Scientifically based method for correcting the metabolic processes of highly productive cows in a biogeochemical province

. The present study describes the features of the metabolic processes correction in cows with clinical signs of microelementosis (hypocobaltosis, hypocuprosis, rumen acidosis) in the biogeochemical province of the Chelyabinsk region. Study of the anamnesis showed that the deficiency of essential microelements (copper, cobalt, zinc, manganese), excess of iron and the presence of heavy metals (nickel) in environmental objects (soil, water, feed), in the blood of animals, deficiency of sugar, vitamins and low sugar-protein ratio in the feed ration caused the metabolic disorders and diseases of non-contagious etiology (hypocobaltosis, hypocuprosis, rumen acidosis) in the body of highly productive cows. Presence of metabolic disorders was confirmed by the results of morpho-biochemical studies of the blood of cows. In accordance with the purpose of the work, two groups of cows (experimental and control) of the first lactation trimester, 10 heads each, were formed using the principle of analogues. Metabolic disorders in the experimental group of cows were corrected using a complex daily therapy that included beet molasses (1.5 kg in a 1:3 dilution), sunflower cake (1.5 kg), I-sak 1026 (YEA-SACC1026) yeast culture at the rate of 1 g per 1 ton of planned milk (5-7 g per head per day) and “targeted” mineral premix (salts of cobalt sulfate (760 mg), copper sulfate (570 mg), manganese sulfate (650 mg), zinc sulfate (450 mg)). The mineral enterosorbent Vitartil (0.1 g/kg of body weight) was applied daily, in cycles of 15 days with an interval of 15 days. The duration of the experiment was 45 days. Corrective treatment was not performed in the control group of animals. The level of essential microelements in the blood

complete normalization of the parameters characterizing all types of cow metabolism, so it is necessary to continue the treatment.

Introduction
In order to increase the efficiency and competitiveness of livestock industries a number of serious problems must be solved, the main of which are insufficient supply of feed, its low quality and energy intensity, imbalance of diets for basic nutrients, biologically active substances and mineral elements.As for the latter, macro-and microelements play an extremely important role in regulating the intake of feed and water by animals, ensuring digestion, growth and development, formation and maintenance of health, high productivity and ability to reproduce [1].Excessive or insufficient intake of mineral substances disrupts the assimilation and dissimilation life processes, leads to deep, chronic metabolic disorders of the body's systems.In farms located in the biogeochemical provinces of the Southern Urals, with an excess or deficiency of certain macro-and microelements, non-contagious animal pathology is widespread.It is caused by metabolic disorders and includes specific diseases (osteodystrophy, microelementoses, hepatitis, hepatosis, urolithiasis, nephorosis, mycardosis, endometritis and others), which lead to significant economic damage [2].These pathologies are caused not only by the deficiency of essential microelements, such as cobalt, manganese, zinc, and others; a significant role in etiology belongs to antagonist elements, which are found in excess in the environmental objects of biogeochemical provinces and in the feed ration and animal body, accordingly [3].Dispensary examination of animals on certain territories shows that significant deviations of metabolic processes are detected in highly productive cows due to a deficiency of energy substances that are necessary for the formation of milk, a low level of micro-and macroelements and vitamins in the diet.A serious disruption of metabolic processes is caused by salts of heavy metals and iron, which are found in excess in the feed ration.A number of researchers are studying the effect of mineral inorganic and organic feed additives on the body of cattle [4][5][6][7].According to AR. Rabiee [8], JL.Siciliano-Jones [9], K. Dhakal [10], organic micronutrient supplementation can improve productivity and reproduction in lactating dairy cows.Dairy cattle's breeding in the indicated territories has its own characteristics, since traditional schemes for the normalization of metabolic processes have a low therapeutic effect.The purpose of this study was to correct the metabolic processes in highly productive cows in order to increase milk productivity and to treat a number of diseases of non-contagious etiology (hypocuprosis, hypocobaltosis, rumen acidosis) in a biogeochemical province.

Materials and methods
"Sarafanovo" agricultural production cooperative of the Chebarkulsky district of the Chelyabinsk region was chosen for the research.The land is located in the zone of emissions from the metallurgical complexes of the cities of Zlatoust, Miass, Chebarkul and is a biogeochemical province in which environmental objects (soil, water sources, fodder crops) contain a high level of iron and nickel with a pronounced deficiency of cobalt, manganese, copper, zinc.
The research was performed in three stages.At the first stage, environmental objects (soil, water sources, feed) of the studied farm were subjected to local monitoring for the content of toxic and essential elements.The microelement composition in the studied objects was determined by the atomic absorption method using an AAS-3 spectrophotometer.
At the second stage of scientific research, the influence of some essential microelements deficiencies (zinc, cobalt, manganese, copper) and an excess of salts of heavy metals (iron E3S Web of Conferences 462, 01008 (2023) AFE-2023 https://doi.org/10.1051/e3sconf/202346201008and nickel) on metabolic processes in the animal body was studied.These microelements play an important role in numerous enzymatic processes, in regulation of the metabolism of carbohydrates, proteins, lipids, minerals, in maintaining the activity of the antioxidant system, and prevent the development of anemia.Dispensary clinical study of animals was carried out according to the generally accepted method adopted in veterinary practice.794 heads of cows in the first 100 days of lactation were subjected to clinical study.Morphobiochemical parameters of blood were studied by unified methods given by I.P. Kondrakhin et al. and accepted in veterinary practice.The obtained data were compared with the normatives.
Taking into account the chemical content of the diet and the level of essential microelements in the blood of animals, a correction was carried out at the third stage of the experiment.In accordance with the purpose of the work, two groups of cows (experimental and control) of the first lactation trimester, 10 heads each, were formed using the principle of analogues.The control group of animals did not undergo corrective treatment; so the diet accepted in the agricultural enterprise was used.Based on the results obtained, beet molasses (1.5 kg diluted 1 to 3), sunflower cake (1.5 kg), I-sak 1026 (YEA-SACC1026) yeast culture at the rate of 1 g per 1 ton of planned milk (5-7 g per head per day), "targeted" mineral premix, which included salts of cobalt sulfate (760 mg), copper sulfate (570 mg), manganese sulfate (650 mg), zinc sulfate (450 mg) were daily introduced into the diet of the experimental group of animals.Given the high content of nickel and iron in environmental objects, the mineral sorbent Vitartil was used once a day in the amount of 0.1 g/kg of animal body weight in cycles of 15 days with an interval of 15 days.Enterosorbent and microelements were given separately.The duration of the experiment was 45 days.Statistical processing of the obtained data was carried out by calculating the average statistical error and Student's t-criterion.

Results
In the course of laboratory studies, it was found that the content of toxic elements (nickel and iron) significantly exceeded the MPC (maximum permissible concentration) and MAL (maximum allowable level) in all soil samples from various fields, as well as water sources and fodder crops of the "Sarafanovo" agricultural production cooperative of the Chebarkulsky district of the Chelyabinsk region.The content of copper, zinc, cobalt and manganese in external environmental objects was characterized by their pronounced deficiency.The results of the study of environmental objects for the content of chemical elements are presented in Table 1.According to the data from Table 1, serious deviations of the maximum allowable concentration were revealed.Thus, in soil samples taken from various fields of the farm, on which fodder crops were grown (barley, corn for silage, a mixture of annual grasses for haylage), a significant excess of iron content was revealed, exceeding the MPC by 4.3; 3.6; 3.2 times respectively.The amount of vital elements (copper, zinc, cobalt) was less than the MPC values.The amount of copper was by 71.4,69.8 and 27.6% below the MPC in soil samples obtained in the areas of cultivation of fodder barley, silage crops and haylage annual grasses, respectively.The content of zinc, cobalt, and manganese was reduced in all soil samples of the studied areas.At the same time, we noted a high nickel content, which exceeded the MPC by 21.4, 91.4 and 7.2% in soil samples of grain crops, silage and haylage, respectively.Toxic-chemical analysis of the farm water sources revealed the following changes.In water samples taken from the water tower, the level of iron was 60% lower than the MPC values, while in the drinking bowls of livestock buildings it was 4.7 times higher.The amount of copper, zinc and cobalt was significantly less than the MPC values: copper by 98.3; zinc by 70.0 and cobalt by 94.5%, respectively.
It was found that iron content exceeded the norm by 50.0, 46.7 and 39.7% in samples of fodder crops (barley), corn silage and haylage, respectively.The amount of microelements (copper, zinc, cobalt) was significantly lower than the MAL value.Exceeding the MAL for nickel was found in samples of all fodder crops, for example, the amount of nickel exceeded the MAL by 62.3% in concentrates, by 31.3% in corn silage by and by 29.7% in haylage, respectively.Thus, the obtained data convincingly indicate that the land use of this farm is a biogeochemical province.An analysis of the feed ration of cows revealed an acute deficiency of sugar, microelements, vitamins, a low sugar-protein ratio and high level of nickel and iron.This was the reason for the development of metabolic disorders.Target organs, such as the liver, which is the central organ of metabolism, and the kidneys, can be damaged by the toxic effects of heavy metal salts.At the same time, changes of functional state of these organs lead to a violation of metabolic processes.
A clinical examination of sick animals showed a decrease in skin elasticity, dull hair, decreased rumen motility, sagging of the spinal column in some animals, cracking in the joints, cachexia, liver tenderness and anemic mucous membranes.During the clinical examination of 794 heads, it was found that 36.6% of animals had disorders of the musculoskeletal system, 12% -microelementosis, 18% -liver disease (hepatitis, hepatosis), 2% -kidney disease (nephritis, nephrosis), 2.5% -lesions of the cardiovascular system (myocarditis, myocardosis), 5.5% -cicatricial digestion disorders (rumen acidosis).Thirty animals had several diseases.
The level of erythrocytes and leukocytes was reduced by 21.4 and 33.4%, respectively, with a simultaneous decrease in hemoglobin content, the level of which was 16.6% below normal.The simultaneous decrease in hemoglobin and the total number of erythrocytes suggests that anemia was developing in the body of cows.Thus, the hematopoietic process was disturbed and hypochromic anemia developed in sick animals.During the experiment, the animals of the experimental groups showed activation of hematopoietic processes, the most significant activation occurred on the 45th day of the experiment.The application of correction increased the number of erythrocytes by 29.9% (P<0.01),leukocytes by 39.5% (P<0.01),hemoglobin by 15.5% (P<0.01) in the blood in of cows from the experimental group in comparison with animals of the control group.The content of chemical elements in the blood of animals is presented in Table 2.According to the data presented in Table 3, the background content of zinc, cobalt, manganese and copper in the blood of cows of the experimental and control groups was reliably lower than the standard values by 30.5; 85.1; 60.0; 45.0% respectively.Iron and nickel act as antagonists, preventing the normal absorption of cobalt salts.A high level of detoxification of iron and nickel salts was noted in all periods of the experiment, but the most significant results were obtained on the 45th day of the experiment.Thus, the complex therapy led to a significant decrease in the level of iron (-9.9%) with a reliability of P<0.05 in the experimental group.This effect, in our opinion, may be associated with the sorption properties of the zeolite, since the level of nickel salts was reliably lower by 18.1 (P<0.01) and 23.8% in comparison with the control values and background indicators, respectively.At the same time, levels of zinc, cobalt, copper and manganese increased.Thus, the content of zinc increased by 34.1% (P<0.001),cobalt by 53.8% (P<0.001),copper by 28.8% (P<0.001),manganese by 55.5% (P<0.01) in comparison with the control group of cows.The results of the study allow us to conclude that Vitartil had sufficiently high ion-exchange properties, the micro-macroelements included in its composition provided compensatory regulation of mineral compounds metabolism.The results of the protein metabolism are presented in Table 3.According to the data obtained, the level of total protein and albumin in the blood serum of cows was reduced by 12.0 and 0.9% relative to the normative data.At the same time, the level of proteins of the globulin class, such as β-globulins and γ-globulins, increased by 67.8% and 20%, respectively, compared with the normative data.These results indicate a possible impairment of protein synthesis in the liver.This is confirmed by the high activity of the transamination enzymes AST and ALT.According to the data, the level of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in cows from the experimental group was higher than the norm by 25.0 and 12.9%.Elevated level of these enzymes indicates liver cells damaging.The level of urea also increased by 20.9% in experimental cows, which may be associated with kidney damage by toxic elements.The ongoing metabolic correction showed a positive trend throughout the entire experimental period, with the most significant results on the 45th day of the experiment.

Days of study
The level of total protein and albumin fraction of serum proteins in the blood of cows in the experimental group was by 9.7 (P<0.05) and 19.7% (P<0.05)higher compared to the control group.On the 45th day of therapy, the content of α-globulin increased by 13.5% (P<0.01), while the amount of β-globulin and γ-globulin decreased by 34.0 % (P<0.001) and 9.1% (P<0.05).
Thus, the therapy led to the normalization of liver function, which was manifested in a decreased activity of transamination enzymes and reduced level of urea.During hepatocytes damage cytosolic fractions of ALT and then mitochondrial AST are released and enter the bloodstream.By the end of the experiment, the level of AST and ALT decreased by 19.6 % (P<0.01) and 9.9% (P<0.05)relative to the animals of the control group.The end product of nitrogen metabolism is urea, the level of which by the end of the correction in the experimental group of cows was 10.9% lower than the control values (P<0.05).This indicates the normalization of the functional state of the excretory system of cows in the experimental group.
Cicatricial digestion in cows plays an important role in the metabolism of complex carbohydrates, as well as in the synthesis of vitamins and amino acids.Violation of these processes can lead to various non-contagious diseases and a decrease in milk production [11,12,13].The study of parameters associated with cicatricial digestion and carbohydrate metabolism highly productive cows allows a deeper understanding of the causes of some metabolic diseases and helps to solve these problems in veterinary practice.
Analysis of the background parameters of the rumen indicated a decrease in the pH of the rumen by 13.7%, the number of ciliates by 73.0% and total concentration of VFAs by 28.8%, respectively.The content of acetic and propionic acids was by 4.4 and 10.7% below the standard values, and the content of butyric and isobutyric acids was higher than the recommended values by 79.1 and 90.1%, respectively.Hypoglycemia in cows, accompanied by a decrease in glucose levels by 41.3% below normal and increased activity of 1,6diphosphate aldolase, indicates disorders in carbohydrate metabolism.This may points out to a violation of biochemical processes in the rumen and changes in carbohydrate metabolism.Excess of lactic and pyruvic acids by 40.4 and 30.1% may indicate the predominance of the anaerobic pathway of glycolysis.As a result of the correction, the parameters of cicatricial digestion and metabolism of carbohydrate compounds were normalized, which is reflected in Table 4.As a result of the correction, the reaction of the rumen environment in the experimental animals was 9.6% (P<0.05)higher than that in the animals of the control group.An increase in the number of protozoa in the rumen by 2.9 times (P<0.001)compared with the control resulted in an increase in the amount of volatile fatty acids by 29.9% (P<0.01).The proposed correction method led to an increase in acetic acid by 7.2% (P<0.05),propionic acid by a decrease in isobutyric acid by 28.9 (P<0.01),butyric acid by 32.4 % (P<0.01) in comparison with control animals.In the experimental group of cows, the amount of isovaleriс, valeric and caproic acids increased by 54.5 (P<0.001),18.0 (P<0.01) and 50.0%(P<0.001),respectively.
These changes in the composition of acids indicate the normalization of digestive processes in the rumen and more efficient metabolism in experimental animals compared to the control group.The positive changes of the carbohydrate metabolism parameters after the correction are shown in Table 5.According to the results from the table, there were a reliable increase in blood glucose levels by 46.9% (P<0.01) and a decrease in the activity of 1.6 diphosphate aldolase by 35.9% in the experimental group of cows compared with the control group.This is a favorable factor that contributes to the glycolysis regulation in general.In addition, we observed a decrease in the level of lactic acid by 17.5% (P<0.01) and pyruvic acid by 12.9% (P<0.05) in cows in the experimental group.These data confirms that correction effectively normalized the metabolism and reduced the negative impact of lactic and pyruvic acids on the digestive processes in animals.
Summarizing the data obtained, it should be noted that in the experimental group of cows there was an increase in milk productivity by 18.1% compared to the control group, the reproductive ability was activated, since four heads out of 10 cows of the experimental group were ready for mating by the end of the experiment.

Discussion
Summarizing the data obtained, it is necessary to conclude that the land use of the agricultural production cooperative was located in the zone of a biogeochemical province of technogenic origin with a high content of nickel and iron salts in environmental objects (soil, water sources, fodder crops) and, consequently, in the feed ration, with a pronounced deficiency of essential microelements such as cobalt, copper, zinc, manganese.This was accompanied by violation of protein, carbohydrate, mineral metabolism, development of diseases of non-E3S Web of Conferences 462, 01008 (2023) AFE-2023 https://doi.org/10.1051/e3sconf/202346201008contagious etiology (hypocobaltosis, hypocuprosis, rumen pcidosis).The use of complex treatment of these pathologies is physiologically justified.Sunflower cake has a balanced composition, the protein content can be up to 40%, the energy value is higher than in standard feeds.The detoxifying properties of the mineral sorbent Vitartil, which has high sorption properties for nickel and iron, in combination with other components of corrective treatment, contributed to a decrease in the level of toxic substances, normalization of the liver and kidneys functional state, activation of the protein metabolism and an increase in the content of total protein and albumin together with the use of sunflower cake.
The ion-exchange properties of Vitartil made it possible: to normalize the exchange of copper, zinc, cobalt and manganese, thus activating the hematopoiesis, which normalizes metabolic processes, stimulates carbohydrate, protein, fat metabolism, increases resistance to adverse environmental effects; to stimulate the production of erythropoietin; to prevent the development of anemia [14,15].The composition of this zeolite includes more than 40 macro-and microelements, which led to an increase in the amount of essential microelements in the blood of the experimental group of animals.It is known that volatile fatty acids (VFAs) are formed in the rumen of ruminants under the influence of enzymatic processes.VFAs serve as one of the main energy materials for animals.The precursor of glucose in ruminants is propionic acid, which is absorbed in the rumen wall, enters the blood and goes further to the liver, participating in carbohydrate metabolism.Thus, there is a close relationship between the processes of cicatricial digestion and the metabolism of carbohydrate compounds.Milk production increases during lactation, this requires increased energy support.Carbohydrates are the main source of energy for breast cells and for the synthesis of lactose, which is the main carbohydrate component of milk.Carbohydrate deficiency during this period can lead to reduced milk production and inadequate nutritional intake for milkconsuming calves.High milk productivity of cows in the biogeochemical province is accompanied by profound changes in metabolic processes.The results of this experiment confirm the positive impact of the combined use of the components of corrective treatment on the biochemical parameters of blood and cicatricial digestion, especially on the 45th day of the experiment.Due to its chemical composition, Vitartil has antacid properties and is involved in neutralization of acidic products excess, which is formed in the rumen.In addition, it acts as a sorbent and helps to remove metabolic products, toxins, salts of heavy metals, and gases from the gastrointestinal tract.The activation of cicatricial digestion was facilitated by the introduction of beet molasses into the diet of the experimental group of cows.Beet molasses is a source of fast energy, it stimulates microbial activity, and is used to feed the microflora.The pharmacological properties of the yeast culture are determined by the presence of B vitamins, amino acids and minerals in its composition, which ensures the growth and development of animals, improve food absorption, activates intestinal transport functions, normalizes metabolic processes, has an immunostimulating effect, and contributes to the elimination of toxic elements.The ongoing therapy had a significant effect on the redistribution of VFAs in the contents of the rumen.It increased the level of acetic and propionic acids and a decreased the amount of butyric and isobutyric acids.Thus, beet molasses, I-sak 1026 (YEA-SACC1026) yeast culture and Vitartil activated the processes of digestion in the rumen of highly productive cows from the experimental group and had a positive effect on the parameters of carbohydrate metabolism.

Conclusions
In the conditions of a biogeochemical province, when environmental objects contain an abnormal amount of micro-, macroelements and salts of heavy metals, all types of metabolism in cows are disturbed.Diet errors, deficiency of carbohydrates and microelements, high levels of nickel and iron salts are the main etiological factors that trigger E3S Web of Conferences 462, 01008 (2023) AFE-2023 https://doi.org/10.1051/e3sconf/202346201008profound changes of digestion processes in the rumen, metabolism of carbohydrates, proteins, and minerals in highly productive cows.The proposed scheme for correcting the feed ration of animals by introducing beet molasses, I-sak 1026 (YEA-SACC1026) yeast culture, a "targeted" mineral premix and Vitartil not only lead to the normalization of blood biochemical parameters of carbohydrate, protein, mineral metabolism, but also improved the cicatricial digestion and clinical status of animals, and provided high rates of milk production and reproductivity.

Table 2 .
The content of chemical elements in the blood of experimental cows, mg/l (M±m; n=10) Note here and below: M -arithmetic mean for the group; ±m -error of the arithmetic mean; n -the number of animals in the group; reliability: * -Р<0.05;** -P<0.01;*** -P<0.001

Table 4 .
Parameters of cicatricial digestion processes in cows after the therapy (M±m; n=10)

Table 5 .
Parameters of the carbohydrate metabolism during therapy (M±m; n=10)