RU2419093C1 - Method to forecast development of toxic form of dyspepsia in newborn calves with initial symptoms of dyspepsia - Google Patents

Abstract

FIELD: veterinary science. ^ SUBSTANCE: in newborn calves with initial dyspepsia symptoms blood samples are taken to identify aggregative activity of platelets using solutions of ADP, collagen, ristomycin and adrenalin as inducers by mixing of 0.1 ml of plasma rich in platelets and standardised by platelets quantity on a slide plate with 0.1 ml of one of aggregating agents and identifying time of platelet aggregates appearance in the form of whitish grain visible with the naked eye. The reference is the average result of identified aggregation of platelets in 10 healthy newborn calves. If platelets aggregation acceleration over 7% compared to the reference is detected in newborn calves with initial dyspepsia symptoms, development of toxic dyspepsia is forecasted. Early forecasting of toxic dyspepsia development in newborn calves has signification practical value, since it makes it possible to more thoroughly treat animals. ^ EFFECT: reduced loss of calves sick with dyspepsia and improved economic efficiency of stock raising. ^ 1 tbl, 2 ex

Description

The invention relates to veterinary medicine, namely to the diagnosis of animal diseases.

At present, it has been established that in some pathological conditions of animals, changes in platelet aggregation activity are observed.

There are various methods for determining the functional ability of platelets based on the assessment of spontaneous or induced (using various agonists) platelet aggregation (Barkagan ZS, Momot AP Diagnosis and controlled therapy of hemostasis disorders. - M.: Newdiamed-AO) , 2001 .-- 296 p.).

A known method for determining platelet aggregation activity "on glass" (Shitikova A.S. Visual micromethod for determining platelet aggregation activity with various aggregating agents / A.S. Shitikova // Laboratory work. - 1984. - No. 4. - P.215-220 )

However, all these methods are used only to assess the functional activity of platelets in various pathologies, and are not used to predict the development of a toxic form of dyspepsia.

The purpose of the invention is the prediction of newborn calves with initial signs of dyspepsia of the development of a toxic form of dyspepsia.

Early prediction of the development of toxic dyspepsia in newborn calves is of great practical importance, because It allows a more thorough approach to therapy and timely preventive treatment of diseased animals, which will help reduce the incidence of dyspepsia in newborn calves and increase the economic efficiency of animal husbandry.

The essence of the proposed method is as follows. Blood samples are taken from newborn calves with initial signs of dyspepsia and platelet aggregation activity is determined using ADP, collagen, ristomycin and adrenaline solutions as inducers by mixing on a glass slide 0.1 ml of platelet-rich plasma standardized for platelet count and 0.1 ml of one from aggregating agents and determining the time of manifestation of platelet aggregates in the form of whitish grains visible to the naked eye. As a control, the average result of determining induced platelet aggregation in 10 clinically healthy newborn calves is used. If newborn calves with initial signs of dyspepsia are found to have accelerated platelet aggregation of more than 7% of the control, toxic dyspepsia can be predicted.

The method is as follows: in newborn calves with initial signs of dyspepsia, blood samples are taken from morning until feeding in a plastic dish. To prevent spontaneous aggregation, stabilization of the obtained blood with a 3.8% sodium citrate solution in a ratio of 1: 4 is used. The induced platelet aggregation activity is determined using solutions of ADP (dilution of the mother liquor 1:10), collagen (concentration 20 mg / ml), ristomycin (concentration 15 mg / ml) and adrenaline (dilution of the mother liquor 1:50) as inducers. For this purpose, platelet-rich plasma and platelet-poor plasma are obtained from stabilized blood. The platelet-rich plasma used is obtained by centrifuging the obtained stabilized blood at 1000 rpm for 6 minutes; and platelet-poor plasma after centrifugation of the remaining platelet-rich plasma at 3000 rpm for 20 minutes after centrifugation. Platelet-rich plasma is standardized for platelet count by diluting, if necessary, with an autologous sample of platelet-poor plasma (up to 380-400,000 platelets / μl). The volumes of mixed plasma rich and poor in platelets are determined by the formula (1):

where V _ppp is the volume of platelet-poor plasma,

V _prp is the volume of platelet-rich plasma,

N is the counted platelet concentration in the original platelet-rich plasma (cells / μl).

Then, 0.1 ml of the investigated platelet-rich plasma and 0.1 ml of one of the aggregating agents are applied with different pipettes to a fat-free dry glass slide. After mixing the droplets with a glass rod, the stopwatch is immediately turned on and the time of manifestation of the so-called snowstorm phenomenon is established — the moment when the platelet aggregates reach such sizes that they can be seen with the naked eye in the form of whitish grains (it is necessary to use the illumination for a microscope like OI-19 ) The snow storm phenomenon in healthy individuals occurs at the same time, and when the aggregation function is strengthened, it appears earlier than normal, and when it weakens, later. The reaction is repeated 2 times with the same aggregating agent. The result is taken into account in seconds. The result is the average of two definitions.

As a control, the average result of determination by the above-described method of induced platelet aggregation in 10 clinically healthy newborn calves of the same age group is used.

If newborn calves with initial signs of dyspepsia are detected, accelerated platelet aggregation of more than 7% of the control can predict the development of a toxic form of dyspepsia.

Example 1. According to the principle of analogues, taking into account the origin, age and live weight at birth, 2 groups of newborn calves were formed. The conditions for keeping, feeding and caring for animals were the same. The first group consisted of 22 clinically healthy newborn calves. The second group was made up of 17 calves with initial signs of dyspepsia, who were treated according to the scheme adopted at the farm (tylosin-50 0.2 ml / kg intramuscularly 1 time per day before recovery; decoction of horse sorrel on isotonic sodium chloride solution in the morning and evening 0.5 l each). During the experiment, the second group was divided into two subgroups: patients with a simple (n = 10) and patients with a toxic form of dyspepsia (n = 7) calves. The criterion for this separation was the difference in the clinical picture of the disease. For calves with simple dyspepsia, frequent defecation with the release of liquefied feces was characteristic, the general condition of the sick calves remained satisfactory. In calves suffering from the toxic form of dyspepsia, with the full development of the picture of the disease, profuse diarrhea was observed, depression was observed, adynamia, emaciation, ruffle of the coat, dry nasal speculum.

In patients with calf dyspepsia (n = 17), platelet aggregation was determined at the beginning of the disease (with the first signs of dyspepsia), at the height of the disease (on day 2-3 of dyspepsia), and at the stage of recovery.

The results of determining induced platelet aggregation in clinically healthy and dyspeptic calves are presented in table 1. As can be seen from table 1, in patients with dyspepsia of newborn calves, in the beginning of simple dyspepsia, in comparison with the average data of clinically healthy newborn calves, there was an acceleration of induced aggregation by 5.51% when using collagen, by 6.26% when using ADP, by 5.95% when using ristomycin and by 5.80% when using adrenaline. With toxic dyspepsia at the onset of the disease, the aggregation results were ahead of the average data of healthy newborn calves by 7.72% with collagen, 8.39% with ADP, 7.82% with ristomycin and 8.09% with adrenaline.

Table 1. Induced Platelet Aggregation (AT) of clinically healthy and dyspeptic neonatal calves Inductors Mean AT of healthy newborn calves (n = 22) Simple dyspepsia (n = 10) Toxic dyspepsia Disease onset The height of the disease Stage of recovery The onset of the disease (n = 7) The height of the disease (n = 7) Stage of recovery (n = 5) Collagen s 29.92 ± 1.16 28.27 ± 0.31 * 25.30 ± 0.07 * 29.29 ± 0.07 * 27.61 ± 0.36 * 17.59 ± 0.12 * 25.47 ± 0.11 * ADP, s 38.97 ± 1.61 36.53 ± 0.39 * 32.94 ± 0.07 * 37.06 ± 0.07 * 35.70 ± 0.40 * 23.03 ± 0.06 * 34.13 ± 0.14 * Ristomycin, with 41.02 ± 1.61 38.58 ± 0.37 * 36.17 ± 0.06 * 39.49 ± 0.08 * 37.81 ± 0.39 * 24.11 ± 0.10 * 36.33 ± 0.18 * Adrenaline, with 97.05 ± 4.01 91.42 ± 1.08 * 75.53 ± 0.07 * 94.07 ± 0.06 * 89.20 ± 1.24 * 57.07 ± 0.10 * 87.67 ± 0.48 * * -p≤0.05 compared to the average AT of healthy newborn calves

Accordingly, even when the first signs of dyspepsia are manifested, when it is still impossible to diagnose the form of the disease, platelet aggregation occurs earlier than 7% in those newborn calves who are subsequently ill with the toxic form of dyspepsia. Thus, the determination of induced platelet aggregation in newborn calves by the proposed method can be used as a relatively simple and affordable method for predicting the development of a toxic form of dyspepsia in newborn calves with initial signs of dyspepsia.

Example 2. The newborn calf No. 122 revealed the initial signs of dyspepsia: rapid bowel movements with the release of liquefied feces. To assess the risk of developing a toxic form of dyspepsia in an animal, a blood sample was taken, platelet aggregation activity was determined using collagen as an inducer. This indicator in a sick calf is 27.25 s. As a control, we used the average result of determining platelet aggregation activity in 10 clinically healthy newborn calves (30.05 s). The acceleration of platelet aggregation in a diseased calf is 2.80 s or 9.32%. Accordingly, in this animal, the development of a toxic form of dyspepsia can be predicted. The calf was prescribed adequate therapy: 0.1 ml of a 0.1% solution of the Biopag-D preparation per 1 kg of calf live weight 2 times a day by mouth with colostrum-milk; 0.2 g / kg body weight of the animal enterosorbent "Polysorb-VP" inside in the form of an aqueous suspension 1 hour before giving colostrum or 2 hours after feeding; reopoliglyukin with glucose 5 ml per 1 kg of live weight 1 time per day intravenously; 50 ml of isotonic sodium chloride solution per head intravenously 1 time per day. As a result, the animal suffered from mild dyspepsia.

Claims (2)

1. A method for predicting the development of a toxic form of dyspepsia in newborn calves with initial signs of dyspepsia, including determining platelet aggregation activity using ADP, collagen, ristomycin and adrenaline solutions as inducers by mixing on a glass slide 0.1 ml of platelet-rich plasma standardized for platelet count and 0.1 ml of one of the aggregating agents, and determining the time of manifestation of platelet aggregates in the form of whitish grains visible to the naked eye ohm, while accelerating platelet aggregation over 7% of the control, the development of a toxic form of dyspepsia is predicted. 2. The method according to claim 1, characterized in that the average result of determining induced platelet aggregation in ten clinically healthy newborn calves is used as a control.