Method for predicting pregnancy in cows

FIELD: animal science, cattle science.

SUBSTANCE: the innovation deals with testing blood serum for the content of ceruloplasmin and at the value of its activity being above 0.095 U optic density it is possible to diagnose the pregnancy. The technique suggested is accurate, enables to diagnose the pregnancy in cows at early stage and, also, it enables to increase efficiency, decrease labor capacity and avoid the usage of expensive devices and reagents.

EFFECT: higher accuracy of diagnostics.

1 tbl

 

The invention relates to livestock, in particular cattle, and can be used for early diagnosis of pregnancy in cows.

Currently, methods for diagnosis of pregnancy are divided into two groups:

1) methods of clinical diagnosis;

2) methods of laboratory diagnostics.

Laboratory methods based on the detection of specific changes in hormonal function of ovaries or placenta metabolism of the mother and into her body metabolic products of the fruit. From numerous laboratory methods for diagnosis of pregnancy is the most accurate hormonal.

Laboratory methods the most frequently used method for the determination of progesterone in the blood. Radioimmunoassay method for the determination of the concentration of progesterone is based on the fact that luteum produces progesterone pregnant during the whole period of pregnancy. Its concentration with 5-7-th day after insemination increases, reaching a peak at 12-17 day, and if pregnancy does not occur, then for 1-3 days prior to the regular sexual cycle, the level of progesterone drops sharply from 4.9 ng/ml and more than 1.7 ng/ml (Means VA "Biotechnology of reproduction in cattle. Tutorial, Nalchik, 2004, str-275).

The disadvantages of all laboratory methods are laborious, complexity, small Cycling and the use of expensive instruments and reagents.

The objective of the proposed method is to increase productivity, reduce labor intensity and thoroughness, and rejection of the use of expensive instruments and reagents.

The problem is solved due to the fact that in the known method for the diagnosis of pregnancy in cows, which consists in the study of serum level of progesterone, according to the invention examine the blood on the content of ceruloplasmin and when it is more 0,095 units of optical density diagnose pregnancy.

Pregnancy is characterized by a high level of free-radical oxidation.

During pregnancy there is also a parallel increase in the activity of the antioxidant system. The growth rate of antioxidant activity is the activity of one of the antioxidant ceruloplasmin.

Ceruloplasmin is a protein with an electrophoretic mobility of alpha2- globulin, is mesotelioma oxidase (CF)involved in the acute phase of inflammation and stress. Discovered antioxidant properties of ceruloplasmin, is synthesized by many tissues.

Determination of the activity of ceruloplasmin were rapid method for Aviano (1981).

The method is based on the ability of ceruloplasmin to show catalytic activity as polyphenoloxidase. As a result of okalinoobrazovaniya formed substances orange the colour intensity is proportional to the concentration of ceruloplasmin.

To clarify the essence of the proposed method we present methods for determining the activity of ceruloplasmin in the serum.

Required reagents:

solution No. 1: 0,4M sodium acetate buffer (pH of 5.6). To prepare the solution take 1 ml of 0.4 M CH3COOH and 7.2 ml of 0.4 M CH3COONa.

solution # 2: the 1%aqueous solution of paraphenylenediamine hydrochloric acid. Take 1 g of paraphenylenediamine hydrochloric acid and dissolved in 99 ml of H2O.

solution # 3: a 25%solution of caustic soda (NaOH). Take 25 g of NaOH and dissolved in 75 ml of N2O.

The method is as follows.

In tubes placed 0.1 ml of serum, add 1 ml of 0.4 M ateenage buffer (pH 5.6) and 0.5 ml of 1% aqueous solution of paraphenylenediamine hydrochloric acid. Mixed and placed in a water bath at 60°C.

After 10 min the reaction is stopped by adding 3.5 ml of chilled 25%caustic soda solution. The obtained orange liquid colorimetrate at 440 nm.

In the control samples contained all of the above components, but the sodium hydroxide solution is added to the tubes before placing them in a water bath.

The activity of ceruloplasmin was expressed in units of optical density (conventional units).

In our experiments we used 15 cows of black-motley breed in age is e 3-5 years with a live weight of 350-450 kg and the average milk yield for 305 days of lactation 3110 kg of milk.

The research results presented in the table.

The activity of ceruloplasmin in the serum of pregnant and non-pregnant cows
Non-pregnant women (not pregnant) animalsPregnant (pregnancy 24 days)Pregnant (pregnancy 48 days)
no animalThe activity of ceruloplasmin in the unit from. squareThe activity of ceruloplasmin in the unit from. squareThe activity of ceruloplasmin in the unit from. square
10,070,100,120
20,0730,110,125
30,0740,0950,130
40,0720,0950,120
50,070,100,130

Thus, as clearly seen from the table, when the value of ceruloplasmin more 0,095 units of optical density diagnose pregnancy (pregnancy) in cows at an early stage with high precision and objectivity.

Method for diagnosis of pregnancy in cows, which consists in the study of blood serum, wherein the serum is examined for content is ceruloplasmin, and when the value of its activity above 0,095% opt. pl. diagnose pregnancy.



 

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