RU2403024C1 - Method of normalising plasminogen activator inhibitor in anemia of newborn calves and pigs - Google Patents

Abstract

FIELD: medicine, veterinary science.

SUBSTANCE: invention refers to veterinary science. The method involves application of Ferroglucin by 150 mg (2 ml) intramuscularly, two times every 10 days, Phosprenyl intramuscularly, two times by different syringes in different points, at first in dosage 0.08 ml/kg combined with an iron preparation, and for the second time in dosage 0.08 ml/kg five days after the first injection of ferroglucin and cresacine 3 mg/kg included in the feeding schedule for 8 days starting with the first injection of ferroglucin simultaneously.

EFFECT: method allows avoiding vascular complications in newborn calves and piglets with anaemia, reducing mortality, improving a livestock.

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Description

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

Analogues of the proposed method for normalizing the level of plasminogen activator inhibitor in newborn calves and piglets with anemia do not exist.

In the literature there is information about the use of ferroglukin in calves and piglets for anemia (Internal non-communicable diseases of farm animals. Edited by V.M. Danilevsky. M .: Agropromizdat, 1991. - 576 p.).

As an effective stimulant of the vital activity of a growing organism in modern veterinary medicine, a phosprenyl preparation based on phosphates of polyprenols obtained from needles is used (Deeva A.V., Salakhova Z.A., Lobova T.P. and others. Improving the safety and productivity of piglets when used phosprenyl and gamavit. // Veterinary medicine. - 2006. - No. 4. - p.13-15) and crezacin (a derivative of aroxyalkylcarboxylic acids), which is an effective stimulant for living organisms, increasing the survival of animals in adverse and extreme conditions, stimulating or normalizing life processes (Baimatov VN, Latypov MM The effect of krezacin on the state of the bird. Veterinary medicine, 2006, No. 11, p. 49-51).

However, never before has a treatment complex consisting of ferroglukin, fosprenil and crezacin been used in newborn calves and piglets with anemia in order to normalize the level of plasminogen activator inhibitor.

The aim of the invention is to increase the efficiency of correction of the level of plasminogen activator inhibitor in newborn calves and piglets with anemia.

The essence of the proposed method lies in the fact that to optimize the level of plasminogen activator inhibitor, newborn calves and piglets with anemia are prescribed ferrglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, phosprenyl intramuscularly, twice in different syringes at different points, the first time 0.08 ml / kg simultaneously with the iron preparation, the second time at a dose of 0.08 ml / kg five days after the first injection of ferroglukin and crezacin 3 mg / kg, included in the 8-day watering schedule, starting simultaneously with the first injection of fe roglyukina.

The method allows to normalize the level of plasminogen activator inhibitor in newborn calves and piglets with anemia 3 days after completion of treatment, translating it to a level close to that for healthy animals. Subsequently, upon completion of treatment, according to the recommendations of the proposed method, it is possible to maintain the content of plasminogen activator inhibitor at an optimal level, which will allow newborn calves and piglets with anemia to significantly reduce the risk of thrombotic complications, optimize microcirculation, trophic tissue, increasing weight gain, improving the herd and speeding up obtaining a sufficient volume meat products. The inventive method is as follows.

For the diagnosis in newborn calves and piglets with anemia of a violation of the level of plasminogen activator inhibitor, it is evaluated according to the following method (Barkagan Z. S., Momot A. P. Fundamentals of the diagnosis of hemostatic disorders. M., 1999. - P.134-135).

Principle. Tissue plasminogen activator (TPA) and urokinase activate plasminogen, resulting in the formation of plasmin. These activators are neutralized by the so-called plasminogen activation inhibitors - IAP (IAP-1 and IAP-2 or PAI-1 and PAI-2). The principle of the method for determining IAP-1 is to evaluate its inactivating effect on purified urokinase in the studied plasma sample. The activity of residual urokinase is determined by its ability to convert plasminogen to plasmin. The latter is measured by its ability to cleave a chromogenic substrate at a wavelength of 405 nm.

Reagents are used: 1. Urokinase, lyophilized. It is diluted to a concentration of 5 IU / ml. 2. Plasminogen, lyophilized.

It is diluted to a concentration of 15 CTA IU / ml. 3. Chromogenic substrate: HD-Norvalyl-cyclohexylalanyl-lysyl-p-nitroanilid (HD-Nva-lys-pNA). It is diluted to a concentration of 0.6 mmol / L. 4. Solvent for the substrate. 5. The oxidizing agent is chloramine T, lyophilized, the concentration in the working solution is 1.8 g / l. 6. Tris-HCl buffer (100 mmol / L), pH 8.4. 7. Standard S1, lyophilized (0.0 U / ml IAP). 8. Standard S2, lyophilized (2.5-4.0 U / ml IAP). 9. Control plasma of a calf or pig, lyophilized (2.5-4.0 U / ml IAP).

Acetic acid, 20% solution (for the two-point evaluation method) is also required to complete the study.

The following equipment is used: spectrophotometer SF-26, SF-46 or similar. The used wavelength is 405 nm. The required width of the optical path of the cell is 1 cm.

The material for the study is a citrate platelet-poor plasma.

The diagnostic scheme for the IAP level is shown in the table.

Table. The scheme for determining the level of IAP in the examined calves and piglets. Sequence of operations Test sample Sample Form Pipette into a plastic (silicone) tube: 1. The investigated plasma / standard 0.05 ml (50 μl) 0.05 ml (50 μl) 2. Tris-HCl buffer - 0.1 ml 3. The solution of urokinase 0.1 ml - Mix and incubate at a temperature of + 37 ° C for 5 min 4. Plasminogen solution 0.2 ml 0.2 ml 5. The oxidant solution 0.2 ml 0.2 ml Mix and incubate at a temperature of + 37 ° C for 5 min 6. Chromogenic substrate 0.5 ml 0.5 ml Mix immediately and simultaneously start the stopwatch. After 5 min, acetic acid is added: 1. Acetic acid, 20% solution 0.2 ml 0.2 ml Stirred and for 2 hours in the samples determine the absorption (A) at a wavelength of 405 nm of the test sample against the sample blank.

The results are evaluated using an intralaboratory conversion factor (K _p ), which is determined using IAP standards (S1 and S2):

The activity of IAP in the studied plasma sample in% of the norm is subtracted by the formula:

IAP ACTIVITY _sample = K _p × (A _Sl - A _sample ).

Normally, the activity of IAP in blood plasma is in the range from 0.3 to 3.5 U / ml. An increase in the activity of IAP is observed in various diseases, including with anemia. A high level of IAP activity is associated with a violation of microcirculation in tissues and deterioration of trophism in them, contributing to a slowdown in the growth of calf and piglet.

If the calves and piglets with anemia increase the level of IAP in their newborns, it is necessary to prescribe treatment according to the proposed method: ferrglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, phosprenyl intramuscularly, twice in different syringes at different points, the first time 0, 08 ml / kg simultaneously with the iron preparation, the second time at a dose of 0.08 ml / kg five days after the first injection of ferroglukin and crezacin 3 mg / kg, included in the 8-day drinking schedule, starting simultaneously with the first injection of ferroglukin.

Example 1. In a newborn piglet No. 32 with anemia, a violation of erythropoiesis was detected with signs of a decrease in the level of iron in his body (serum iron 12.2 μmol / L, siderocytes 0.8%, with the amount of hemoglobin in it 79.0 g / L erythrocytes - 3.90 × 10 ¹² / L. The examination revealed an increased activity of the plasminogen activator inhibitor (4.0 units / ml).

The piglet was prescribed treatment: ferrglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, phosprenyl intramuscularly, twice in different syringes at different points, the first time 0.08 ml / kg simultaneously with the iron preparation, the second time at a dose of 0 , 08 ml / kg five days after the first injection of ferroglukin and crezacin 3 mg / kg, included in the 8-day drinking schedule, starting simultaneously with the first injection of ferroglukin.

The piglet was examined during treatment. 3 days after treatment, the anemia stopped - serum iron 26.3 μmol / L, siderocytes 7.8%, with a hemoglobin of 126.7 g / L, erythrocytes - 4.90 × 10 ¹² / L).

Assessment of plasminogen activator inhibitor activity, showing normalization of its level (2.2 u / ml). Subsequent observation of a given pig did not reveal any deviations from the norm.

Example 2. In a newborn calf No. 49 with anemia, there was a violation of erythropoiesis and signs of a decrease in the level of iron in his body (serum iron 11.5 μmol / L, siderocytes 0.9%, with a hemoglobin content of 82.0 g / L erythrocytes - 3.92 × 10 ¹² / L. The examination revealed an increased activity of the plasminogen activator inhibitor (4.0 units / ml).

The calf was prescribed treatment: ferrglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, phosprenyl intramuscularly, twice in different syringes at different points, the first time 0.08 ml / kg simultaneously with the iron preparation, the second time at a dose of 0 , 08 ml / kg five days after the first injection of ferroglukin and crezacin 3 mg / kg, included in the 8-day drinking schedule, starting simultaneously with the first injection of ferroglukin.

The calf was examined during treatment. 3 days after treatment, the anemia stopped - serum iron 27.1 μmol / L, siderocytes 8.0%, with a hemoglobin content of 125.9 g / L, erythrocytes - 5.00 × 10 ¹² / L).

Assessment of plasminogen activator inhibitor activity, showing normalization of its level (2.0 units / ml). Subsequently, observation of this calf did not reveal any deviations from the norm.

Using the proposed method for normalizing the level of plasminogen activator inhibitor in veterinary medicine will help to avoid vascular complications in newborn calves and piglets with anemia, deterioration of trophism in tissues due to inefficient microcirculation, reduce mortality, increase and improve the number of cows and pigs.

Claims (1)

A method of normalizing the level of plasminogen activator inhibitor in case of anemia in newborn calves and piglets with anemia, including the use of ferroglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, fosprenil intramuscularly, twice in different syringes at different points, the first time 0.08 ml / kg simultaneously with the iron preparation, the second time at a dose of 0.08 ml / kg five days after the first injection of ferroglukin and crezacin 3 mg / kg, included in the 8-day drinking schedule, starting simultaneously with the first injection of ferroglukin.