Method for normalising blood antithrombin iii activity in newborn calves with iron-deficiency anaemia

FIELD: medicine.

SUBSTANCE: invention relates to veterinary science. A method involves the use of intramuscular injections of ferroglucinum 75 mg at 15 mg of iron per 1 kg of calf's body weight, once in a combination with feeding of glycopine 6.0 mg/day in the morning for 6 days starting simultaneously with injections of ferroglucinum 75.

EFFECT: method is high-effective in correction of blood antithrombin III activity.

1 dwg, 3 ex

 

The invention relates to veterinary medicine and biology, namely Hematology.

Analogues of the proposed method of correction activity of anti-thrombin III (AT III) in the blood of newborn calves with iron deficiency anemia does not exist.

In the literature there is information on the application of anemia in calves feroglobin (Internal non-infectious diseases of farm animals. Under. edit Vmenyaemogo. M: Agropromizdat, 1991. - 576 S.).

In the literature, there are also recommendations on the use of animals for the purpose of immunostimulation of glicerina (N-acetylglucosaminyl-N-acetyl-muramyl-alanyl-D-isoglutamine) (Andronov T.M., Pinegin B.V., goats I.G., Ustinov GI Application immunomodulator of glicerina for the prevention and treatment of animal diseases. Methodical recommendations. M., 2009. - 12 S.) But never before picopen and especially its combination with ferroaluminum was not used in newborn calves with iron deficiency anemia with the purpose of correcting existing weakening of the activity of at III in the blood that can cause thrombogenic risk in animals, dramatically worsening the course of anemia, weakening the reactivity of a living organism, often leading him to death.

The aim of the invention is to correct the activity of at III in the blood of newborn calves with iron deficiency anemia.

The essence of the proposed method lies in the fact that h is about for the correction of AT III in the blood of newborn calves with iron deficiency anemia intramuscularly once the solution is applied feroglobin-75 (1 ml), containing 75 mg of iron per 1 ml of solution at a ratio of 15 mg of iron per 1 kg of body weight of the calf and feeding of glicerina at 6.0 mg/day in the morning (diluted in a small amount (100,0-of 150.0 ml) of water or milk or colostrum, designed to feed calves) for 6 days, starting simultaneously with the injection feroglobin-75.

The method enables stable to normalize the activity of at III in the blood of newborn calves with iron deficiency anemia on the next day after the treatment, which will significantly improve the herd, to reduce mortality, increase the gain and get these animals healthy offspring.

The inventive method is carried out as follows.

The determination of the activity of at III in the blood of newborn calves with iron-deficiency anemia is as follows (Barkagan SS, Momot A.P. bases of diagnostics of disorders of hemostasis. M., 1999. - 214 S.).

The principle of the method is based on detecting the reduction of the activity (in the absence of heparin) plasma at III, which is determined by the ability of the investigated plasma to inactivate thrombin, to the pretreated sorbent heparin, then subjected to heat defibrinate and mixed with a standard amount of thrombin. After incubation of the mixture in it determine the residual coagulation activity of thrombin and level lowering activity and thrombin in percent from the norm estimate the activity of at III in the plasma.

Use the following reagents. 1. Tris-HCl buffer 0.05 M, pH 7.4. 2. A solution of thrombin in buffer: 0.2 ml of a solution should minimize 0.3 ml of 0.4% solution of fibrinogen for 15-16 seconds. The working solution of thrombin is prepared in a plastic or silikonizirovannoj tube, stored at room temperature and used in the test on the day of the study. 3. Sorbent heparin Genosorb-1 (firm "Technology Standard"). 4. A freshly prepared solution of fibrinogen at a concentration of 3.5-4.0 g/l, which is used for testing residual activity of thrombin. May be replaced with a solution of fibrinogen normal platelet-poor plasma nitrate dissolved with buffer 2 times. 5. Fresh nitrate platelet-poor plasma obtained from 6-8 healthy calves, or freeze brand, with the known activity of at III, which is used to construct a calibration graph.

The research material is nitrate platelet-poor plasma.

For the preparation of sorbed and defibrinating plasma in a test tube and mixed with 10 mg of sorbent heparin with 1.0 ml of the investigated plasma. Constantly shaking the test tube, mix the contents at room temperature for 8 min, excluding foam and sediment at its bottom. After that, the mixture was centrifuged for 2 min at 1000-1500 rpm Plasma in nadeshiko transferred to clean the wading is and defibrinated in a water bath at +56C for 6 min, then again centrifuged at 3000 rpm for 10 min, remove the supernatant portion and explore within the first 2 hours

The course definition. In test tube add 0.4 ml of the working solution of thrombin and heat it in a water bath at +37C for 2 minutes Then add 0.1 ml of the investigated adsorbed and defibrinating plasma include stopwatch. After 2 minutes (exactly!) 0.2 ml of the mixture is transferred into a test tube with 0.3 ml of a solution of fibrinogen (or diluted normal plasma), pre-warmed to +37C. Immediately include stopwatch and record the clotting time.

On the calibration curve to find the activity of at III in percent.

The calibration curve carried out as follows. The plasma of healthy calves are defibrinating, as described above. Then from preparing cultivation to prepare the calibration curve in accordance with table 1.

Table 1
The scheme of preparation of dilutions of plasma to construct a calibration curve for the determination of the activity of AT III
The number of breedingPlasma + bufferThe activity of AT III, %
1PL is ZMA without buffer 100
20.2 ml +0.2 ml50
30.1 ml +0.3 ml25

With each of the dilutions to determine the clotting time according to the above method. All the measurements were carried out three times, calculate the average result. When the calibration curve on the axis of ordinates delay the clotting time in seconds on the x-axis is the activity of at III in % in accordance with the above dilutions. A sample calibration curve is shown in figure 1.

A calibration curve for one type and one series of thrombin and buffer build once. Periodically inspects only the activity of the working solution of thrombin.

The normal activity of at III plasma is 85-115%.

For the correction decreased at III in the blood of newborn calves with iron deficiency anemia once assigned feroglobin-75 containing 1 ml of a solution of 75 mg of iron at the rate of 15 mg of iron per 1 kg of body weight of the calf, and the feeding of glicerina at 6.0 mg/day in the morning for 6 days, starting simultaneously with the injection feroglobin-75.

The method allows to normalize the activity of at III blood of newborn calves with iron deficiency anemia on the next day after the end l of the treatment, bringing it to the level characteristic of healthy animals, which increases the reactivity of the organism of the animal, treating the underlying disease, significantly reducing the risk of thrombotic complications and promoting normal growth and development of animals.

Example 1. Calf No. 74, the third day of life, the body weight of 27.0 kg, with iron deficiency anemia were examined in terms of the calf. The calf was taken and investigated blood activity score it at III, which was reduced to 75%. Calves assigned to 5.4 ml feroglobin-75 intramuscularly once and feeding diluted to 100.0 ml milk glicerina at 6.0 mg/day in the morning for 6 days, starting simultaneously with the injection feroglobin-75. The application of this correction allowed on the following day after graduation to raise the activity of at III to the normative values - 110,0%, eliminating the risk of CE weakening thereafter.

In further contain this calf was recommended on rational diet for the prevention of relapse of iron deficiency and attenuation of at III in the blood.

Example 2. Calf No. 82, the fourth day of life, body weight 31.5 kg, with iron deficiency anemia were examined in terms of the calf. The calf was taken and investigated blood activity score it at III, which was reduced to 78%. Calves assigned to 6.3 ml Fe is rogulina-75 intramuscularly once and feeding diluted in 150,0 ml milk glicerina at 6.0 mg/day in the morning for 6 days, starting simultaneously with the injection feroglobin-75. This allowed the next day after the final correction to raise the activity of at III to the normative values of 99%, eliminating the risk of weakening thereafter.

In further contain this calf was recommended on rational diet for the prevention of relapse of iron deficiency and weaker activity of at III in the blood.

Example 3. Calf No. 94, the second day of life, body weight 30.5 kg, with iron deficiency anemia were examined in terms of the calf. The calf was taken and investigated blood activity score it at III, which was reduced to 73%. Calves assigned to 6.1 ml feroglobin-75 intramuscularly once and feeding diluted in 150,0 ml milk glicerina at 6.0 mg/day in the morning for 6 days, starting simultaneously with the injection feroglobin-75. This allowed the next day after the end of correction to raise the activity of at III to the normative values of 107%, eliminating the risk of weakening thereafter.

In further contain this calf was recommended on rational diet for the prevention of relapse of iron deficiency and reduction in activity of at III in the blood.

The claimed method is tested on 149 newborn calves with iron deficiency anemia and showed 100% efficiency correction zeleste zitnay anemia and level of activity of at III in the blood.

The method of normalization of the activity of anti-thrombin III in the blood of newborn calves with iron deficiency anemia, including the use of a solution feroglobin -75 at the rate of 15 mg of iron per 1 kg of body weight of the calf intramuscularly once daily in combination with oral application of glicerina at 6.0 mg/day in the morning for 6 days, starting simultaneously with the injection feroglobin-75.



 

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