RU2403022C1 - Method of protein c activity optimisation in anemia of newborn calves and piglets - Google Patents

Abstract

FIELD: medicine, veterinary science.

SUBSTANCE: invention refers to biology. 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 0.07 ml/kg combined with an iron preparation, and the second time in dosage 0.07 ml/kg six days after the first injection of ferroglucin and cesacine 4 mg/kg that is included in the feed regimen for 10 days.

EFFECT: method allows optimising protein C activity in newborn calves and piglets with anaemia 2 days after the termination of treatment, maintaining its optimum condition, reducing risk of thrombotic complications, improving a herd, increasing weight gain, reducing mortality and providing quality and amount of produced meat.

1 tbl, 2 ex

Description

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

Analogues of the proposed method for optimizing protein C activity 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 in case of anemia (Internal non-communicable diseases of farm animals. / Under the editorship of V. M. Danilevsky. M.: Agropromizdat, 1991. - 576 p.).

In modern veterinary medicine, phosprenyl is used as an effective stimulant of the vital activity of a growing organism. Phosprenyl is a drug based on polyprenol phosphates obtained from needles (Deeva A.V., Salakhova Z.A., Lobova T.P. et al. 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) - an effective stimulator of living organisms that increases the survival of animals in adverse and extreme conditions, stimulating or not malizuya life processes (Baymatov VP, Latypov MM Influence krezatcina on the state bird. Veterinary 2006, №11, s.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 optimize the activity of protein C.

The aim of the invention is to increase the efficiency of correction of disorders of protein C activity in newborn calves and piglets with anemia.

Protein C is a vitamin K-dependent glycoprotein synthesized in hepatocytes that circulates in the blood as a proenzyme. Under the action of the thrombin-thrombomodulin complex (or an activator from snake venom), protein C activates and acts as an anticoagulant, causing proteolysis of factors Va and VIIIa in the presence of its cofactor - protein S and phospholipids. With thrombophilia in newborn calves and piglets, including in case of anemia, disorders in the hemostatic system may be due to protein C deficiency.

The essence of the proposed method lies in the fact that to optimize the activity of protein C newborn calves and piglets with anemia are prescribed ferroglukin 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 , 07 ml / kg simultaneously with the iron preparation, a second time at a dose of 0.07 ml / kg six days after the first injection of ferroglukin, and crezacin 4 mg / kg when it is included in the 10-day rehydration schedule.

The method allows to optimize the activity of protein C in newborn calves and piglets with anemia 2 days after the end of treatment, translating it to a level close to that for healthy animals. Subject to the recommendations of the proposed method, it is possible to maintain protein C activity in optimal condition in the future, which will allow newborn calves and piglets with anemia to significantly reduce the risk of thrombotic complications, improve the herd, increase the weight gain, reduce the case and increase the quality and volume of meat products.

The inventive method is as follows.

For the diagnosis of newborn calves and piglets with anemia of a disturbance in the level of protein C in the blood, it is evaluated according to the following procedure (Barkagan ZS, Momot AP, Fundamentals of the diagnosis of hemostatic disorders. M, 1999. - P.118-120).

Protein C in the samples of the studied plasma is activated by a specific activator from the muzzle poison - Agkistrodon contortrix. When activated with this protein C causes proteolysis of factors Va and VIIIa in a protein C deficient plasma. The latter leads to an increase in clotting time, which is used to evaluate the activity of protein C in the test sample. Protein C activity, expressed as a percentage of normal, is found from a previously constructed calibration curve.

Reagents are used: 1. Protein activator, lyophilized, diluted with 3 ml of distilled water. 2. Plasma deficient in protein C, lyophilized, is diluted in 1 ml of distilled water. 3. Neothromtin, lyophilized - an analogue of the kaolin-cephalic reagent, containing instead of kaolin ellagic acid and phospholipids, 3 ml of distilled water is diluted.

In addition, a medial buffer (pH 7.6), used to dilute samples of the test and standard plasma, and a solution of calcium chloride (0.025 M) are required. To construct a calibration curve, a standard branded or control normal plasma with known protein C activity or a pooled plasma (taken from at least 10 healthy calves or piglets, respectively) is needed, the protein C activity in which is taken as 100%.

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

Before the analysis, the studied plasma is diluted with a medial buffer 10 times.

0.1 ml of diluted (1:10) plasma, 0.1 ml of protein C deficient in protein C, 0.1 ml of activator are preheated at the same temperature in a test tube (or coagulometer cuvette), heated to + 37 ° C. protein C and 0.1 ml of the kaolin-mullet analogue reagent. The mixture is kept in a water bath at

+ 37 ° C for 4 minutes Then, 0.1 ml of a solution of calcium chloride, preheated to 37 ° C, is added to the mixture, a stopwatch or a coagulometer timer is turned on, and the clotting time is recorded.

Results are measured by a calibration curve.

Construction of a calibration curve. Prepare a series of dilutions of the control plasma pool or standard branded plasma using a medial buffer (table).

Scheme for preparing plasma dilutions for constructing a calibration curve when determining the activity of protein C No. Protein C,% Breeding Getting dilutions one 200 1: 5 0.2 ml of plasma + 0.8 ml of buffer 2 one hundred 1:10 0.1 ml of plasma + 0.9 ml of buffer 3 fifty 1:20 0.1 ml of plasma + 1.9 ml of buffer four twenty 1:50 0.1 ml of dilution No. 1 + 0.9 ml of buffer 5 10 1: 100 0.1 ml of dilution No. 2 + 0.9 ml of buffer

With each of the samples of the diluted plasma, the clotting time is determined by the above procedure.

When constructing a calibration curve on the logarithmic axis of ordinates, coagulation time in seconds is postponed, on the logarithmic axis of abscissa, protein C activity in% in accordance with the dilutions indicated in the table.

Normally, protein C activity is 70-140%.

When establishing a decrease in protein C activity, newborn calves and piglets with anemia should be prescribed the following treatment complex.

The essence of the proposed method lies in the fact that to optimize the activity of protein C newborn calves and piglets with anemia are prescribed ferroglukin 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 , 07 ml / kg simultaneously with the iron preparation, a second time at a dose of 0.07 ml / kg six days after the first injection of ferroglukin, and crezacin 4 mg / kg when it is included in the 10-day rehydration schedule.

Example 1. In a newborn calf with anemia No. 07, examination revealed a decrease in the activity of protein C - 55%. So, the amount of hemoglobin in him was 81.0 g / l, erythrocytes 4.01 × 10 ¹² / l, siderocytes 1.0% with a serum iron level of 12.2 μmol / l with an increase in the total iron binding capacity of blood serum to 67, 0 μmol / L.

The calf was prescribed ferroglukin 150 mg intramuscularly, twice with an interval of 10 days, phosprenyl intramuscularly, twice in different syringes at different points, the first time 0.07 ml / kg simultaneously with the iron preparation, the second time at a dose of 0.07 ml / kg six days after the first injection of ferroglukin, and crezacin 4 mg / kg when it is included in the 10-day rehydration schedule.

2 days after the end of treatment, the calf improved its red blood counts. So, the amount of hemoglobin in it increased to 126.0 g / l, erythrocytes - to 4.92 × 10 ¹² / l, siderocytes to 8.0% with an increase in serum iron to 26.6 μmol / l and a decrease in the total iron binding capacity blood serum up to 32.4 μmol / l.

An increase in the level of protein C activity (84%) to the norm was obtained, which subsequently remained at the achieved level.

Example 2. In a newborn piglet with anemia No. 17, examination revealed a decrease in protein C activity - 59%.

In this piglet there was a violation of erythropoiesis with signs of a decrease in the level of iron in his body, accompanied by a change in blood counts. So, the amount of hemoglobin in him was 79.0 g / l, erythrocytes 3.96 × 10 ¹² / l, siderocytes 1.1% with a serum iron level of 12.0 μmol / l with an increase in the total iron binding capacity of blood serum to 68, 2 μmol / L.

The animal was prescribed 150 mg ferroglukin intramuscularly, twice with an interval of 10 days, fosprenyl intramuscularly, twice in different syringes at different points, the first time 0.07 ml / kg simultaneously with the iron preparation, the second time at a dose of 0.07 ml / kg six days after the first injection of ferroglukin, and crezacin 4 mg / kg when it is included in the 10-day rehydration schedule.

2 days after the end of treatment, the red blood returned to normal in the piglet. So, the amount of hemoglobin in it increased to 120.7 g / l, erythrocytes - up to 5.36 × 10 ¹² / l, siderocytes up to 8.3% with an increase in serum iron to 28.3 μmol / l and a decrease in the total iron binding capacity blood serum up to 32.0 μmol / l.

Complete normalization of protein C activity (109%) was achieved, which later did not experience negative dynamics.

Using the proposed method for optimizing protein C activity in veterinary medicine and biology will help to avoid many vascular complications in newborn calves and piglets with anemia, improve the herd, reduce mortality, increase weight gain, increase the quality and volume of meat products and get healthy from these calves and piglets progeny.

Claims (1)

A method for optimizing the activity of protein C in newborn calves and piglets with anemia by applying ferroglukin 150 mg intramuscularly, twice with an interval of 10 days, fosprenil intramuscularly, twice in different syringes at different points, the first time 0.07 ml / kg at a time with an iron preparation, a second time at a dose of 0.07 ml / kg six days after the first injection of ferroglukin and crezacin 4 mg / kg when it is included in the 10-day watering schedule.