RU2399374C1 - Method of plasma plasminogen activator inhibitor concentration optimisation in newborn piglets with anemia - Google Patents

Abstract

FIELD: medicine, veterinary science.

SUBSTANCE: invention refers to veterinary science and biology, namely to haematology. The method involves double intramuscular introduction of ferroglucin 150 mg (2 ml), every 10 days. Besides polyzone 4 mg/kg and cresacin 4 mg/kg are included in the feed regimen for 8 days.

EFFECT: method allows avoiding vascular complications in newborn piglets with anaemia, tissue trophic reduction caused by noneffective microcirculation, reducing mortality, increasing and improving swine population.

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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 piglets with anemia do not exist.

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

In modern veterinary medicine, polyisone, a phosphoric acid salt of 2-amino-4-methylthio- (S-oxo-S-amino) -butyric acid, is a weakly soluble white to cream-colored powder (Ovchinnikov) as effective stimulants of the vital activity of a growing organism in modern veterinary medicine. A. et al. Polizon - growth promoter // Poultry, 2006. - No. 12. - p.14-15) and crezacin - a derivative of aroxyalkylcarboxylic acids. It is a white crystalline powder, readily soluble in water, with a faint specific smell, sweetish-bitter taste. It exhibits an effective stimulating effect on living organisms, increasing the survival of humans and animals in adverse and extreme conditions, stimulating or normalizing life processes (Baymatov V.N., Latypov M.M.The effect of krezacin on the state of birds. Veterinary Medicine, 2006, No. 11, p. 49-51).

However, never before has a treatment complex consisting of ferroglukin, polysone and crezacin been used in newborn 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 piglets with anemia.

The essence of the proposed method lies in the fact that to optimize the level of plasminogen activator inhibitor, newborn piglets with anemia are prescribed ferroglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, polyzone 4 mg / kg and crezacin 4 mg / kg included in the scheme drinking for 8 days.

The method allows to normalize the level of plasminogen activator inhibitor in newborn 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 piglets with anemia to significantly reduce the risk of thrombotic complications, optimize microcirculation, trophic tissue, increasing weight gain, improving the herd and accelerating the receipt of a sufficient volume of meat products .

The inventive method is as follows.

For diagnosis in newborn piglets with anemia of a violation of the level of plasminogen activator inhibitor, it is assessed according to the following method (Barkagan Z.S., Momot A.P. Basics for the diagnosis of hemostasis 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. The control plasma of the 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 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 - Stir and incubate at + 37 ° C for 5 min 4. Plasminogen solution 0.2 ml 0.2 ml 5. The oxidant solution 0.2 ml 0.2 ml Stir and incubate at + 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 internal laboratory 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 _S1 -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 piglet growth.

With an increase in newborn piglets with anemia, the level of IAP requires the appointment of treatment according to the proposed method.

To normalize the level of plasminogen inhibitor, newborn piglets with anemia are prescribed treatment according to the proposed method: ferroglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, polyzone 4 mg / kg and crezacin 4 mg / kg included in the 8-day rehydration schedule .

Example. 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 12.0 μmol / L, siderocytes 0.8%, with a hemoglobin content of 78.0 g / L, erythrocytes - 3.95 × 10 ¹² / L). An examination revealed an increased activity of the plasminogen activator inhibitor (4.0 u / ml).

The treatment was prescribed for the piglet: ferroglukin 150 mg (2 ml) intramuscularly, twice with an interval of 10 days, polyzone 4 mg / kg and crezacin 4 mg / kg included in the 8-day watering schedule.

The piglet was examined during treatment. 3 days after treatment, the anemia stopped - serum iron 22.6 μmol / L, siderocytes 7.8%, with the amount of hemoglobin he had 123.8 g / L, red blood cells - 4.95 × 10 ¹² / L).

Assessment of plasminogen activator inhibitor activity, showing normalization of its level (2.2 u / ml). Subsequently, observation of this pig 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 piglets with anemia, deterioration of trophism in tissues due to inefficient microcirculation, reduce mortality, increase and improve the number of pigs.

Claims (1)

A method for optimizing the plasma plasminogen activator inhibitor content in newborn piglets with anemia, including the use of 150 mg (2 ml) of ferroglukin intramuscularly, twice with an interval of 10 days, polyzone 4 mg / kg and crezacin 4 mg / kg included in 8 days.