Application of feedstuff treated with "hymizyme" polyenzymatic preparation in case of intoxication with heavy metals in animals and hens

FIELD: veterinary science.

SUBSTANCE: it is necessary to apply the feedstuff treated with "Hymizyme" polyenzymatic preparation by high-temperature fermentation technique in case of intoxication with cadmium and lead in animals and hens. The innovation enables to decrease the content of heavy metals in meat and poultry eggs.

EFFECT: higher efficiency.

8 ex, 3 tbl

 

The invention relates to the field of zootechnics, with the possibility of its application in veterinary medicine, in particular to the means of treatment and prevention of the excess amount in feed or poisoning animals with heavy metals, namely cadmium and lead.

From the large number of heavy metals of anthropogenic origin were the most dangerous cadmium and lead, which can enter the body of animals by air, water and food.

One of the features of the toxicity of heavy metals is that they have, as a rule, the cumulative effect, i.e. is able to accumulate in the body, and in most cases, these properties are endowed with cadmium and lead. The specificity of the toxic action of heavy metals is that, with its wide range of manifestations, these elements affect almost all organs and functional systems of the body, but the highest concentrations of cadmium and lead were observed in the liver and kidneys.

The allocation of heavy metals from the body occurs mainly in the feces and to a lesser extent from the urine, sweat, milk, saliva, bile and through the hair, but the greatest danger is the amount of heavy metals that has not withdrawn from the body and moves from animal products in the human body.

Known means for the treatment of the s poisoning of animals by heavy metals - this activated charcoal, laxatives and coating means (tannin, a decoction of flax seed, a solution of pectin), unithiol (Sdmicro, Ajarov, S. Antonov "Diagnosis of poisoning of animals". - M.: Agropromizdat, 1986; Dccallow "Pharmacology with recipes". - M.: Agropromizdat, 1986), sodium sulfide (Vgideo, V.a.novikov, Vahonicheva and other patent for invention №2178701, M., 2002) and others, However, ingestion of large doses of heavy metals these drugs for the treatment of animals are not sufficiently effective even after repeated application.

It is also known for the treatment of poisoning by cadmium and lead - sodium thiosulfate ("Veterinary encyclopedic dictionary". - M.: Soviet encyclopedia, 1981; "Medicinal product" / Directory. - M.: Russian book, 1994).

However, the repeated intravenous administration of sodium thiosulfate in large doses in case of poisoning of animals by heavy metals greatly increases the complexity of veterinary specialists and limits its practical use in the cattle industry, reduces the productivity of animals in the stress and side effects when you commit animals. In addition, sodium thiosulfate has a therapeutic effect in the first hours after the poisoning, the use of it at a later date is ineffective.

The task of the invention is to PR the change multienzyme preparation "Gelisim", intended for enzymatic processing of the concentrates, the use for the prevention and treatment of poisoning hens heavy metals. Preventive and therapeutic effects when fed to poultry and animal fodder grain, processed multienzyme preparation "Gelisim".

The use of a multienzyme preparation for the prevention and treatment of poisoning hens heavy metals has many advantages compared to therapeutic drugs that he used in animal feeding constantly, without negative effects on the body of the animal and its products (meat and eggs), but instead serves only to increase, and in addition do not spend extra money on medicines. In addition, the use of this drug will reduce the content of heavy metals in meat and eggs of birds grown in man-made regions of our country.

The mechanism for the protective effect of fermented feed in case of poisoning by heavy metals, obviously, is based on the improvement of metabolic processes in the body of animals and birds, which in turn leads to an increase in immunity, resulting in a better survival rate in chronic poisoning of animals, which is especially manifested in the early stages of the experiment, and later starts to happen correction is responsobitily reactions of an organism in response to excessive intake of cadmium and lead.

Multienzyme preparation "Gelisim" produced by the Foundation for the Development of economic reforms" (Kazan) and is intended for enzymatic processing of concentrated feed at elevated temperature in an easily digestible animal feed. "Gelisim" contains a set of hydrolytic enzymes, providing under the recommended conditions fast and deep cleavage feed nutrients in low-molecular digestible form, a complex of antioxidants, adsorbent and preservative. The product is designed for high temperature fermentation of concentrated feed. Ready food consists of 65-75% water and 25-35% of grain.

The procedure for the preparation of feed:

1. In mixer or other container for preparation of feeds pour cold water.

2. Dissolve the drug "Gelisim in 5-10 l of warm water (20-40° (C) the rate of 1 kg of the product per 1000 kg subject to processing of grain.

3. To make the dissolved drug in the mixer with water and stir.

4. Pour in mixer the grain coarsely.

5. Wet mesenko stir and heated to 70-100°C.

6. Leave to Mature for 1-2 hours.

Therapeutic and prophylactic efficacy multienzyme preparation "Himesama" had on white rats and chickens in acute and chronic complex priming animals with cadmium and lead. Observation zhivotnymi conducted during the entire experiment.

Example 1. The study of the prophylactic and therapeutic efficacy of the drug in rats. The experiments were carried out on white rats-males live weight 180-220 g, divided into two groups: the first group served as control and the second by experience. Animals orally was administered cadmium sulfate and lead acetate in sredneseriynoe dose LD50accordingly 350 mg/kg and 3000 mg/kg to Rats of the second group during the month prior to the production of experience and throughout the experiment were fed the grain, processed multienzyme preparation "Gelisim", and the first - past the entire production process but without making enzymes.

In the study of animals took into account the General condition, with respect to forage and water, the condition of the mucous membranes and hair coat, body weight, time of death or recovery of the animals. The results of the experiments for the study of therapeutic and prophylactic efficacy of the drug are shown in table 1.

Table 1

Therapeutic efficacy of multienzyme preparation "Gelisim" poisoning of white rats with heavy metals in the dose LD50.
Groups of animalsThe number of animals, goalTime of death, the day. In brackets the number of dead animals
onlysurvivedPalo
1. The seed cadmium and lead3015151(10), 2 (4), 3(1)
2. The seed cadmium and lead using fermented feed302372 (2), 3 (4), 4 (1)

The table shows that in case of poisoning white rats cadmium sulfate and acetate of lead in a dose of 350 mg/kg and 3000 mg/kg, respectively, the death of animals started at day 1 after the introduction of heavy metals, continued for 3 days. Just fell 15 white rats of 30, the survival rate was 50%.

When fed to laboratory animals of grain, processed multienzyme preparation "Gelisim", 30 poisoned white rats survived 23, i.e. to 76.7%. When compared with control animals clinical signs of poisoning were developed at a later date, namely, the 2nd day since the introduction of heavy metals, and were less pronounced.

Symptoms of poisoning was characterized by agitation, by refusing food and water, theresaknott coat, ataxia and weight loss. One rat (the first group) were killed 6 hours after drug injection. The condition of the surviving animals were visually normal during the first three or four su is OK. In the subsequent 14 days of changes in the clinical condition of the animals was not recorded.

The clinical examination of animals, which were fed by the grain, processed "Himesama", found that weight loss and other signs of poisoning were less pronounced in rats of the control group. Surviving animals looked quite actively, willingly took food and water.

Example 2. The study of therapeutic and prophylactic efficacy multienzyme preparation was carried out with feeding chickens of grain processed "Himesama"when priming with heavy metals. Birds were divided into two groups. Chickens in both groups asked for 22 days in bread boles sulfate cadmium and lead acetate at a dose of 52.5 mg/kg and 400 mg/kg body weight, respectively, which is 1/10 LD50respectively. The first group served as control. Birds of the second group before and during the whole time of the experiment received the grain, processed multienzyme preparation "Gelisim", and the first - past the entire production process but without making enzymes.

Determination of the clinical status of birds, as well as the study of blood on the content of erythrocytes, hemoglobin, leukocyte and leukocyte counts, total protein and its fractions, cholesterol and sulfhydryl groups, as well as phagocytic act shall want to make, conducted prior to the seed (background) and on the 10th day of control and experimental animals in comparison. All parameters were determined by standard methods.

During the experience were taken into account the content of heavy metals in the discharge of living, as well as the organs and tissues of dead and dead birds, which were determined by atomic absorption spectrophotometer "quantum-AFA" according to GOST 30178-96, using Methodical recommendations for spectral determination of heavy metals in biological materials and environmental objects.

The results of the experiments for the study of therapeutic and prophylactic efficacy of drug poisoning hens heavy metals at a dose of 1/10 LD50are shown in table 2.

Table 2

Therapeutic efficacy of multienzyme preparation "Gelisim" the daily seed hens heavy metals at a dose of 1/10 LD50.
Groups of animalsThe number of animals, goalTime of death, the day. In brackets the number of dead animals
onlyPaloCTR. slaughter
1. The seed cadmium and lead302821 (8), 3 (9), 12 (10), 11 (11), 1(12)
2. The seed cadmium and lead using fermented feed302822 (17), 3 (18), 5 (19), 8 (20), 9 (21), 1 (22)

The table shows that the daily seed chickens cadmium sulfate and acetate of lead in a dose of 52.5 mg/kg and was 400.0 mg/kg, respectively, bird deaths in the control group were started on day 8 after the introduction of heavy metals and continued for the next 5 days, and when the feeding of the birds of grain, processed multienzyme preparation "Gelisim", they began to die only on the 17th day. When compared with birds in the control group, in which clinical signs of poisoning in the form of a General depression, loss of appetite, therasense began to develop for 3-4 days, then in the experience of this occurred at a later date, namely 5-6 days since the seed.

With the simultaneous introduction into the body of chickens salts of lead and cadmium in a dose of 1/10 LD50on day 10 of the study, the hemoglobin content in the control decreased to of 73.5 g/l, which is 30% less than baseline data, erythrocytes to 4.53·1012/l or 26%, and leukocytes to 31.5·109/l or 10%, and experienced hens content of erythrocytes was the 5.45·1012/l, leukocytes 33,2·109/l and hemoglobin of 90.3 g/l, is below the background data, respectively, 11%, 5.2% and 14%. In the structure of leucoformula observed a statistically significant increase in basophils by 20% in comparison with the background and statistically significant increase of eosinophils. Throughout the experience, there is a decrease of total protein in serum, for example, on the 10 day experience in control chickens this indicator was significantly different from the background value and amounted to 43.1 g/l or decreased by 27%, while in the experimental birds these data were respectively 48,4 g/l and 18%. In the fractional composition of the protein in the control birds was observed a significant decrease in the number of albumin from 21.2 g/l at the beginning and to 13.4 g/l to 10 days of research, as well as a significant reduction of the level of globulin fractions. Experienced hens, these figures have changed slightly.

The control chickens, poisoned by heavy metals in a dose of 1/10 LD50, there was a slight decrease in phagocytic activity by 10% in comparison with the baseline data for 10 days from the beginning of the seed. Experienced chickens this index was little changed.

The number of sulfhydryl groups on the 10th day of the experiment decreased in chickens of the control group with 2,500 to 1825 mmol/100 ml of blood or 27%, while in the experimental group, respectively 2025 mmol/100 ml of blood or 19%. These changes have been reliable.

Thus, studies have shown that long-term is rancesca poisoning mixture of salts of cadmium and lead in doses 1/10 LD50occur statistically significant changes in leukotriene, hemoglobin, total protein and protein fractions, phagocytic activity and sulfhydryl groups of blood, which were more pronounced in birds of the control group compared with the experimental chickens.

In the study of heavy metals in the discharge, it was found that the use of the enzyme preparation contributed their best hatchability from the body of birds, for example, the average content of cadmium and lead in the litter of birds from the experimental group was higher than control, respectively, 1.5 and 1.2 times. It should be noted that in the last 5-7 days of life experienced hens heavy metals was to be distinguished from litter more than 2-3 times, than in the early days of the experience, for example, the average daily emissions of cadmium in the end the experience was 530-630 mg/kg litter, and in the early 90-120 mg/kg, and lead, respectively 930-1001 mg/kg and 100-130 mg/kg a Greater allocation of heavy metals from the droppings of chickens from the experimental group compared to the control indicates a positive effect of feed processed multienzyme preparation "Gelisim", on the body of birds. The reason for the more effective removal of heavy metals from the litter when fed fermented feed is probably in this case a deeper cleavage is nutritional the x substances food to low molecular weight compounds, accordingly, in the environment of a growing number of functional groups able to bind heavy metals compared to non-hydrolyzed biopolymers.

To study the content of heavy metals in organs and tissues on day 10 of the experiment was performed a control slaughter of birds, the results of which showed that the highest content of cadmium was observed in the liver of 27.5 mg/kg in the experience and 47.9 mg/kg in control, the stomach - 6,97 mg/kg and 24,71 mg/kg and kidneys are 40.2 mg/kg and 52,54 mg/kg, and lead in the stomach - 76,75 mg/kg and 145,96 mg/kg and liver - 32.1 mg/kg and 47,27 mg/kg, respectively. Muscle content of cadmium was experienced hens of 1.07 mg/kg and in control of 3.05 mg/kg or 2.85 times less, and lead respectively of 8.27 mg/kg and 12.0 mg/kg or 1.45 times less.

The dead animals were also studied heavy metals in organs and tissues, resulting in it was also found highest levels of cadmium in the liver - 56,9 mg/kg in the experience and 62.6 mg/kg in control, the stomach - 46,5 mg/kg and 53.2 mg/kg) and kidney - 43,56 mg/kg and 56,84 mg/kg, and lead in the stomach - 162 mg/kg and 186,5 mg/kg and liver -114,6 mg/kg and 128,4 mg/kg, respectively. Muscle content of cadmium was experienced hens of 1.57 mg/kg and control 3.1 mg/kg or 1.98 times less, and lead, respectively RS 9.69 mg/kg and 12,78 mg/kg or 1.32 times less.

Thus, the results indicate salts of heavy metals in organs and tissues of hens in the control slaughter is the dead showed processing feed multienzyme preparation contributed to the reduction of their deposits in organs and tissues of experimental birds compared to control.

Example 3. Given that in natural conditions, possibly infecting animals and birds is in smaller quantities, it was decided to reduce the dose of heavy metals, therefore preserving the principle of construction experience, similar to that described in example 2 chickens in both groups (control and experiment) were asked for 120 days in bread boles sulfate cadmium and lead acetate respectively in a dose 26,25 mg/kg and 200 mg/kg body weight, which is 1/20 LD50.

Determination of the clinical status of birds, as well as the study of blood on the content of erythrocytes, hemoglobin, leukocyte and leukocyte counts, total protein and its fractions, cholesterol and sulfhydryl groups, as well as phagocytic activity, was carried out before the beginning of the seed (background) and at 10, 20, 30, 60,90 and 120 days in the control and experimental animals in comparison.

During the experience were taken into account the content of heavy metals in fluids, organs and tissues of dead and dead birds.

The results of the experiments for the study of therapeutic and prophylactic efficacy of drug poisoning hens heavy metals in the dose of 1/20 LD50given in table 3.

T the blitz 3

Therapeutic efficacy of multienzyme preparation "Gelisim" the daily seed hens heavy metals in the dose of 1/20 LD50.
Groups of animalsThe number of animals, goalTime of death, the day. In Brackets the number of dead animals
onlyCTR. slaughterPalosurvived
1. The seed cadmium and lead30426-1 (18), 2 (19), 4 (20), 10 (21), 6 (22), 1 (25), 1 (28), 1 (35)
2. The seed cadmium and lead using fermented feed30123151 (25), 1 (26), 1 (27)

The table shows that the daily seed chickens cadmium sulfate and acetate of lead dose 26,25 mg/kg and 200.0 mg/kg, respectively (1/20 LD50death of the birds of the control group was started on the 18th day after the introduction of heavy metals and continued for the next 18 days, and when the feeding of the birds of grain, processed multienzyme preparation "Gelisim", they began to die only on the 25th day and killed 3 of the head compared with twenty-six in the control. When compared with birds in the control group, whose clinical p is snake poisoning in the form of General oppression reduce appetite, therasense began to grow for 5-7 days, then in the experience of this occurred at a later date, namely, on the 10-12 day since the introduction of heavy metals. In addition, it should be noted that females experienced group of clinical signs of poisoning were observed within 2-3 weeks, during which 3 birds were killed and the surviving chickens overall health and blood started to come in norm to 2-3 months from the beginning of the experiment, despite the continuation of oral administration of heavy metals in the dose of 1/20 LD50.

With the introduction of drugs in doses 1/20 LD50the control chickens was observed a decrease in the hemoglobin content in the blood. With 10 days this decrease was significantly and amounted to 83 g/l, which is 21% less than the background values, the content of the erythrocytes also significantly decreased to 4,96·1012/l or 19%, and leukocytes, respectively, to worth 32.55·109/l or 7%. Experienced hens content of erythrocytes was 5,75·1012cells 33,6·109and hemoglobin of 99.7 g/l, which is below the background data, respectively, 6%, 4% and 5.1%. 10 day experience in the structure of leucoformula both groups observed a statistically significant increase in basophils by 10% and eosinophils - 5% compared with the background data.

Throughout the experience, there is a decrease in total proteins is in the serum. 10 day experience this reduction in control chickens was significantly different from the background value by 16% to 49.6 g/l, while experienced this decline amounted to only 11% 52,5 g/l In fractional protein a decrease in the number of albumin, which is reliable and different from background values in the control birds at 24%, and $ 16.1 g/l, as well as significant decrease of the content α-, β- and γ-globulin. Reducing protein fractions in the experimental chickens wore unreliable character.

The control chickens, poisoned by heavy metals in the dose of 1/20 LD50observed false decrease in phagocytic activity by 7% compared with the background data for 10 days from the beginning of the seed. Experienced chickens this indicator remained at the same level.

The number of sulfhydryl groups on the 10 day experience decreased in chickens of the control group with 2500 2025 mmol/100 ml of blood or 19%, while in the experimental group, respectively, to 2265 mmol/100 ml of blood or 9.4%. These changes have been reliable.

On the 20th day all the studied parameters tended to decrease, both in experience and in control, but in laying hens of the experimental group it was less pronounced compared to control.

The General condition and the analyzed indices of blood in the surviving chickens from the experimental group started is RichEdit normal to 60-90 day, despite the continuation of oral administration of heavy metals in the dose of 1/20 LD50.

Thus, studies have shown that long-term chronic seed chickens salts of cadmium and lead in doses equal to 1/20 LD50, was statistically significant changes in hemoglobin, total protein and protein fractions, phagocytic activity and sulfhydryl groups of blood, which were more pronounced in birds of the control group in comparison with the test.

In the study of heavy metals in the discharge, it was found that the use of the enzyme preparation contributed their best to the excretion of birds. So the average content of cadmium and lead in the litter of birds from the experimental group were lower than in the control, respectively, in 3,42 and 3.02 times. It should be noted that in the last 15 days of the experiment in the experimental chickens average daily emissions of cadmium from litter was 23,81 mg/goal, and at the beginning of the experience 9,01 mg/goal or 2.6 times more than in the first days of the study, and lead, respectively - 64,9 mg/goal of 20.85 mg/goal or 3.11 times. Greater allocation of heavy metals from the droppings of chickens from the experimental group, compared to control, especially in the last days of the experiment, demonstrates the positive medical effects of feed processed preferment the drug "Gelisim", on the body of birds.

To study the content of heavy metals in organs and tissues on day 10 of the experiment was performed a control slaughter of birds, the results of which showed that the processing of the feed multienzyme preparation contributed to the decrease in the deposition of heavy metals in organs and tissues of experimental animals. In particular, the highest levels of cadmium were observed in the liver - 19,27 mg/kg in experience and 31,98 mg/kg in the control kidney - 33,64 mg/kg and 34,52 mg/kg and the stomach is 3.3 mg/kg and was 7.45 mg/kg, and lead in the stomach - 41,14 mg/kg and 93,14 mg/kg, kidney - 33,64 mg/kg and 42,66 mg/kg and liver - 27,61 mg/kg and 39,84 mg/kg, respectively. Muscle content of cadmium was experienced chickens 0,415 mg/kg and control 1.25 mg/kg or 3 times less, and lead - 5,14 and 8,87 mg/kg or 1.72 times less. On the 20th day of the experience was again performed a control slaughter, which showed that the content of heavy metals in organs and tissues grown in experience and in control, but in laying hens of the experimental group of cadmium and lead in organs and tissues was lower than in control: in the liver of cadmium 1.5 and lead 1.4 times, in the kidneys at 5% and 25%, in the stomach 1.9 and 2.1 times, respectively.

The dead animals were also studied heavy metals in organs and tissues, resulting in the greatest amount of cadmium was detected in the liver - to 57.1 mg/kg in experience and 63,46 mg/kg in control, in the muscle tissue of the stomach - to 43.5 mg/kg is 55,18 mg/kg) and kidney - 39,45 mg/kg and 60,18 mg/kg, and lead in the stomach - 159,21 mg/kg and 192,16 mg/kg and liver - 105,35 mg/kg and 119,21 mg/kg, respectively. In muscle tissue cadmium content averaged in the experimental chickens 1,51 mg/kg and control 2,95 mg/kg or in 1,95 times less, and lead, respectively, to 9.91 mg/kg and 14,62 mg/kg or experience 1.47 times less than in control.

Thus, we can conclude that the processing of the feed multienzyme preparation "Gelisim" contributed to the decrease in the concentration of heavy metals in organs and tissues of experimental birds compared to control.

In the study of heavy metals in the eggs was made the following conclusion: the control chickens cadmium averaged 0,462 mg/kg, and experienced is 0.260 mg/kg, and lead, respectively 6,66 mg/kg and 3,967 mg/kg, therefore, the use of enzyme preparation in laying hens of the experimental group was allowed to lower in eggs cadmium content in 1,78 times, and lead 1.68 times compared to control.

The mechanism for the protective effect of fermented feed in case of poisoning by heavy metals, obviously, is based on the following factors:

first, as noted above, the contents of the intestines of the bird has a higher complexes (binding) properties in relation to heavy metal ions by increasing the amount of low molecular weight compounds with functional the mi groups compared to only partially hydrolyzed biopolymer;

secondly, the increase in biogenic features feed after fermentation leads to increased immunity and protective properties of the organism, thanks to the animals and birds are easier to tolerate the initial period of acute poisoning, after which the body has time to start the generation of additional factors protect against toxicant (the increase in the content of sulfhydryl groups in the experiment compared to control);

thirdly, detoxification feed during high-temperature fermentation (digestion anti-nutritional substances, mycotoxins etc) leads to the focus of the protective properties of an organism to struggle with only one toxicant - heavy metals, and therefore is a more effective protection of the body;

fourthly, the birds of the experimental group under the influence of feed processed multienzyme preparation "Gelisim"probably starts to happen correction adaptive reactions of the organism in response to excessive intake of cadmium and lead.

Thus, studies have shown that the use of feed processed multienzyme preparation "Gelisim", in contrast to existing medicines, will save the cost of medicines, will facilitate the work of the veterinary specialists, will not create excessive stress and along with it will not be necessary shall Timoti diagnosis of poisoning of animals and birds, what is lost staff time and laboratory reagents, in addition, the use of this drug will reduce the content of heavy metals in meat and eggs of birds bred in the industrial regions of the country with their high concentration.

The use of feed processed multienzyme preparation "Gelisim" technology of high-temperature fermentation, as a therapeutic and preventive agent for excessive content in feed or poisoning animals and chickens heavy metals - cadmium and lead.



 

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1 ex, 1 tbl

FIELD: poultry science.

SUBSTANCE: the present innovation deals with visual evaluation in chicken followed by testing them by the value of bioelectric potential. Chickens with bioelectric potential being significantly higher against average values are considered to be stress-resistant ones and those with bioelectric potential being significantly lower against average values in concrete population are concluded to be stress-sensitive ones. The method is very simple in its implementation and efficient for large-scale selection in poultry on stress-resistance.

EFFECT: higher efficiency.

1 cl, 2 dwg, 2 ex, 4 tbl

FIELD: animal science.

SUBSTANCE: the suggested biopreparation named "Lactobifadol" should be applied 1 mo before the impact of the mentioned stress factor at the dosage of 100-200 mg/kg body weight, daily. Application of "Lactobifadol" enables to normalize physiological body state and decrease the losses of meat production.

EFFECT: higher efficiency of prophylaxis.

1 ex, 2 tbl

FIELD: animals science.

SUBSTANCE: the present innovation deals with intramuscular injection of sodium salt preparation cloprostenol 30-45 min before placing at the dosage of 750 mcg/animal. The method provides increased reproductive function, enhances sexual reflex, increases the volume of ejaculate, concentration, activity and quality of spermatozoa.

EFFECT: higher efficiency of breeding.

2 ex, 3 tbl

FIELD: animal science.

SUBSTANCE: one should measure bioelectric potential in biologically active centers N 5, 7, 11, 41, 44 to establish its average value and at its value being above 48 mcA one should conclude upon high growth energy in calves. The application of the suggested method enables to predict potential growth energy in calves beforehand.

EFFECT: higher efficiency and accuracy of evaluation.

2 ex, 1 tbl

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