RU2613470C2 - Method of reduction of heavy metal content of cow's milk - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: invention relates to animal husbandry, in particular to a method of reduction of the heavy metal content in cow's milk. The method includes the use of a chitosan solution and probiotic "EM-Vita". The feed is given to animals, which contains 2.7-3.3% solution of water-soluble chitosan with the molecular weight of 38 kDa and the deacetylation level of 85% at the rate of 1.8-2.2 ml/kg body weight of the animal, and the probiotic "EM-Vita" at the rate of 18-22 ml per animal. The agents are used once a day during three 9-11-day courses with an interval of 6-8 days.

EFFECT: use of the invention enables to reduce the heavy metal content in the cow's milk and to improve the quality characteristics of milk.

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Description

The invention relates to animal husbandry and can be used to reduce the content of heavy metals in cow's milk.

There are a large number of ways to reduce the content of heavy metals in cow’s milk, involving the use of various means to reduce the level of heavy metals in the body and milk of cows: bio vermiculite, zeolite of the Kamyshlovsky field, shatrashanit and others [1, 2, 3].

There is a method of reducing the content of heavy metals in cow's milk by introducing biovermiculite (expanded vermiculite), which is a natural mineral from the hydromica group, into the diet in the amount of 100 g per head per day [1]. However, the disadvantage of using biovermiculite is the simultaneous absorption of essential trace elements involved in the regulation of body functions with heavy metals.

There is a method of reducing the content of heavy metals in cow's milk by adding to the main diet of the zeolite of the Kamyshlovsky deposit of the Sverdlovsk region at the rate of 0.15-0.20 g / kg of animal body weight once for 25-30 days. Natural zeolites are microporous crystalline aluminosilicates of a crystalline structure containing channels and voids occupied by large ions and water molecules. They have significant freedom of movement, which leads to ion exchange and reversible dehydration. Due to the strictly defined pore sizes of internal cavities, natural zeolites have molecular sieve properties and are good adsorbents for many inorganic and organic substances, including salts of heavy metals [2]. The disadvantage of this method is its limited ability - zeolite Kamyshlovsky deposits, as enterosorbent allows to reduce only the level of Nickel and lead in the body of cows.

A known method of reducing the content of heavy metals in cow's milk by feeding animals shatrashanit (zeolite-containing raw materials produced in the village of Shatrashany, Drozhzhanovsky district of the Republic of Tatarstan). The mineral is fed to cows in dry form in a mixture with concentrated feed at the rate of 300 g per head per day or 2.5-3.0% of the dry matter of the diet [3]. The disadvantage of this method: the lack of selectivity with respect to chemical elements.

The closest analogue is a method of reducing the content of heavy metals in cow's milk by introducing into the diet chitosan acid-soluble with a molecular weight of 250 kDa and a degree of deacetylation of 88% based on 60 mg / kg of animal body weight 2 times a day for 5 days. Re-use is carried out after 5 days [4]. The disadvantage of this method is that acid-soluble chitosan in the gastrointestinal tract manifests itself as a universal sorbent capable of binding a huge range of substances of an organic and inorganic nature, which, ultimately, can adversely affect the scar microbiocenosis. In addition, the adsorption, chelating and complexing properties of acid-soluble chitosan are lower compared to the water-soluble form of chitosan, the effectiveness of which is due to the ability to dissolve in water and saliva.

The aim of the invention was to develop an effective way to reduce the content of heavy metals in milk of cows. This goal is achieved by the fact that cows kept under conditions of technogenic agroecosystems are given with food a 2.7-3.3% solution of chitosan water-soluble with a molecular weight of 38 kDa and a degree of deacetylation of 85% from 1.8-2.2 ml / kg body weight of the animal and the probiotic "EM-Vita" at the rate of 18-22 ml per animal, the drugs are used once a day for three 9-11-day courses with an interval of 6-8 days.

A comparative analysis of the claimed solution with the prototype shows that the method differs from the known one in that they use a 2.7-3.3% solution of chitosan water-soluble with a molecular weight of 38 kDa and a degree of deacetylation of 85% based on 1.8-2.2 ml / kg of animal body weight once a day for three 9-11-day courses with an interval of 6-8 days, which when ingested in the gastrointestinal tract is split, absorbed into the blood and absorbed by the body in the form of low molecular weight compounds - aminoglucans, which stimulate natural resist ntnost body. In addition to chitosan, the probiotic EM-Vita is prescribed at the rate of 18-22 ml per animal according to the above scheme.

The mechanism of action of the EM-Vita probiotic is that when it is used, the number of beneficial bacteria in the gastrointestinal tract increases, which have a depressing effect on pathogenic and conditionally pathogenic microorganisms, improve the population composition of indigenous microflora, contributing to the creation of a favorable environment for metabolic processes in the intestines.

Thus, the claimed method meets the condition of patentability "novelty." Comparison of the proposed method not only with the prototype, but also with other solutions in this field allowed us to identify signs that distinguish the claimed solution from the known, which makes it possible to conclude that it meets the criterion of "significant differences".

Analysis of the differences of the claimed technical solution and known solutions from the prior art allows us to conclude that the proposed method meets the condition of patentability "inventive step".

A method of reducing the content of heavy metals in cow's milk meets the condition of patentability of the invention “industrial applicability”, since the proposed method can be repeatedly used in agricultural enterprises of the agro-industrial complex, regardless of their form of ownership under conditions of technogenic agroecosystems. This is proved by the results of studies showing that water-soluble chitosan in combination with the EM-Vita probiotic are biocompatible with the natural environment, capable of biodegradation to natural products of the biotype; in conditions of technogenic agroecosystems (complex pollution of the animal habitat with salts of heavy metals), the use of water-soluble chitosan in combination with a probiotic helps to eliminate heavy metals from the body of cows; qualitative characteristics of cow's milk meet the requirements of SanPiN.

Water-soluble chitosan is used in combination with the EM-Vita probiotic obtained under the production conditions of Bioprogress CJSC VNIIiTIBP in the city of Schelkovo, Moscow Region. "EM-Vita" consists of a mixture of strains of Laktobacillus plantarum 376B, Lactobacillus casei MDP-1, Saccharomyces cerevisiae, molasses and water. 1 cm ³ contains at least 1 × 10 ⁷ CFU (colony forming units) of lactic acid bacteria and 1 × 10 ⁴ CFU of yeast. Cows kept under conditions of technogenic agroecosystems are given with food a 2.7-3.3% solution of chitosan with a molecular weight of 38 kDa and a degree of deacetylation of 85% based on 1.8-2.2 ml / kg of animal body weight and probiotic "EM-Vita" at the rate of 18-22 ml per animal, drugs are used once a day for three 9-11-day courses with an interval of 6-8 days.

The probiotic “EM-Vita” normalizes the microflora of animal rumen, helping to improve the digestibility of feed, improves immunity, reproductive ability and quality of the products obtained, improves the appearance and health of animals. The microorganisms included in the composition have an antagonistic effect against the simplest pathogenic microorganisms and putrefactive microflora.

Chitosan is a biopolymer of a polysaccharide nature, isolated from shellfish. The main characteristics of chitosan are the degree of acetylation, molecular weight and physical condition. The water-soluble low molecular weight form of chitosan has an effect on a number of non-specific processes characteristic of the body's response to any adverse effect.

Chitosan has adsorption, film-forming, anti-inflammatory, bacteriostatic and antacid properties. Obviously, these functions are performed by chitosan due to the rapidly assimilable fraction in oral administration - a mixture of monomer and its dimer. In addition, water-soluble chitosan has a wide spectrum of antimicrobial activity, including in relation to pathogenic microflora, and is able to stimulate the growth of bifidobacteria and lactobacilli, which allows its use in combination with probiotic preparations.

It is important to note that the adsorption, chelating, and complexing properties of the water-soluble low molecular weight form of chitosan are significantly enhanced in comparison with the use of the acid-soluble form of chitosan.

Biocompatibility, bioavailability, non-toxicity, selectivity with respect to chemical elements and the wide spectrum of action of chitosan determine its compliance with modern requirements and trends in the production of environmentally friendly products. The development and implementation of such products in dairy farming is relevant not only to improve the qualitative and quantitative characteristics of the final product - milk, but also to maintain the optimal level of animal health indicators. The inclusion of such drugs in the program at various stages of implementation can significantly increase its effectiveness.

All of the above served as the basis for studying the effect of water-soluble chitosan with a molecular weight of 38 kDa and a degree of deacetylation of 85% in combination with the probiotic "EM-Vit" on the reduction of heavy metals in cow's milk.

Execution example. To implement the proposed method, an experiment was conducted on the basis of Zaozerny LLC Varna district of Chelyabinsk region. The experiment was conducted on cows of black-motley breed, selected according to the principle of analogues, with a live weight of 450-500 kg, 3 lactations. According to the principle of analogues, cows were divided into 3 groups of 30 animals each. Feeding and caring for animals were similar.

The quality of livestock production largely depends on the characteristics of the biotic cycle in the agroecosphere and the geochemistry of agricultural landscapes. The geochemical situation formed in agrarian landscapes affects the conditions of mineral nutrition of plants, and through trophic chains it affects the state of animals and humans. In this regard, the degree of contamination of the links of the trophic chain by heavy metals of Zaozerny LLC of the Varna region of the Chelyabinsk region was studied. The objects of research were the arable layer soil, water and feed produced on the territory of Zaozerny LLC of the Varna region of the Chelyabinsk region.

The data presented in table 1 showed that in the soils of the studied agroecosystem, the iron and copper contents exceed the MPC by 44.08% and 26.60%. The concentration of ecotoxicants — nickel, lead, and cadmium in the soils of Zaozerny LLC turned out to be 14.41 higher than the MPC; 16.95 and 29.87%, respectively, by elements.

A study of the water sources of Zaozerny LLC (natural body of water) for the content of heavy metals showed that the main toxic elements polluting the water are nickel, lead and cadmium.

In plant feeds, a high cadmium content was found, exceeding the MDL by almost 2.0 times. High concentrations of nickel in the soils of the economy also contribute to its intensive accumulation in plant feed, where the metal concentration exceeds MDL by 25.33%.

Studies have shown that environmental objects (soil, water and vegetable feed) of the household contain a high level of iron, copper and toxic elements - nickel, lead and cadmium, which leads to their abnormal content in the feed ration, and subsequently negatively affects the quality milk of cows contained in the conditions of technogenic agroecosystem.

During experiment I, the group of animals served as a control; it was used to judge the content of mineral elements in the blood and milk of cows contained in conditions of technogenic agroecosystems.

Cows of group II were given with food a 3% solution of water-soluble chitosan with a molecular weight of 38 kDa and a degree of deacetylation of 85% based on 2 ml / kg of animal body weight and the probiotic EM-Vita at a rate of 20 ml per animal, the preparations are used once per day for three 10-day courses with an interval of 7 days.

Group III cows were given acid-soluble chitosan with a molecular weight of 250 kDa and a degree of deacetylation of 88% based on 60 mg / kg of animal body weight 2 times a day for 5 days. Repeated use was carried out after 5 days.

Blood was taken for research before the experiment, and then on 20, 40 and 60 days from the date of use of the drugs. The content of mineral elements in the milk of cows was carried out at the same time.

The research results presented in table 2 showed that the level of iron in the blood of cows in the control group of animals during the entire experimental period does not change significantly and there are no significant differences with the initial values.

At the same time, with the use of water-soluble chitosan in combination with the probiotic EM-Vita, this indicator tends to decrease. So, on day 40 of studies, the level of iron in the blood was 24.12 ± 1.28 μmol / L, which is 24.98% lower than the initial level and 21.81% relative to the cows of the control group (in the prototype 12.62% and 12.25%, respectively). By day 60, the decrease in iron concentration was the highest and amounted to 45.10% and 44.79% (in the prototype - 37.86% and 34.63%).

It should be noted that on the 20th day of the experiment, the decrease in the level of iron in the blood of cows (group II) receiving water-soluble chitosan in combination with the probiotic EM-Vita was insignificant and amounted to 10.41%. This is probably due to the response of the body to the action of drugs aimed at protecting cells from the possible additional toxic effects of free iron ions by binding them to strong complexes with proteins.

A similar pattern is noted for copper. So, in the blood of cows of the control group, the copper concentration ranged from 19.73 ± 0.89 μmol / L to 20.46 ± 0.87 μmol / L. In the group of cows treated with water-soluble chitosan in combination with the probiotic EM-Vit, there is a significant decrease in copper concentration by 14.30% (40 days) and 36.06% (60 days) (in the prototype - by 9.18% and 24.14% on days 40 and 60, respectively).

The opposite tendency is characteristic for zinc, an element responsible for the immune system, the level of which in the blood of cows of the experimental group II significantly increased by 40 (35.12%) and 60 (39.88%) days of research in relation to the initial level (in prototype - 18.09% and 31.77%, respectively). In the cows of the control group, the increase in the level of zinc was 35.54% and 41.39%, respectively, for 30 and 60 days (in the prototype -16.00% and 30.38%). An increase in zinc concentration by the end of the observation period may be due to a decrease in blood levels of nickel and the formation of zinc thionine, which is a positive point during the correction of chitosan, water-soluble in combination with the probiotic "EM-Vita" with chronic intake of heavy metal salts in cows, how the synthesis of zinc thionins precedes and is mandatory for DNA synthesis during liver regeneration.

With the combined use of drugs (water-soluble chitosan and “EM-Vita”), the concentration of nickel in the blood of cows at the initial level of 11.33 ± 0.50 μmol / L on day 40 of the experiment decreases to 9.88 ± 0.41 μmol / L (in prototype - 10.34 ± 0.46 μmol / l) and 3.46 ± 0.12 - on day 60 (in the prototype - 7.53 ± 0.22 μmol / l). In cows of the control group, the nickel concentration in the blood ranged from 11.56 ± 0.43 μmol / L to 14.02 ± 0.69 μmol / L.

The very high adhesive properties of water-soluble chitosan in combination with the probiotic "EM-Vit" show in relation to lead and cadmium - an element with antimetabolic function. As the data in table 2 show, the lead content in cows of group II on the 20th day of the experiment increases relative to the background values by 18.14% (in the prototype - 4.66%). However, by 40 and 60 days of research, there was a significant decrease in lead concentration compared to the initial level by 31.86% (in the prototype - 26.69%) and 62.25% (in the prototype - 55.50%), respectively, by day . Apparently, the effective increase in the elimination of lead in the blood was influenced by a uniform increase in the level of zinc in the body of cows, which is a physiological antagonist of lead.

Blood loses most of the cadmium for a sufficiently long time, as evidenced by a sharp decrease in it in the blood only by 60 days to 0.28 ± 0.01 μmol / L at the initial level of 0.66 ± 0.03 μmol / L (in the prototype - to 0 , 47 ± 0.02 μmol / L with a background value of 0.59 ± 0.02 μmol / L). In the control group of cows, the concentration of cadmium in all periods of studies, on average, was 0.58 ± 0.02 μmol / L.

The analysis of the obtained results proved the significant effect of the combined use of water-soluble chitosan and the probiotic “EM-Vita” on the mineral composition of cow's milk (table 3).

By the end of the experiment (60 days), under the influence of chitosan water-soluble in combination with the probiotic EM-Vit, the iron content in cow's milk decreases by 17.27% compared to the background (in the prototype - 14.30%).

On 40 and 60 days of research, there is a decrease in the level of copper in the milk of cows of the II experimental group by 18.00% and 22.94% (in the prototype - by 15.05% and 18.74%, respectively, according to the days of research).

The zinc level in the milk of cows of the experimental group II significantly increased by the end of the experiment by 21.78% (in the prototype - by 10.08%).

In relation to toxic elements - nickel, lead and cadmium, the combined use of water-soluble chitosan and probiotic was manifested by high adhesive properties. A significant decrease in the level of these elements is observed at 40 and 60 days of research. The nickel concentration in the cows of group II was below the initial level in 2.08 and 5.33 times (in the prototype - 1.81 and 4.04 times); lead - 1.79 and 4.22 times (in the prototype - 1.15 and 3.21 times); cadmium - 2.17 and 5.50 times (in the prototype - 1.58 and 3.67 times), respectively, at 40- and 60 days of research. In the control group of cows, the content of nickel, lead and cadmium, on the contrary, continues to increase, compared with the background value.

Against the background of a decrease in milk of group II cows (water-soluble chitosan + "EM-Vita") iron, copper, nickel, lead and cadmium and an increase in the level of zinc, there is an improvement in the qualitative characteristics of milk - the content of total protein, casein, fat, lactose and skimmed milk powder residue (SOMO). In the milk of cows of the experimental group II, the total protein content increases by 17.24% (40 days) and 22.76% (60 days) (in the prototype - by 12.45% and 18.02%, respectively, on 40 and 60 days of the experiment ) In the milk of cows of the control group, this indicator does not significantly change.

The concentration of the main milk protein - casein, which is an indicator of the nutritional value of milk, rises by the end of the experiment in cows of group II by 31.44%, compared with the background (in the prototype - 21.55%). In this case, the level of casein in the milk of the cows of the control group increases slightly and does not have significant differences with the initial indicators.

The use of drugs had an effect on the fat content in the milk of experienced cows. The most significant increase (by 11.83% and 17.46%, respectively, on the 40th and 60th day of the experiment) of the fat content in milk of cows was observed with the use of water-soluble chitosan in combination with the probiotic “EM-Vita” (in the prototype, the increase in fat content in milk cows amounted to 9.06% and 11.04%, respectively).

Along with the increase in fat content in milk, there is a tendency to increase the concentration of dry skim milk residue. So, the level of SOMO in the milk of cows of the II experimental group at the initial level of 8.25 ± 0.14% by 40 and 60 days of research increases by 6.54% (in the prototype - by 4.86% and 5.22%, respectively) .

The results obtained and the patterns revealed indicate that the use of water-soluble chitosan in combination with the probiotic EM-Vita in conditions of technogenic agroecosystems significantly reduces the content of heavy metals (nickel, lead and cadmium) in blood and milk, and contributes to the balance of microelements such as zinc and copper; increases the concentration of total protein, casein, fat, lactose and COMO.

Thus, the advantage of the claimed method, in comparison with the prototype, allows us to recommend water-soluble chitosan with a molecular weight of 38 kDa and a degree of deacetylation of 85% from the calculation of 1.8-2.2 ml / kg of animal body weight and the probiotic "EM-Vita" from the calculation 18-22 ml per animal once a day for three 9-11-day courses with an interval of 6-8 days for use in animal husbandry to reduce the content of heavy metals in cow's milk.

With the introduction of water-soluble chitosan in combination with the EM-Vita probiotic in animal husbandry technology, economic efficiency is 5.22 rubles per ruble of costs.

Literature

1. Verotchenko, M.A. A system of measures to reduce the content of heavy metals in the trophic chain for agricultural enterprises in areas of technogenic pollution / M.A. Verotchenko, L.S. Gimadeeva, I.V. Gusev et al. // VIZh Recommendations. Dubrovitsy. - 2010 .-- 43 p.

2. Patent for the invention of the Russian Federation No. 23002109 “A method for reducing the level of nickel and lead in the blood and milk of cows of a technogenic province”, 2007 (Gertman A.M.).

3. Akhmetzyanova, F.K. The effectiveness of reducing the intake of heavy metals from feed into milk when sorbents are included in the diets of cows / R.G. Ilyazov, F.K. Akhmetzyanova, M.I. Gilemkhanov // Uchenye zapiski Kazan GAVM. - Volume 191. - Kazan, 2008 .-- S. 100-109.

4. Tairova, A.R. Environmental safety of livestock products with chitin / chitosan / A.R. Tairova, L.G. Mukhamedyarova / M-ly scientific.-practical. conf. “Scientific and technical creativity of youth - the path to a knowledge-based society”, M., - 2009. - S. 342-345.

Claims (1)

A method of reducing the content of heavy metals in cow's milk, including the use of chitosan, characterized in that the animals are given with feed a 2.7-3.3% solution of water-soluble chitosan with a molecular weight of 38 kDa and a degree of deacetylation of 85% based on 1.8- 2.2 ml / kg body weight of the animal and the probiotic "EM-Vita" at the rate of 18-22 ml per animal, the drugs are used once a day for three 9-11-day courses with an interval of 6-8 days.