Method to forecast development of toxic form of dyspepsia in newborn calves with initial symptoms of dyspepsia

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

 

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

It is now established that in certain pathological conditions of animals to observe changes in the aggregation activity of platelets.

There are various methods of determining the functional capacity of platelets, based on the assessment of spontaneous or induced (using various agonists) platelet aggregation (Barkagan SS, Momot A.P. Diagnostics and controlled therapy of disorders of hemostasis. - M.: "Novamed-JSC, 2001. - 296 S.).

A known method of determining aggregation activity of platelets on glass" (Shitikova AS. Visual micromethod determine the aggregation activity of platelets with various aggregating agents / Ashitkov // Laboratory work. - 1984. No. 4. - S-220).

However, all these methods are only used to assess the functional activity of platelets for various diseases, and to predict the development of toxic forms of dyspepsia are not applied.

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

Early prediction of the development of toxic dyspepsia newborn calves is of great practical importance because it allows a more thorough approach to t is rapee and timely preventive treatment of sick animals, that will help reduce the mortality of cases of dyspepsia newborn calves and raising the economic efficiency of livestock.

The essence of the method consists in the following. Newborn calves with an initial signs of dyspepsia take samples of blood and determine aggregation activity of platelets using as inductors solutions ADP, collagen, ristomycin and adrenaline by mixing on a slide 0.1 ml of standardized platelet count platelet-rich plasma and 0.1 ml of one of the aggregating agents, and identify the time of manifestation of platelet aggregates in the form of whitish grains visible to the naked eye. The control used is the average result of the determination induced platelet aggregation in 10 clinically healthy calves. In the detection of newborn calves with an initial signs of dyspepsia acceleration of platelet aggregation over 7% of control it is possible to predict the development of toxic dyspepsia.

The method is as follows: newborn calves with an initial signs of dyspepsia make sampling blood in the morning before feeding in plastic dishes. To prevent spontaneous aggregation apply the stabilization of the obtained blood of 3.8% solution of sodium citrate is a ratio of 1:4. Define induced aggregation activity of platelets using as inductors solutions ADP (breeding stock solution 1:10), collagen (concentration 20 mg/ml), ristomycin (concentration 15 mg/ml) and epinephrine (dilution of stock solution 1:50). From stabilized blood receive platelet-rich plasma and platelet-poor plasma. Used platelet-rich plasma obtained by centrifugation of the obtained stable blood at 1000 rpm for 6 min; and plasma poor in platelets, after centrifugation the remaining in vitro platelet-rich plasma at 3000 rpm for 20 minutes platelet-Rich plasma standardise the number of platelets by dilution, if necessary, autologous sample of platelet-poor plasma (up to 380-400000 platelets/µl). The volume mix is rich and platelet-poor plasmas is determined by the formula (1):

where Vppp- the volume of platelet-poor plasma,

Vprp- the volume of platelet rich plasma,

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

Further, different pipettes for non-fat dry glass slide put 0.1 ml of the investigated platelet-rich plasma and 0.1 ml of one of the aggregating agents. In the Les of mixing with a glass rod drops on a glass slide immediately include a timer and set the time of manifestation of the so-called phenomenon of the snow storm when aggregates of platelets reach such sizes that can be visible to the naked eye as whitish grains (required the use of light microscope type OI-19). The phenomenon of snow storms in healthy individuals occurs at the same time, while strengthening the aggregation function it appears earlier standards, with the weakening later. Response repeat 2 times with the same aggregation agent. The results take into account in seconds. The result is considered average from the two definitions of the values.

The control used is the average result of the determination by the above method induced platelet aggregation in 10 clinically healthy calves of the same age group.

In the detection of newborn calves with an initial signs of dyspepsia acceleration of platelet aggregation over 7% of control it is possible to predict the development of toxic forms of dyspepsia.

Example 1. According to the principle of analogues with regard to the origin, age and body weight at birth were formed 2 groups of newborn calves. Terms of housing, feeding and care of animals were the same. The first group consisted of 22 clinically healthy calves. The second group was composed of 17 calves with initial symptoms of dyspepsia were treated according to the sh farm is IU (tylosin-50 0.2 ml/kg intramuscularly 1 time per day until recovery; the broth sorrel isotonic sodium chloride solution inside the morning and evening at 0.5 l). During the experiment the second group was divided into two subgroups: patients with simple (n=10) and patients with a toxic form of dyspepsia (n=7) calves. The criterion for this division was the difference in the clinical picture of the disease. For calves with simple dyspepsia was characteristic not too rapid defecation emitting liquefied feces, the General condition of patients calves remained satisfactory. In calves, sick toxic form of dyspepsia, with full development of the disease pattern was observed profuse diarrhea, were observed depression, weakness, emaciation, therasense coat, dryness of nasal mirror.

In patients with dyspepsia calves (n=17) platelet aggregation was determined in the beginning of the disease (when the first signs of dyspepsia), in the midst of disease (at 2-3 days dyspepsia) and stage of recovery.

The results of determining induced platelet aggregation in clinically healthy and patients with dyspepsia calves are presented in table 1. As can be seen from table 1, patients with dyspepsia newborn calves at the beginning of dyspepsia in comparison with the average data of clinically healthy calves occurred acceleration induced aggregation on 5,51% when using collagen, 6,26% when s is the physical alteration of ADP, on 5,95% when using ristomycin and 5,80% when using adrenaline. When toxic dyspepsia early in the disease the results of the aggregation was ahead of the average of the healthy newborn calves on 7,72% when using collagen, 8,39% when using the ADF, of 7.82% in ristomycin and 8,09% when using adrenaline.

Table 1.
Induced platelet aggregation (AT) clinically healthy and patients with dyspepsia newborn calves
InductorsAverage AT a healthy newborn calves (n=22)Simple dyspepsia (n=10)Toxic dyspepsia
OnsetThe height of the diseaseStage recoveryThe onset of the disease (n=7)The height of the disease (n=7)Stage of recovery(n=5)
Collagen,29,92±1,16of 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,38,97±1,6136,53±0,39*32,94±0,07*37,06±0,07*35.70 barm±0,40*to 23.03±0,06*34,13±0,14*
Ristomycin,41,02±1,6138,58±0,37*36,17±0,06*39,49±0,08*37,81±0,39*24,11±0,10*36,33±0,18*
Adrenaline97,05±4,0191,42±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 with the average AT a healthy newborn calves

Accordingly, already at the early symptoms of dyspepsia, when it is impossible to diagnose the disease, platelet aggregation occurs before more than 7% of those newborn calves, which further perebolevayet toxic form of dyspepsia. That is they way the definition of induced platelet aggregation in the newborn calves on the proposed methodology can be used as a relatively simple and readily available in economy a method for predicting the development of toxic forms of dyspepsia in newborn calves with an initial signs of dyspepsia.

Example 2. The newborn calf No. 122 identified the initial symptoms of dyspepsia: not too rapid defecation emitting liquefied feces. To evaluate the risk of toxic forms of dyspepsia the animal selected blood sample, defined aggregation activity of platelets using as an inducer of collagen. This figure from a diseased calf equal 27,25 C. as a control used the average result of the definition of the aggregation activity of platelets in 10 clinically healthy calves (30,05). The acceleration of platelet aggregation from a diseased calf is 2,80 with or to 9.32%. Accordingly, this animal can predict the development of toxic forms of dyspepsia. The calf assigned adequate therapy: 0.1 ml of 0.1%solution of the drug "Biopag-D" 1 kg live weight of calves 2 times a day orally with colostrum-milk; 0.2 g/kg animal body weight of enterosorbent "Polisorb-EP" inside in the form of an aqueous suspension for 1 hour before giving colostrum or 2 hours after feeding; roopali who Liukin with glucose 5 ml per 1 kg body weight of 1 times a day intravenously; 50 ml of isotonic sodium chloride solution on the head intravenous 1 time per day. In the result, the animal had indigestion in the form of light.

1. A method for predicting the development of toxic forms of dyspepsia in newborn calves with an initial signs of dyspepsia, which includes the determination of the aggregation activity of platelets using as inductors solutions ADP, collagen, ristomycin and adrenaline by mixing on a slide 0.1 ml of standardized platelet count platelet-rich plasma and 0.1 ml of one of the aggregating agent, and determining the time of manifestation of platelet aggregates in the form of whitish grains visible to the naked eye, when the acceleration of platelet aggregation over 7% of control predict the development of toxic forms of dyspepsia.

2. The method according to claim 1, characterized in that the control used is the average result of the determination induced platelet aggregation ten clinically healthy calves.



 

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