into a microvessel in a volume of no more 0.05 cm ³ of liquid, which is added to 0.1 cm ³ of 0.1 M sodium carbonate solution, stirred, and then added at least 0.3 cm ³ phenyl isothiocyanate solution in isopropyl alcohol and left to react to 40 minutes, how the contents dried in vivo, is not added more than 0.5 cm ³ of distilled water, stirred, poured into an Eppendorf tube and centrifuged to remove suspended solids and gases, and then the data obtained as a result of qualitative and quantitative analysis result is compared with a control to centration of free amino acids is clinically healthy animals and determine the quality of the products, if the concentration of free amino acids compared to control increases, the products of slaughter animals is considered infected with helminths.

The novelty of the proposed proposal is due to the fact that it ensures high separation of components, not available with other research methods; low consumption of reagents and solvents; simplicity of equipment; high speed analysis and high reproducibility of reaction conditions; good solubility of derivatives in an aqueous medium; commercial availability; wide possibilities of capillary electrophoresis in the analysis of free amino acids. In addition, the proposed proposal is more economical, because does not require the use of expensive chemicals.

The invention is illustrated by drawings, where figure 1 presents a graph of the concentration of free amino acids in the hood of lung tissue in pigs with metastrongylosis; figure 2 is a graph of the concentration of free amino acids in the extract of liver tissue in cattle with dicroceliosis; figure 3 is a graph of the concentration of free amino acids in the extract of liver tissue in cattle with echinococcosis.

A method for determining the quality of slaughter products of animals infected with helminths is as follows.

The method determines the concentration of free amino acids in a biological test, which is used as an aqueous extract of organs and tissues (the longest muscle of the back, heart muscle, liver, lungs, spleen and kidneys) in pig metastrongyles, dicroceliosis and cattle echinococcosis. The biological sample is preliminarily subjected to derivatization based on a solution of phenylisothiocyanate in isopropyl alcohol. For this, the biological sample is taken into a microvessel in a volume of not more than 0.05 cm ^{3 of} liquid, to which up to 0.1 cm ^{3 of a} 0.1 M sodium carbonate solution is added, mixed, then added not less than 0.3 cm ³ phenyl isothiocyanate solution in isopropyl alcohol and left to react to 40 minutes, after the contents dried in vivo, is not added more than 0.5 cm ³ of distilled water, stirred, poured into a test tube Eppend RF and centrifuged to remove gases and suspensions, then they are exposed to it by capillary zone electrophoresis using a working buffer based on β-cyclodystrin, then qualitative and quantitative analyzes for the detection of free amino acids are carried out and the data obtained as a result of qualitative and quantitative analyzes are compared with the control concentration result free amino acids of clinically healthy animals and determine the quality of products if the concentration of free amino acids is compared with the control increases, the animal slaughter products are considered substandard.

The use of capillary electrophoresis to determine the mass concentration of amino acids is based on the separation of the anionic forms of N-phenylthiocarbamyl derivatives of amino acids under the influence of an electric field due to their different electrophoretic mobility. To identify and quantify the analyzed components, ultraviolet absorption is recorded at a wavelength of 254 nm in the extract of organs and tissues (the longest muscle of the back, heart muscle, liver, lungs, spleen and kidneys) in pig metastrongyles, dicroceliosis and cattle echinococcosis in order to establish the effect metabolic products Metastrongylus elongates on the quality of pig slaughter products, Dicrocoelium lanceatum and Echinococcus granulosus - on the quality of cattle slaughter products.

An example of a specific implementation of the method for determining the quality of slaughter products of animals infected with helminths, for example, pigs infected with metastrongylus (Metastrongylus elongatus), and cattle infected with dicroceliosis (Dicrocoelium lanceatum) and echinococcosis (Echinococcus granulosus)

In the process of post-slaughter diagnostics of pigs in the amount of 500 animals in 100 animals, we were diagnosed based on the detection of a large number of helminths of metastrongilus in the lumen of the bronchi. In the process of post-mortem diagnosis in cattle in the amount of 2500 animals, among them 557 animals in the liver showed dicrocelia, 350 animals had echinococci in the liver and 150 animals in the lungs.

To determine the concentration of free amino acids in pig metastrongyles, dicroceliosis and echinococcosis of cattle, an organ and tissue extraction (the longest muscle of the back, heart muscle, liver, lungs, spleen and kidneys) is used. For this, for each disease of the animals, average samples are taken from 10 animals. In this case, the studied animals were divided into 2 groups of 10 average samples in each. The control group consisted of clinically healthy animals (pigs and cattle), and the experimental groups were affected by metastrongilus, dicrocelia, and echinococcus.

To determine the free amino acids in clinically healthy animals and in pig infestations, the causative agent of Metastrongylus elongatus, cattle - Dicrocoelium lanceatum, cattle - Echinococcus granulosus take a sample of 1.0 g of organs and tissues (the longest muscle of the back, heart muscle, liver, lungs , spleen and kidneys) and pour 5 cm ^{3 of} distilled water and insist 5-6 hours with periodic stirring. The content of free amino acids is determined through derivatives with phenylisothiocyanate by capillary electrophoresis.

0.05 cm ^{3 of} liquid is taken with a pipette into a microvessel (glass, bottle, etc.), 0.1 cm ^{3 of a} 0.1 M sodium carbonate solution is added, stirred, then 0.3 cm ³ of a phenyl isothiocyanate solution in isopropyl alcohol is added ( 0.2 cm ³ FITZ in 12 cm ³ isopropyl alcohol) and left to undergo a reaction for 40 minutes. After the contents have dried under natural conditions, 0.5 cm ^{3 of} distilled water is added, mixed, poured into an Eppendorf tube and centrifuged to remove gases and suspensions. The dilution in this case is 6. Next, an analysis is carried out.

For analysis, a Capill-103 R capillary electrophoresis device is used, which is equipped with an ultraviolet detector with a detector wavelength of 254 nm (for example, FIG. 1, FIG. 2 and FIG. 3), where a quartz capillary 0.5 m long to the detector is used , with an inner diameter of 75 × 10 ^-6 m; adjustable high voltage source of positive polarity 3-25 kV; hydrostatic injection of a sample under a pressure of 30 mbar for 5 sec; β-cyclodextrin-based buffer working solution (54 mg in 12.5 cm ^{3 of a} buffer solution of dihydrogen phosphate and sodium hydrogen phosphate); voltage "plus 10 kV"; analysis time passes within 40 minutes; forced air cooling of the capillary to room temperature; output and processing of results on a computer.

The method of capillary electrophoresis to determine the mass concentration of amino acids is based on the separation of anionic forms of N-phenylthiocarbamyl derivatives of amino acids under the influence of an electric field due to their different electrophoretic mobility. To identify and quantify the analyzed components, ultraviolet absorption is recorded at a wavelength of 254 nm.

Alpha amino acids, when reacted with a phenylisothiocyanate in an alkaline medium, produce N-phenylthiocarbamyl (FTK) derivatives, which are acids that exist under basic conditions in the form of anions. Due to the presence in the structure of the obtained compounds of the benzene ring, phenylcarbamyl derivatives have an absorption band at 254 nm. The content of free amino acids is determined through its derivatives with phenylisothiocyanate by capillary electrophoresis.

In the process of analysis, the order and time of release of free amino acids in organs and tissues in clinically healthy animals and for helminthiases was established (Table 1, Table 2, Table 3, Table 4 and Table 5).

Table 1 The order and time of release of free amino acids in an extract in various organs and tissues of clinically healthy pigs No. Component Exit time, min Muscle Heart Liver Lungs Spleen Kidney one Arginine 15,20 16.07 15.25 15.40 16.48 16.15 2 Lysine - 21.15 21.22 - 22.25 20.34 3 Tyrosine - - - - - - four Phenylalanine 20.49 22.41 21.22 24.56 - 5 Histidine 21.52 23.36 22.48 22.35 25.21 - 6 Leucine 26.29 26.23 27.27 26.43 30.15 27.38 7 Methionine 26.34 - 27.34 27.32 31.18 - 8 Valine 27.12 29.01 28.16 28.10 32.05 - 9 Proline 27.32 30.00 28.28 28.14 32.18 - 10 Threonine 28.44 30.50 29.25 29.04 33.56 - eleven Tryptophan 28.57 31.15 29.14 34.05 - 12 Serine 30.00 32.31 30.32 30.08 35.18 - 13 α-Alanine 30.41 33.52 31.36 30.12 36.34 - fourteen Glycine 32.35 35.32 33.28 32.31 39.28 -

table 2 The order and time of release of free amino acids in an extract in various organs and tissues in pigs with metastrongilosis No. Component Exit time, min Muscle Heart Liver Lungs Spleen Kidney one Arginine 14.04 14.37 14.29 14.37 15.37 15,20 2 Lysine - - - 18.26 18,42 20.32 3 Tyrosine 19.06 - - - - 20.39 four Phenylalanine 19.43 19.12 19.32 20.43 20.39 21.34 5 Histidine 20.06 19.46 20.37 21.08 21.21 21.48 6 Leucine 21.19 21.38 23.40 25.02 22.46 26.09 7 Methionine 21.37 22.17 23.57 25.42 22.54 26.55 8 Valine 22.34 22.38 24.36 25.49 23.15 27.25 9 Proline 23.00 23.27 24.43 26.05 23.48 27.49 10 Threonine 23.32 23.57 25.27 26.12 24.53 28.10 eleven Tryptophan 23.51 24.18 25.36 26.35 25.32 - 12 Serine 24.39 24.44 26.49 27.56 26.57 29.13 13 α-Alanine 24.44 25.34 26.54 28.23 27.47 29.26 fourteen Glycine 26.57 26.38 28.39 29.27 28.35 31.41

Table 3 The order and time of release of free amino acids in an extract in various organs and tissues of clinically healthy cattle No. Component Exit time, min Muscle Heart Liver Lungs Spleen Kidney one Arginine 14.22 14.04 - - 14.35 6.43 2 Lysine - - 19.02 - - - 3 Tyrosine - - - 15.48 - - four Phenylalanine - 19.50 19.12 16.51 - 13.24 5 Histidine 20.09 19.27 19.35 20.25 21.39 13.44 6 Leucine 23.12 23.15 22.21 23.35 25.29 17.46 7 Methionine 23.47 24.41 23.52 23,43 25.48 18.23 8 Valine 24.25 24.56 24.01 24.41 26.52 18.52 9 Proline 24.49 25.19 24.40 24.53 27.11 19.37 10 Threonine 25.08 25,50 24.55 - - 19.49 eleven Tryptophan 25.48 - - - - 20.30 12 Serine 26.15 26,42 25.45 26.41 29.02 21.38 13 α-Alanine 26.37 26.49 26.37 26.55 29.56 21.47 fourteen Glycine 28.07 28.25 28.34 28.40 - 23.56

Table 4 The order and time of release of free amino acids in an extract in various organs and tissues in cattle dicroceliosis No. Component Exit time, min Muscle Heart Liver Lungs Spleen Kidney one Arginine 15.48 15.37 15.42 15.10 15.32 15.41 2 Lysine 21.33 21.44 20.46 21.15 21.46 20.37 3 Tyrosine 22.18 - 21.19 22.17 21.57 21.45 four Phenylalanine 22.31 22.24 22.33 22.37 22.55 21.54 5 Histidine 22.44 23.10 22.38 23.21 23.09 22.41 6 Leucine 23.38 27.14 26.54 28.35 27.10 26.40 7 Methionine 27.47 27.37 27.57 29.33 27.23 27.24 8 Valine 28.34 28.39 28.13 30.10 28.46 27.36 9 Proline 28.54 29.25 28.52 31.17 29.14 28.49 10 Threonine 29.20 29.29 29.42 31.82 29.43 29.30 eleven Tryptophan 29.34 30.26 29.48 32.19 30.05 29.37 12 Serine 31.06 31.28 30.57 33.29 31.24 30.45 13 α-Alanine 31.48 32.08 31.54 34.48 31.49 31.33 fourteen Glycine 34.45 34.38 34.40 38.23 34.21 34.27

Table 5 The order and time of release of free amino acids in an extract in various organs and tissues in cattle with echinococcosis No. Component Exit time, min Muscle Heart Liver Lungs Spleen Kidney one Arginine 15.44 15,56 17.45 16.15 16.27 15,53 2 Lysine 17.48 20.43 23.39 19.25 18.36 19.23 3 Tyrosine 19.53 - 26.08 22.21 - - four Phenylalanine 20.05 22.35 26.35 23.37 25.39 02/23 5 Histidine 20.57 26.40 26.53 24.34 26.58 23.57 6 Leucine 24.52 27.10 34.47 29.55 33.11 28.52 7 Methionine 25.48 28.05 35.49 30.21 33.45 29.02 8 Valine 26.36 28.05 37.09 30.27 34.38 29.33 9 Proline 26.39 28.41 38.09 31.13 35.21 29.54 10 Threonine 27.24 28.52 38.53 31.49 36,43 30.51 eleven Tryptophan 27.53 29.42 39.47 - - 31.09 12 Serine 28.47 30,44 40.54 33.26 38.06 32.12 13 α-Alanine 28.53 31.01 41.48 34.21 38.46 32.52 fourteen Glycine 30,42 33.00 45.47 36.37 42,20 34.49

As a result of the studies, it was found that during the invasion of pigs by metastrongylyus, the concentration of free amino acids in the extraction of the longest back muscle was 8 times higher than serine, 6 times higher than proline and threonine, 4 times higher than leucine, 3 times higher than tryptophan, 2 times higher α-alanine, glycine and phenylalanine, 1.2 times methionine, 1.1 times arginine and, conversely, it was 3.5 times lower than histidine, 5 times lower than valine in relation to clinically healthy animals (Table 6 )

During pig invasion by metastrongylyus, the concentration of free amino acids in the extract of the heart muscle was 4.5 times higher than leucine, 2 times higher than histidine, threonine and tryptophan, 1.1 times higher than α-alanine and, conversely, 1.1 times lower arginine and glycine, 1.3 times proline, 2 times serine relative to clinically healthy animals (Table 6).

The concentration of free amino acids during invasion of pigs by metastrongylyus in an extract from liver tissues was 12 times higher than leucine, 4 times higher than histidine, 3 times higher than valine, proline, threonine, tryptophan and phenylalanine, 2 times higher than glycine, methionine and serine, 1.2 times - α-alanine, and, conversely, 1.1 times lower - arginine relative to clinically healthy animals (Table 7).

During pig invasion by metastrongylyus, the concentration of free amino acids in the pulmonary tissue extract was 4 times higher than histidine and phenylalanine, 3 times higher than methionine, valine and proline, 2 times higher than α-alanine, leucine, serine, threonine and tryptophan, 1.2 times - arginine and glycine relative to clinically healthy animals (table 7).

The concentration of free amino acids during invasion of pigs by metastrongylyus in the spleen hood was 2 times higher than leucine, 1.5 times higher than arginine, histidine, methionine, proline, serine and tryptophan, and 1.2 times higher than α-alanine, valine, glycine, lysine , threonine and phenylalanine relative to clinically healthy animals (table 8).

During pig invasion by metastrongylyus, the concentration of free amino acids in the extract of renal tissue was 14 times higher than leucine, 2 times higher than lysine, 1.5 times higher than arginine relative to clinically healthy animals (Table 8).

The total concentration of free amino acids in the extraction of the longest back muscle during pig metastrongilosis was 3316.83 mg / kg of meat, in the heart muscle - 4488.56 mg / kg of meat, in the liver - 7216.61 mg / kg of meat, in the lungs - 5609.94 mg / kg of minced meat, in the spleen - 8269.00 mg / kg of minced meat, in the kidneys - 11439.93 mg / kg of minced meat. The highest content of free amino acids was noted in the renal tissue and was higher than in the extract 3 times that of the longest muscles of the back and heart muscle, 2 times - lung tissue, 1.6 times - the liver and 1.4 times - in the spleen.

Thus, during invasion by the helminths of Metastrongylus elongatus, there was a decrease in the content of bound amino acids and their decomposition into free amino acids and a change in concentration depending on both the functional characteristics of the organ and the location (lungs) of the matured helminths of Metastrongylus elongatus. When pigs were invaded by metastrongyles, it was found that the concentration of the following free amino acids increased 2.3 times in the lung tissue: histidine, phenylalanine, methionine, valine, proline, α-alanine, leucine, serine, threonine, tryptophan, arginine, glycine and lysine relative to clinically healthy animals. In the heart muscle, there was also an increase in the concentration of free amino acids by 1.2 times: leucine, histidine, threonine, tryptophan, and, conversely, a decrease in arginine and glycine, proline, serine relative to clinically healthy animals. A 2-fold increase in the concentration of the following free amino acids took place in the liver: leucine, histidine, valine, proline, threonine, tryptophan, phenylalanine, glycine, methionine, serine, α-alanine, and, conversely, there was lower arginine relative to clinically healthy animals. In the tissues of the spleen, the concentration of the following free amino acids increased 1.4 times: leucine, arginine, histidine, methionine, proline, serine, tryptophan, α-alanine, valine, glycine, lysine, threonine, phenylalanine, relative to clinically healthy animals. In the renal tissue there was an increase in the concentration of free amino acids by 1.4 times: leucine, lysine, arginine relative to clinically healthy animals. The total concentration of free amino acids in the longest muscle of the back, on the contrary, was 1.1 times lower than in clinically healthy animals, however, the tyrosine amino acid decomposed and the concentration of free amino acids increased: serine, proline, threonine, leucine, tryptophan, α-alanine, glycine, phenylalanine, methionine, arginine and, conversely, a decrease in histidine and valine relative to clinically healthy animals.

It should be noted that in clinically healthy animals free amino acids were not detected in the longest muscle of the back and in the lung tissue - lysine and tyrosine; in the heart muscle - tyrosine, methionine and phenylalanine; in the tissues of the liver and spleen - tyrosine, in the renal tissue - tyrosine, phenylalanine, histidine, leucine, methionine, valine, proline, threonine, tryptophan, serine, α-alanine and glycine. This phenomenon, apparently, is associated with the functional features of tissues and organs in the kidney tissue.

During metastrongilosis, the free amino acid lysine was not detected in the longest muscle of the back, which, most likely, underwent decarboxylation, resulting in the release of amines. In the heart muscle and liver tissue during metastrongilosis, free amino acids lysine and tyrosine were not detected, in the lung tissue and spleen tyrosine; tryptophan in the kidney tissue. This phenomenon, most likely, is associated with the functional characteristics of tissues and organs, as well as the influence of metabolic products of Metastrongylus elongatus on the acceleration of the breakdown of protein components. These processes contribute to reducing the quality of meat products.

Table 6 The concentration of free amino acids in the exhaustion of muscle tissue of pigs with metastrongilosis (mg / kg minced meat) Name of amino acids Organs and Tissues Muscle Heart Control Experienced Control Experienced Arginine 196.03 ± 1.43 209.96 ± 1.52 *** 448.65 ± 2.17 427.36 ± 1.52 *** Lysine 0.00 ± 0.00 0.00 ± 0.00 19.69 ± 0.47 0.00 ± 0.00 Tyrosine 0.00 ± 0.00 71.24 ± 0.68 0.00 ± 0.00 0.00 ± 0.00 Phenylalanine 41.73 ± 0.69 80.74 ± 0.60 *** 0.00 ± 0.00 21.63 ± 0.37 Histidine 2445.41 ± 15.50 704.72 ± 2.97 *** 114.63 ± 0.42 167.42 ± 0.76 *** Leucine 16.79 ± 0.33 61.05 ± 0.50 *** 20.36 ± 0.43 91.53 ± 0.59 *** Methionine 140.14 ± 0.54 169.66 ± 0.58 *** 0.00 ± 0.00 459.29 ± 1.19 Valine 137.91 ± 0.95 28.47 ± 0.76 50.59 ± 0.61 62.97 ± 1.05 *** Proline 71.61 ± 0.68 409.16 ± 1.39 *** 894.15 ± 11.89 681.40 ± 6.73 *** Threonine 68.06 ± 0.83 380.76 ± 4.66 *** 191.34 ± 1.13 384.60 ± 2.31 *** Tryptophan 29.20 ± 0.59 88.55 ± 1.06 *** 92.50 ± 0.83 131.80 ± 0.75 *** Serine 7.83 ± 0.09 60.19 ± 0.51 *** 179.15 ± 0.85 91.57 ± 1.51 *** α-Alanine 367.72 ± 0.98 859.70 ± 6.94 *** 1204.78 ± 7.88 1381.35 ± 7.83 *** Glycine 125.45 ± 0.93 192.64 ± 1.10 *** 626.24 ± 1.74 587.64 ± 6.41 *** Total 3647.88 3316.83 3842.08 4488.56 *** P> 0.001

Table 7 The concentration of free amino acids in the pulmonary tissue and liver of pigs in case of metastrongilosis (mg / kg minced meat) Name of amino acids Organs and Tissues Liver Lungs Control Experienced Control Experienced Arginine 159.80 ± 0.76 135.71 ± 0.96 *** 241.06 ± 0.56 287.09 ± 2.40 *** Lysine 54.23 ± 1.29 0.00 ± 0.00 0.00 ± 0.00 30.19 ± 0.82 Tyrosine 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 Phenylalanine 12.86 ± 0.33 39.04 ± 0.53 *** 17.31 ± 0.35 70.61 ± 0.64 *** Histidine 379.56 ± 3.02 1336.86 ± 11.05 *** 267.41 ± 3.32 1058.41 ± 5.84 *** Leucine 40.61 ± 0.51 474.56 ± 3.33 *** 29.45 ± 0.71 59.39 ± 0.99 *** Methionine 341.71 ± 2.49 596.76 ± 3.94 *** 310.74 ± 0.96 1041.50 ± 9.77 *** Valine 119.96 ± 0.44 297.46 ± 5.58 *** 70.80 ± 0.48 220.66 ± 1.97 *** Proline 188.11 ± 1.42 500.56 ± 2.71 *** 139.29 ± 0.79 364.67 ± 3.55 *** Threonine 357.11 ± 3.32 1013.25 ± 4.11 *** 263.94 ± 2.54 540.81 ± 3.10 *** Tryptophan 249.39 ± 3.11 520.61 ± 5.26 *** 130.41 ± 0.70 197.53 ± 1.45 *** Serine 124.04 ± 1.02 298.78 ± 1.49 *** 79.00 ± 0.69 161.91 ± 0.92 *** α-Alanine 1413.58 ± 15.01 1589.51 ± 10.24 *** 471.72 ± 4.02 1017.23 ± 5.61 *** Glycine 178.09 ± 0.81 413.51 ± 3.58 *** 465.98 ± 5.74 559.94 ± 13.23 *** Total 3619.05 7216.61 2487.11 5609.94 *** P> 0.001

Table 8 The concentration of free amino acids in the exhaust tissue of the spleen and kidneys of pigs with metastrongilia (mg / kg minced meat) Name of amino acids Organs and Tissues Spleen Kidney Control Experienced Control Experienced Arginine 606.75 ± 6.01 883.42 ± 22.16 *** 284.78 ± 2.10 428.75 ± 1.99 *** Lysine 65.83 ± 1.03 82.05 ± 0.71 *** 9.73 ± 0.21 17.68 ± 0.32 *** Tyrosine 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 168.45 ± 0.87 Phenylalanine 93.08 ± 0.68 105.57 ± 1.99 *** 0.00 ± 0.00 82.00 ± 0.59 Histidine 549.79 ± 4.33 847.07 ± 11.65 *** 0.00 ± 0.00 2126.67 ± 9.68 Leucine 306.59 ± 2.73 636.49 ± 8.18 *** 12.57 ± 0.23 170.93 ± 0.62 *** Methionine 467.13 ± 2.55 755.80 ± 12.65 *** 0.00 ± 0.00 2803.88 ± 11.51 Valine 368.60 ± 1.50 477.45 ± 4.15 *** 0.00 ± 0.00 503.96 ± 4.01 Proline 363.70 ± 2.97 498.91 ± 3.38 *** 0.00 ± 0.00 792.64 ± 4.01 Threonine 647.33 ± 1.76 706.05 ± 3.73 *** 0.00 ± 0.00 716.88 ± 5.93 Tryptophan 467.60 ± 2.78 651.80 ± 7.68 *** 0.00 ± 0.00 0.00 ± 0.00 Serine 243.16 ± 1.22 386.95 ± 5.72 *** 0.00 ± 0.00 495.59 ± 3.25 α-Alanine 1105.23 ± 12.00 1380.53 ± 13.52 *** 0.00 ± 0.00 2653.08 ± 14.46 Glycine 714.78 ± 10.78 856.91 ± 7.97 *** 0.00 ± 0.00 479.42 ± 4.31 Total 5999.57 8269.00 307.08 11,439.93 *** P> 0.001

As a result of the studies, it was found that during invasion of cattle Echinococcus granulosus, the concentration of free amino acids in the extract of the longest muscle of the back was 77 times higher than histidine, 2 times higher than glycine, methionine and proline, 1.3 times higher than α-alanine, arginine and serine, 1.2 times - valine, 1.1 times - leucine and threonine relative to clinically healthy animals. Free amino acids tryptophan (70.65 ± 0.052 mg / kg) and phenylalanine (76.48 ± 0.63 mg / kg) were recorded in cattle with echinococcosis (Table 9).

During invasion of cattle Echinococcus granulosus, the concentration of free amino acids in the extract of the heart muscle was 57 times higher than methionine, 8 times higher than glycine, 3 times higher than serine, 5 times higher than arginine and phenylalanine, 2 times higher than α-alanine, valine, histidine, proline and threonine, 1.3 times - leucine relative to clinically healthy animals. The free amino acid tyrosine was not detected in both clinically healthy animals and echinococcosis. Free amino acids lysine (16.94 ± 0.48 mg / kg) and tryptophan (141.90 ± 1.04 mg / kg) were recorded in cattle with echinococcosis (Table 9).

The concentration of free amino acids during invasion of cattle Echinococcus granulosus in a liver extract was 6 times higher than lysine, 5 times higher than histidine and phenylalanine, 3 times higher than methionine, 2 times higher than glycine and leucine, 1.5 times higher than threonine, 1.3 times - serine, 1.2 times - proline, 1.1 times - α-alanine relative to clinically healthy animals. The concentration of free amino acid valine was almost at the level of clinically healthy animals. Free amino acids arginine (324.22 ± 0.90 mg / kg), lysine (85.80 ± 0.68 mg / kg) were recorded in cattle with echinococcosis (Table 10). During invasion of cattle Echinococcus granulosus, the concentration of free amino acids in the pulmonary tissue extract was 9 times higher than methionine, 5 times higher than histidine and phenylalanine, 4 times higher than proline, 3 times higher than α-alanine, 2 times higher than glycine and leucine , 1.4 times - serine, 1.3 times - valine relative to clinically healthy animals. Free amino acids arginine (252.48 ± 1.06 mg / kg), lysine (12.56 ± 0.50 mg / kg) and threonine (177.14 ± 2.16 mg / kg) were recorded in cattle with echinococcosis. The free amino acid tryptophan was not detected in both clinically healthy animals and echinococcosis (Table 10).

The concentration of free amino acids during invasion of cattle Echinococcus granulosus in the extract of the spleen was 16 times higher than methionine, 13 times higher than serine, 11 times higher than histidine, 10 times higher than valine, 8 times higher than arginine, 6 times higher than α- alanine, 4 times - proline, 3 times - leucine relative to clinically healthy animals. Free amino acids glycine (7.35.84 ± 1.51 mg / kg), lysine (19.30 ± 0.30 mg / kg) and threonine (919.00 ± 1.66 mg / kg) were recorded in cattle livestock with echinococcosis. The free amino acids tyrosine and tryptophan were not detected both in clinically healthy animals and in echinococcosis (Table 11).

During the invasion of cattle by Echinococcus granulosus, the concentration of free amino acids in the extract of the renal tissue was 3 times higher than phenylalanine, 2 times higher than arginine, glycine and methionine, 1.2 times higher than threonine and, conversely, 3 times lower than leucine, 2 times lower times - valine and tryptophan relative to clinically healthy animals. The concentration of free amino acids of α-alanine, proline and serine was almost at the level with clinically healthy animals. The free amino acid lysine (8.23 ± 0.32 mg / kg) was recorded in cattle with echinococcosis. The free amino acid tyrosine was not detected in both clinically healthy animals and echinococcosis (Table 11).

The total concentration of free amino acids in the extract of the longest muscle of the back with echinococcosis was 32,403.95 mg / kg of minced meat, in the heart muscle - 6,718.14 mg / kg of minced meat, in the liver - 11,422.98 mg / kg of minced meat, in the lungs - 4,094.93 mg / kg of minced meat, in the spleen - 9123.24 mg / kg of minced meat, in the kidneys - 3341.76 mg / kg of minced meat. The highest content of free amino acids was noted in the stretching of the longest back muscle and was 9 times higher than in the stretching of the kidneys; heart muscle - 5 times; liver - 3 times, lungs - 8 times, spleen - 4 times.

Thus, during the invasion of cattle by Echinococcus granulosus, there was a 2-fold increase in the concentration of free amino acids in the liver tissues, by a factor of 15 in the longest muscle of the back, by a factor of 3.4 in the heart muscle and lung tissue, and by an increase of 11 in the spleen tissue times in the kidney tissue - 1.1 times relative to clinically healthy animals. During invasion of cattle Echinococcus granulosus, the free amino acid tyrosine was not detected in the heart muscle, tryptophan in the lung tissue, tyrosine and tryptophan in the spleen tissue.

It should be noted that clinically healthy animals did not reveal free amino acids: in the longest muscle of the back - lysine, tryptophan, tyrosine and phenylalanine, in the heart muscle - lysine, tryptophan and tyrosine, in the liver tissues - arginine, tryptophan and tyrosine, in the lung tissue - arginine, lysine, threonine and tryptophan, in the tissues of the spleen - lysine, tryptophan, tyrosine and phenylalanine, in the renal tissue - tyrosine. This phenomenon, apparently, is associated with the functional features of tissues and organs.

Table 9 The concentration of free amino acids in the extract of muscle tissue in cattle with echinococcosis (mg / kg minced meat) Name of amino acids Organs and Tissues Muscle Heart Control Experienced Control Experienced Arginine 344.48 ± 0.86 455.92 ± 2.66 *** 360.52 ± 2.01 564.80 ± 1.83 *** Lysine 0.00 ± 0.00 830.75 ± 0.93 0.00 ± 0.00 16.94 ± 0.48 Tyrosine 0.00 ± 0.00 36.01 ± 0.52 0.00 ± 0.00 0.00 ± 0.00 Phenylalanine 0.00 ± 0.00 70.65 ± 0.052 9.46 ± 0.23 46.62 ± 0.51 *** Histidine 376.68 ± 1.95 29067.34 ± 264.16 *** 74.39 ± 0.39 126.21 ± 0.62 *** Leucine 64.56 ± 0.29 71.22 ± 0.61 * 61.06 ± 0.44 77.80 ± 0.72 *** Methionine 186.69 ± 0.51 299.64 ± 1.99 *** 513.81 ± 0.62 2947.93 ± 7.45 *** Valine 350.47 ± 0.49 435.03 ± 3.22 *** 36.46 ± 0.32 874.18 ± 4.47 *** Proline 73.80 ± 0.43 40.63 ± 0.51 *** 166.45 ± 0.66 95.86 ± 0.39 *** Threonine 114.03 ± 0.29 102.99 ± 1.38 ** 66.67 ± 0.48 131.95 ± 0.96 *** Tryptophan 0.00 ± 0.00 76.48 ± 0.63 0.00 ± 0.00 141.90 ± 1.04 Serine 55.79 ± 0.57 71.64 ± 0.63 *** 42.28 ± 0.72 130.48 ± 0.80 *** α-Alanine 500.15 ± 1.43 668.82 ± 1.25 *** 633.30 ± 1.18 1232.98 ± 0.99 *** Glycine 106.60 ± 0.82 176.83 ± 0.59 *** 40.15 ± 0.48 330.49 ± 0.73 *** Total 2173.25 32,403.95 2004.55 6718.14 * P <0.05; ** P> 0.01; *** P> 0.001

Table 10 The concentration of free amino acids in the pulmonary tissue and liver of cattle in echinococcosis (mg / kg of minced meat) Name of amino acids Organs and Tissues Liver Lungs Control Experienced Control Experienced Arginine 0.00 ± 0.00 324.22 ± 0.90 0.00 ± 0.00 252.48 ± 1.06 Lysine 15.27 ± 0.52 85.80 ± 0.68 0.00 ± 0.00 12.56 ± 0.50 Tyrosine 0.00 ± 0.00 579.47 ± 2.83 48.20 ± 0.64 53.38 ± 0.55 ** Phenylalanine 7.79 ± 0.13 35.30 ± 0.50 *** 13.12 ± 0.27 60.15 ± 0.51 *** Histidine 215.06 ± 0.38 1043.66 ± 3.73 *** 136.88 ± 054 669.34 ± 4.84 *** Leucine 176.68 ± 0.31 412.58 ± 2.51 *** 41.71 ± 0.75 71.81 ± 0.85 *** Methionine 561.55 ± 2.21 1711.92 ± 7.11 *** 64.24 ± 0.60 596.86 ± 2.84 *** Valine 440.92 ± 0.43 464.06 ± 2.55 ** 96.12 ± 0.47 122.05 ± 1.35 *** Proline 443.06 ± 0.71 563.11 ± 8.90 *** 71.19 ± 0.51 287.43 ± 5.41 *** Threonine 981.95 ± 0.64 1497.78 ± 2.19 *** 0.00 ± 0.00 177.14 ± 2.16 Tryptophan 0.00 ± 0.00 538.36 ± 1.26 0.00 ± 0.00 0.00 ± 0.00 Serine 381.52 ± 0.60 503.68 ± 2.63 *** 22.20 ± 0.43 31.33 ± 0.71 * α-Alanine 2110.61 ± 12.30 2362.55 ± 2.05 *** 326.54 ± 5.08 899.37 ± 3.10 *** Glycine 597.79 ± 1.49 1300.49 ± 2.81 *** 368.18 ± 0.77 861.03 ± 1.62 *** Total 5932,20 11422.98 1188.38 4094.93 * P <0.05; ** P> 0.01; *** P> 0.001

Table 11 The concentration of free amino acids in the exhaust tissue of the spleen and kidneys of cattle with echinococcosis (mg / kg minced meat) Name of amino acids Organs and Tissues Spleen Kidney Control Experienced Control Experienced Arginine 49.95 ± 0.74 380.30 ± 0.51 *** 75.67 ± 0.64 160.10 ± 0.65 *** Lysine 0.00 ± 0.00 19.30 ± 0.30 0.00 ± 0.00 8.23 ± 0.32 Tyrosine 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 Phenylalanine 0.00 ± 0.00 191.73 ± 1.38 29.64 ± 0.34 74.64 ± 0.85 *** Histidine 160.94 ± 0.37 1806.66 ± 4.45 *** 436.87 ± 0.48 441.53 ± 0.86 Leucine 50.49 ± 0.47 171.42 ± 1.39 *** 157.46 ± 0.55 46.07 ± 0.77 *** Methionine 87.71 ± 0.70 1454.22 ± 8.98 *** 207.67 ± 0.59 401.63 ± 3.53 *** Valine 51.92 ± 0.47 532.82 ± 7.51 *** 142.31 ± 0.66 61.96 ± 0.78 *** Proline 79.93 ± 0.44 305.90 ± 5.06 *** 224.69 ± 0.59 230.26 ± 1.69 ** Threonine 0.00 ± 0.00 919.00 ± 1.66 473.16 ± 1.61 595.07 ± 1.74 *** Tryptophan 0.00 ± 0.00 0.00 ± 0.00 164.37 ± 0.71 79.15 ± 0.48 *** Serine 26.45 ± 0.48 331.21 ± 0.87 *** 130.09 ± 0.72 131.08 ± 0.79 α-Alanine 359.93 ± 0.54 2274.84 ± 2.51 *** 798.67 ± 2.14 830.10 ± 0.62 *** Glycine 0.00 ± 0.00 735.84 ± 1.51 373.36 ± 0.66 581.94 ± 0.78 *** Total 867.32 9123.24 3213.96 3641.76 * P <0.05; ** P> 0.01; *** P> 0.001

As a result of the studies, it was found that during the invasion of Dicrocoelium lanceatum cattle, the concentration of free amino acids in the extract of the longest muscle of the back was 4 times higher than methionine, 3 times higher than valine, 2 times higher than α-alanine, leucine, proline, threonine and serine , 1.6 times - glycine, 1.3 times - arginine relative to clinically healthy animals. The concentration of histidine free amino acid was almost at the level of clinically healthy animals. Free amino acids tryptophan (161.65 ± 1.48 mg / kg) and phenylalanine (38.96 ± 0.66 mg / kg) were recorded in cattle with dicroceliosis (Table 12).

During the invasion of Dicrocoelium lanceatum cattle, the concentration of free amino acids in the extract of the heart muscle was 19 times higher than valine, 7 times higher than glycine and methionine, 4 times higher than histidine and threonine, 3 times higher than α-alanine, serine and phenylalanine, 2 times times - leucine, 1.4 times - proline, 1.2 times - arginine relative to clinically healthy animals. The free amino acid tyrosine was not detected in both clinically healthy animals and dicroceliosis. Free amino acids lysine (92.12 ± 1.26 mg / kg) and tryptophan (151.57 ± 1.50 mg / kg) were recorded in cattle with dicroceliosis (Table 12).

The concentration of free amino acids during the invasion of cattle Dicrocoelium lanceatum in a liver extract was 45 times higher than phenylalanine, 15 times higher than histidine, 7 times higher than leucine, 3 times higher than glycine, lysine and proline, 2 times higher than valine, methionine and threonine, 1.5 times - α-alanine and serine relative to clinically healthy animals. Free amino acids arginine (274.49 ± 4.39 mg / kg), tyrosine (237.93 ± 1.01 mg / kg) and tryptophan (898.62 ± 3.22 mg / kg) were recorded in cattle with dicroceliosis (Table 13).

During the invasion of Dicrocoelium lanceatum cattle, the concentration of free amino acids in the pulmonary tissue extract was 76 times higher than leucine, 70 times higher than serine, 54 times higher than methionine, 50 times higher than histidine, 44 times higher than proline, 32 times higher than phenylalanine , 30 times - valine, 26 times - α-alanine, 10 times - glycine, 1.7 times - tyrosine relative to clinically healthy animals. The free amino acids arginine (1377.42 ± 5.69 mg / kg), lysine (127.45 ± 1.19 mg / kg), threonine (4272.39 ± 6.08 mg / kg) and tryptophan (3582.97 ± 6.65 mg / kg) have been reported in cattle with dicroceliosis (Table 13).

The concentration of free amino acids during invasion of cattle Dicrocoelium lanceatum in the spleen hood was 34 times higher than leucine, 29 times higher than serine, 28 times higher than valine, 26 times higher than histidine, 25 times higher than methionine, 20 times higher than proline, 14 times - arginine, 11 times - α-alanine relative to clinically healthy animals. Free amino acids glycine (1609.90 ± 6.64 mg / kg), lysine (48.69 ± 0.78 mg / kg), threonine (2254.49 ± 8.60 mg / kg), tyrosine (50.84 ± 0.49 mg / kg) and tryptophan (1868.65 ± 7.66 mg / kg) were recorded in cattle with dicroceliosis (Table 14).

During the invasion of Dicrocoelium lanceatum cattle, the concentration of free amino acids in the extract of the renal tissue was 8 times higher than histidine, 7 times higher than arginine, 6 times higher than valine, 4 times higher than leucine, methionine and phenylalanine, 3 times lower than proline and tryptophan , 2 times - serine and threonine, 1.4 times - glycine, 1.3 times - α-alanine relative to clinically healthy animals. The free amino acid lysine (60.36 ± 1.16 mg / kg) was recorded in cattle with dicroceliosis (Table 14).

The total concentration of free amino acids in the extract of the longest muscle of the back with dicroceliosis was 5414.47 mg / kg of meat, in the heart muscle - 7884.32 mg / kg of meat, in the liver - 17281.74 mg / kg of meat, in the lungs - 3013.85 mg / kg of minced meat, in the spleen - 22,400.37 mg / kg of minced meat, in the kidneys - 0348.98 mg / kg of minced meat. The highest content of free amino acids was noted in the pulmonary tissue extract and was 8 times higher than in the extract of the longest back muscle; heart muscle - 5 times; liver - 3 times, spleen - 2 times, kidneys - 4 times.

Thus, during the invasion of Dicrocoelium lanceatum cattle, there was an increase in the total concentration of free amino acids in the stretching of the longest muscle of the back by 2.5 times, of the heart muscle - by 4 times, of the liver and kidneys - by 3 times, of the lungs - by 36 times, of the spleen - 26 times relatively clinically healthy animals.

During cattle invasion by dicrocelia, the tyrosine free amino acid was not detected in the heart muscle.

Table 12 The concentration of free amino acids in the muscle tissue of cattle during dicroceliosis (mg / kg minced meat) Name of amino acids Organs and Tissues Muscle Heart Control Experienced Control Experienced Arginine 344.48 ± 0.86 459.62 ± 5.24 ** 360.52 ± 2.01 439.44 ± 11.27 ** Lysine 0.00 ± 0.00 428.40 ± 3.64 0.00 ± 0.00 92.12 ± 1.26 Tyrosine 0.00 ± 0.00 134.02 ± 1.08 0.00 ± 0.00 0.00 ± 0.00 Phenylalanine 0.00 ± 0.00 38.96 ± 0.66 9.46 ± 0.23 29.71 ± 0.71 *** Histidine 376.68 ± 1.95 378.26 ± 5.49 *** 74.39 ± 0.39 *** 292.91 ± 3.46 *** Leucine 64.56 ± 0.29 130.83 ± 1.20 *** 61.06 ± 0.44 119.90 ± 1.01 *** Methionine 186.69 ± 0.51 704.37 ± 4.24 *** 513.81 ± 0.62 3470.57 ± 6.97 *** Valine 350.47 ± 0.49 937.55 ± 7.55 *** 36.46 ± 0.32 681.31 ± 7.68 *** Proline 73.80 ± 0.43 132.18 ± 1.63 *** 166.45 ± 0.66 120.02 ± 1.36 *** Threonine 114.03 ± 0.29 255.98 ± 8.79 *** 66.67 ± 0.48 279.92 ± 3.19 *** Tryptophan 0.00 ± 0.00 161.65 ± 1.48 *** 0.00 ± 0.00 151.57 ± 1.50 Serine 55.79 ± 0.57 102.05 ± 1.51 *** 42.28 ± 0.72 118.15 ± 0.89 *** α-Alanine 500.15 ± 1.43 984.64 ± 3.54 *** 633.30 ± 1.18 1817.13 ± 6.36 *** Glycine 106.60 ± 0.82 165.96 ± 1.12 *** 40.15 ± 0.48 271.57 ± 1.56 *** Total 2173.25 5414.47 2004.55 7884.32 ** P> 0.01; *** P> 0.001

Table 13 The concentration of free amino acids in the pulmonary tissue and liver of cattle under dicroceliosis (mg / kg minced meat) Name of amino acids Organs and Tissues Liver Lungs Control Experienced Control Experienced Arginine 0.00 ± 0.00 274.49 ± 4.39 0.00 ± 0.00 1377.42 ± 5.69 Lysine 15.27 ± 0.52 50.93 ± 0.64 *** 0.00 ± 0.00 127.45 ± 1.19 Tyrosine 0.00 ± 0.00 237.93 ± 1.01 48.20 ± 0.64 80.56 ± 0.56 *** Phenylalanine 7.79 ± 0.13 354.00 ± 2.84 **** 13.12 ± 0.27 415.93 ± 28.63 *** Histidine 215.06 ± 0.38 3127.78 ± 8.59 *** 136.88 ± 054 6802.74 ± 5.27 *** Leucine 176.68 ± 0.31 1156.42 ± 7.16 *** 41.71 ± 0.75 3155.81 ± 10.70 *** Methionine 561.55 ± 2.21 1162.26 ± 8.89 *** 64.24 ± 0.60 3494.40 ± 4.03 *** Valine 440.92 ± 0.43 860.74 ± 7.77 *** 96.12 ± 0.47 2840.98 ± 7.66 *** Proline 443.06 ± 0.71 1092.69 ± 4.89 *** 71.19 ± 0.51 3157.80 ± 8.10 *** Threonine 981.95 ± 0.64 2368.00 ± 7.71 *** 0.00 ± 0.00 4272.39 ± 6.08 *** Tryptophan 0.00 ± 0.00 898.62 ± 3.22 0.00 ± 0.00 3582.97 ± 6.65 *** Serine 381.52 ± 0.60 594.53 ± 39.67 *** 22.20 ± 0.43 1561.15 ± 10.92 *** α-Alanine 2110.61 ± 12.30 3152.38 ± 9.11 *** 326.54 ± 5.08 8482.59 ± 8.54 *** Glycine 597.79 ± 1.49 1950.97 ± 5.91 *** 368.18 ± 0.77 ± 3661.66 ± 8.50 *** Total 5932,20 17,281.74 1188.38 43,013.85 *** P> 0.001

Table 14 The concentration of free amino acids in the exhaust tissue of the spleen and kidneys of cattle with dicroceliosis (mg / kg minced meat) Name of amino acids Organs and Tissues Spleen Kidney Control Experienced Control Experienced Arginine 49.95 ± 0.74 707.05 ± 5.24 *** 75.67 ± 0.64 524.14 ± 6.71 *** Lysine 0.00 ± 0.00 48.69 ± 0.78 0.00 ± 0.00 60.36 ± 1.16 Tyrosine 0.00 ± 0.00 50.84 ± 0.49 0.00 ± 0.00 57.62 ± 0.78 Phenylalanine 0.00 ± 0.00 150.72 ± 0.66 29.64 ± 0.34 127.90 ± 1.03 *** Histidine 160.94 ± 0.37 4119.35 ± 8.63 *** 436.87 ± 0.48 3523.08 ± 10.84 *** Leucine 50.49 ± 0.47 1692.98 ± 4.25 *** 157.46 ± 0.55 614.80 ± 5.13 *** Methionine 87.71 ± 0.70 2172.77 ± 6.15 *** 207.67 ± 0.59 809.93 ± 6.55 *** Valine 51.92 ± 0.47 1466.10 ± 7.17 *** 142.31 ± 0.66 883.83 ± 7.82 *** Proline 79.93 ± 0.44 1592.88 ± 3.11 *** 224.69 ± 0.59 569.57 ± 7.23 *** Threonine 0.00 ± 0.00 2254.49 ± 8.60 473.16 ± 1.61 893.58 ± 4.43 *** Tryptophan 0.00 ± 0.00 1868.65 ± 7.66 164.37 ± 0.71 463.91 ± 4.03 *** Serine 26.45 ± 0.48 779.05 ± 4.39 *** 130.09 ± 0.72 267.25 ± 5.87 *** α-Alanine 359.93 ± 0.54 3886.90 ± 7.64 *** 798.67 ± 2.14 1020.99 ± 4.08 *** Glycine 0.00 ± 0.00 1609.90 ± 6.64 373.36 ± 0.66 532.02 ± 7.61 *** Total 867.32 22,400.37 3213.96 10348.98 *** P> 0.001

By the concentration, order and time of release of free amino acids, it is possible to establish the quality of animal slaughter products during helminthiases. The identification of the concentration of free amino acids in the extract of muscle tissue and organs is important for establishing the quality and safety of the slaughter products of clinically healthy pigs, as well as for metastrongylosis; dicroceliosis and echinococcosis of cattle. A high concentration of free amino acids in animal helminthiases indicates the processes of protein breakdown in tissues and organs. With helminthiases, a change in the concentration of free amino acids occurs, depending on both the functional characteristics of the organ and the location of the helminths. Consequently, according to the concentration of free amino acids, it can be established that for metastrongilosis of pigs, dicroceliosis and echinococcosis of cattle, internal organs should be sent for technical utilization, carcasses for industrial processing (production of cooked and boiled-smoked sausages) in connection with destructive processes in the tissues of slaughter animals. In order to ensure the quality and safety of animal slaughter products during metastrongylosis, dicroceliosis and echinococcosis during primary processing in production and sales, it is necessary to: increase control of technological production, the level of laboratory analysis of animal slaughter products, including determination of free amino acid concentration using capillary electrophoresis “Drops 103- P "to establish the degree of tissue destruction caused by the waste products of helminths. The free amino acids were subsequently subjected to decarboxylation, resulting in the release of ammonium. With an increase in invasion, a slight decrease in the concentration of ammonium was observed, however, it exceeded the maximum permissible norms. This circumstance, most likely, is associated with further decomposition of ammonium into less toxic or non-toxic products of protein breakdown. During deamination of free amino acids, volatile fatty acids were formed (butyric, isobutyric, etc.), and during decarboxylation, ammonium, histamine, and methane. Substances formed during decarboxylation are toxic to the human body.

Claims (1)

A method for assessing the quality of animal slaughter products, including derivatization of a bioassay based on a solution of phenylisothiocyanate in isopropyl alcohol, exposure to it by capillary zone electrophoresis using a working buffer based on β-cyclodextrin, detection of free amino acids and their qualitative and quantitative analysis, characterized in that as bioassays using aqueous extract of animal organs and tissues, in selected bioassays pre microvessel of not more than 0.05 cm ³ of liquid, which is added to 0.1 with ³ 0.1 M sodium carbonate solution, stirred, and then added at least 0.3 cm ³ phenyl isothiocyanate solution in isopropyl alcohol and left to react to 40 minutes, after the contents dried in vivo, is not added more than 0.5 cm ^{3 of} distilled water, stirred, poured into an Eppendorf tube and centrifuged to remove gases and suspensions, then the obtained data as a result of qualitative and quantitative analyzes are compared with the control result of the concentration of free amino acids clinically of pre-domestic animals and determine the quality of products, if the concentration of free amino acids is increased compared with the control, then the products of slaughter of animals are considered infected helminths.

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