RU2262699C2 - Preparation applied for predicting subclinical mastitis in cattle - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: the suggested preparation consists of sulfonol and an indicator as neutral red, methylene blue and it additionally contains caustic potash at the following ratio of components, weight%: technical sulfonol 4.75-4.77, neutral red 0.001-0.003, methylene blue 0.001-0.003, caustic potash 0.09-0.11, water - the rest. This preparation enables to predict subclinical mastitis in cattle at earlier stage of disease.

EFFECT: higher accuracy and efficiency of diagnostics.

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Description

The invention relates to veterinary medicine and can be used to diagnose subclinical mastitis in cows.

A diagnostic tool for subclinical mastitis is known by determining lysozyme in milk. It is based on the ability of milk lysozyme to inhibit the growth of the test microbe (Staphylococcus aureus strain VM). According to the growth inhibition zone, the titer of lysozyme in milk and breast health are judged (3).

Using this diagnostic tool in a farm environment is practically difficult to implement, since it requires a specially equipped laboratory and certain skills in conducting microbiological studies, a microorganism culture is required.

Known means of determining the amount of chloride in milk as an indicator of the presence of subclinical mastitis. Healthy cow's milk contains 0.08-0.14% chloride. With mastitis, their number in the contents of the udder increases. 5 ml of a 1.5% solution of silver nitrate and 2 drops of a 10% aqueous solution of potassium chromate are added to the tubes, and 1 ml of the test milk is added. The contents of the tubes are mixed. If the amount of chloride in the test milk exceeds 0.14%, the mixture turns yellow (2).

The disadvantage of this tool is that the concentration of chlorine ions necessary for determining is not always created in milk.

Known diagnostic tool for subclinical mastitis, based on the determination of chlorine-sugar number (ratio of chlorides and lactose). Lactose (milk sugar) is determined by the iodometric method (5).

The disadvantage of this tool is that this chlorine-sugar number can change with other diseases, including ketosis, and thereby lead to distorted results.

A diagnostic tool for subclinical mastitis is known by measuring the pH of milk. If the milk of a healthy animal has a pH of 6.3–6.9 on average, then in pathological conditions the reaction of the medium shifts to the alkaline side — 7.0 and higher. For this, 2-4 drops of the test milk are applied to a white porcelain plate and 1-2 drops of a 0.2% alcohol (65 degree alcohol) bromothymol blue solution are added to them. Dyeing milk in yellow or greenish yellow is taken as an indicator of normal milk; blue-green or green-yellow staining of the sample is a sign of subclinical mastitis (4).

The disadvantage of this diagnostic tool is that the secretion of the mammary gland has a high buffer capacity and neutralizes hydrogen ions, preventing the indication of a changed pH.

Known diagnostic tools for subclinical mastitis, based on the detection of blood pigments in milk. For this, 5 ml of a 3% solution of hydrogen peroxide and 2 ml of a saturated solution of benzidine in glacial acetic acid are added to the test tube. The contents of the tube are thoroughly shaken and then the test milk (2-10 drops) is added dropwise to the tube. By the number of drops of milk introduced into the test tube, at which a positive reaction appeared, we can tentatively judge the number of blood pigments in the milk. With a positive reaction, the contents of the tube initially become greenish, and after 5-10 minutes it becomes dark blue. In case of a negative reaction, the contents are light; a whitish flocculent precipitate appears at the bottom of the tube (1, 4).

The disadvantage of this tool is that in the initial stages of subclinical mastitis, the reaction to blood pigments is not always a reliable sign of a developing disease, while for some physiological and pathological conditions of the udder, not associated with mastitis, they will be positive.

Known diagnostic tools for subclinical mastitis, based on the combined use of surfactants and indicators. Based on this principle, a drug such as dimastin is used, which we have chosen as a prototype. Diagnostics by these agents is based on the formation of a clot from the nuclei of somatic cells, the amount of which increases in milk with mastitis, and a change in its color (a change in pH and, as a result, a change in the color of the indicator). When diagnosing mastitis with a 5% solution of dimastine, the reaction is taken into account according to the following indicators: negative reaction (no mastitis) - when mixing 1 ml of a 5% solution of dimastine with 1 ml of milk a homogeneous liquid is formed, the color is orange, orange-red. Doubtful reaction - traces of jelly formation, color - red. A positive reaction is a clearly visible clot that is half or completely thrown out of the hole with a stick with stirring, color: scarlet, crimson, raspberry (3).

The advantage of these tools is that they can be used in any household without taking the cost of expensive equipment.

The disadvantages of these drugs are that, firstly, their composition does not take into account the high buffer capacity of milk and the indicator begins to change its color with a significant shift in the reaction of the medium; secondly, the indicators included in their composition, for example phenol red in dimastine, begin to change color at a pH value of 7.2 or higher, while in subclinical mastitis, a change in pH, indicating the onset of the disease, occurs from 7, 0 and above.

The aim of the invention is a diagnostic tool for subclinical mastitis in cows, allowing you to register this disease at an earlier stage.

This goal is achieved by the fact that when developing a diagnostic tool for subclinical mastitis, the features of the secretion of the udder are used for this disease to contain an increased number of somatic cells and have a hydrogen index above 7.0. For this, the product contains (wt.%): Technical sulfonol 4.75-4.77, neutral red 0.001-0.003, methylene blue 0.001-0.003, caustic potassium 0.09-0.11 to reduce the buffer capacity of milk.

To indicate an increased number of somatic cells in milk, they use their property to form a jelly-like clot upon contact with a surfactant. Surfactant, destroying the nuclei of somatic cells, forms complexes with their proteins and displaces DNA from the bound (with the protein) to the free polymer state, in which it is viscous due to the peculiar structure of the macromolecule.

Technical surfactants are used as surfactants. A solution is prepared from it, 100 g of which contains 5 g of powder. The solution is used after 3 days of exposure.

To register the change in pH, we chose an indicator consisting of equal parts of a 0.1% aqueous solution of neutral red and a 0.1% solution of methylene blue. This mixture has a clear color transition with a change in pH. In an acidic environment, it has a blue-violet color, and at pH = 7.0 and above it turns into green.

The method of preparation of the reagent (the reagent is hereinafter referred to as the Trinity Mastitis Test (TMT)): the indicator is mixed with a solution of sulfonol 1:20, caustic potassium 0.1 g per 100 g of solution is added.

A comparable analysis of the claimed and known diagnostic tools allows you to identify signs that distinguish the proposed technical solution from the prototype: the synergistic use of caustic potassium in the mixture, the presence of which reduces the high buffer capacity of milk, and an indicator that records an earlier change in pH due to this.

Thus, the claimed diagnostic tool subclinical mastitis in cattle meets the criterion of "novelty."

The authors propose to use the reagent for the diagnosis of subclinical mastitis as follows: pour 1 ml of milk of the last portion (alveolar milk) from each quarter of the wells of the control plate. Then mix the test milk with 1 ml of reagent in the well of the milk control plate.

The reaction should be taken into account after 4 seconds according to the degree of jelly (clot) formed and the color change of the indicator.

The reaction is considered:

- negative (no mastitis) if there is a uniform violet liquid in the well;

- it is doubtful if there are traces of jelly formation in the hole, which, when mixed with a glass or wooden stick, stretches behind it in the form of thin threads, the color of the mixture is purple;

- positive, if a thick clot forms, which, when stirred, is easily thrown out of the hole with a glass rod, the color of the mixture is purple, or if a thick clot forms, and the mixture turns greenish or bright green.

Example:

- to implement the proposed tool and method for the diagnosis of subclinical mastitis, a scientific and production experiment was conducted.

Under the conditions of Schelkunskoye OJSC in the Sysert district of the Sverdlovsk region, 100 cows of black-motley breed with a dimastin test were examined, which revealed 8 animals with subclinical mastitis, or 8%. Then the same cows were examined for the presence of subclinical mastitis with TMT reagent. Studies have fully confirmed the results of the dimastin test. Additionally, 7 goals were identified. The total number of animals with subclinical mastitis was 15 animals or 15%.

1. Results of studies on subclinical mastitis

Number of animals, goal Revealed sick cows with dimastine breakdown, goal Revealed sick cows breakdown "TMT", goal Additionally identified patients with a breakdown of "TMT", goal 100 8 fifteen 7

Milk from sick cows was delivered to the veterinary laboratory, where E. coli, Str. uberis, St. epidermidis - causative agents of subclinical mastitis.

Thus, for the diagnosis of subclinical mastitis in cattle, we can use the tool we have proposed to increase the sensitivity of determination, reduce the cost of preventive and curative work, and increase labor productivity.

The drug is a foaming green liquid. Shelf life is 1 year.

List of references

1. Alekseev, I.A., Mirolyubov, M.G. Experience in combating mastitis in dairy complexes. - Cheboksary: Chuvash book publishing house, 1984. - 48 p.

2. Valyushkin, K.D., Medvedev, G.F. Obstetrics and Biotechnology of Animal Reproduction: Textbook, 2nd ed., Rev. and add. - Мn .: Urajay, 2001 .-- 869 p.

3. Ivashura, A.I. The system of measures to combat mastitis of cows. - M :. Rosagropromizdat, 1991 .-- 240 p.

4. Ilyina A.I., Pospelov A.I. Diseases of the udder. L.-M., Selkhozgiz, 1961, 152 p.

5. Kartashova, V.M., Ivashur, A.I. Mastitis of cows. - M.: Agropromizdat, 1988 .-- 256 p.

Claims (1)

A tool for the diagnosis of subclinical mastitis in cattle containing sulfonol and an indicator, characterized in that neutral red, methylene blue is used as an indicator and it additionally contains caustic potassium in the following ratio of components, wt.%: Sulfonol technical 4.75-4.77 Neutral red 0.001-0.003 Methylene blue 0.001-0.003 Potassium hydroxide 0.09-0.11 Water  Rest