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Method for preventing functional dyspepsia and physiological imunodeficiency in calves

Method for preventing functional dyspepsia and physiological imunodeficiency in calves
IPC classes for russian patent Method for preventing functional dyspepsia and physiological imunodeficiency in calves (RU 2269351):
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FIELD: veterinary obstetrics.

SUBSTANCE: the method deals with subcutaneous injection of bioglobin (placenta denaturated suspended - PDS) at the quantity of 20 mg/animal/d not earlier that 10 d before calving. The innovation enables to increase quality in preventing the onset of functional dyspepsia in newborn calves and level of total body resistance.

EFFECT: higher efficiency.

20 dwg, 1 ex, 2 tbl

 

The invention relates to the field of animal husbandry, in particular animal, and can be applied in the prevention of acute gastrointestinal illness in newborn animals.

A known method of preventing the occurrence of dyspepsia by the use of corticosteroids during the first days of life calves, which prevents the occurrence of diseases in 20,57% of cases (Alexandrov I.D. Corticosteroids for the prevention and treatment of calves with dyspepsia //W. Veerinary. - 1985. No. 1. - p.61-62).

The use of this method without preliminary studies of the hormonal status of animals can lead to negative consequences (Mashkovsky PPM Drugs /Hormones of the adrenal cortex. - 1993. - 1 h - 666 C.). In addition, these drugs do not possess antibacterial activity against pathogenic microorganisms, often translating dyspepsia from a simple form in toxic. The increasing levels of endogenous glucocorticoids after injection of the above drugs, only for a short time delay time to the development of clinical symptoms of dyspepsia and does not affect the concentration of catabolic hormones of the thyroid gland.

There is also known a method of increasing the resistance of newborn calves (SU 1801003 5, a 61 K 31/00, 1993), in which animals injected nonspecific immunoglobulin in complex with 2-t is metilaminoantipirina. This method does not provide the relationship between the levels of glucocorticoids and tireoide in the blood of newborns.

The objective of the invention is improving the quality of prevention of occurrence of functional dyspepsia in newborn calves.

This is achieved by the fact that as a preventive measure applied immunomodulator (EN 2036651 5, a 61 K 31/00, 1995) bioglobin (placenta denatured suspended - PDS). Bioglobin - tool, normalizes metabolic processes in the organism, which has immunomodulatory and adaptogenic effects due to the presence in its composition of polypeptides, amino acids and nucleic acids.

Due to the fact that newborn calves during the first days of postnatal period is a deficit of endogenous immunoglobulins and corticosteroids, which increases with dyspepsia, the introduction of myoglobin provides increased endogenous production by the adrenal cortex cow-woman and possibly fruit - cortisol, while reducing the level of thyroxine (Grishina ETC. Dynamics of thyroid hormones in neonatal diarrhoea of calves //R.zh. Veterinary - 1983. - No. 12 - s). Cortisol, in turn, increases the production of immunoglobulins and obviously immunoglobulin G, which passes through the placental barrier, also raises the level of natural resistance.

This comprehensive action is in General helps to restore the immune status of the organism, the function of the gastrointestinal tract and prevents the development of pathogenic microflora.

Example. Bioglobin was administered to cows at a dose of 20 ml/bird/day subcutaneously for 10 days before delivery.

Parturition in cows passed without complications in physiological terms.

They selected four groups of newborn calves (n=4) with functional (simple) form of dyspepsia: 1st group - calves without dyspepsia, from cows treated with myoglobin; 2-I - calves with dyspepsia from cows treated with myoglobin; 3-I - calves without dyspepsia, from intact cows (control 1); 4th - calves with dyspepsia from intact cows (control 2). Differential diagnosis excluded infectious diseases.

1) Blood for determination of cortisol and thyroxine immunoassay method and indicators of natural resistance: albumin; globulin alpha, beta, gamma and immunoglobulin took in calves from the jugular vein on the 2nd and 10th day after birth. Histological studies of immunocompetent organs of calves was performed on the 7th day (posthumously) after birth.

In the result noted (table 1)that on the 2nd day after birth the concentration of cortisol was highest in the groups of calves without dyspepsia: the 1st group - 78,3; in the 2nd - 50,75 nmol/L.

In calves with dyspepsia cortisol was significantly lower: in the 2-nd group - 27,15; 4th - 15,65 nmol/l decrease in the number cortisolemia 65,3% in the 2nd group compared to the 1st and 69.2% in the 4th, in relation to the 3rd group, testifies to activate steroidogeneza after treatment with myoglobin.

Increased levels of cortisol in calves without dyspepsia in the 1st and 3rd groups were observed at the 10-th day.

The level of thyroxine in groups of calves on the 2nd day after birth had different parameters.

The presence of high concentrations of free thyroxine in the blood of calves with dyspepsia 4-th group indicates the presence of hyperthyroidism. In calves the 3rd group (without dyspepsia) the amount of thyroxine is 36.5% less. And significantly fewer hormone observed in calves of the 1st and 2nd groups, where cows were injected bioglobin. The concentration of thyroxine in the 1st group was 2.3 times less compared to the indicators of the 3rd group, and 2nd, respectively, in at 16.1 times compared to the data of the 4th group.

The decrease of the hormone in the 1st group after treatment with myoglobin (7.9%) corresponds to that of the 3-th (25.5%; p<0.01), which indicates binormality influence of myoglobin during the time course of dyspepsia.

A significant increase of thyroxine in calves 2-th group, as well as the reduction of cortisol by the 10th day, may reflect the presence of deeper structural and metabolic disturbances in the body of the newborn or cow-mothers.

Table 1
Hormone levels in the blood of the new is born calves
№ p/p Group n=4 Cortisol, nmol/l Thyroxine, nmol/l
2nd day 10th day P 2nd day 10th day P
1. Calves without dyspepsia from cows treated with myoglobin 78,30±6,05 86,25±3,19 >0,2 62,00±12,30 57,13±6.42 per >0,5
2. Calves with dyspepsia from cows treated with myoglobin 27,15±4,00 25,55±2,48 >0,5 of 14.25±17,90 157,63±18,97 >0,5
3. Calves without dyspepsia from intact cows (control 1) 50,75±4,80 55,12±2,38 >0,5 146,25±6,25 109,00±6,01 <0,01
4. Calves with dyspepsia from intact cows (control 2) 15,65±0,88 16,42±0,75 >0,5 230,00±23,80 205,25±12,32 >0,2

2) Simultaneously conducted biochemical studies of humoral indicators of natural resistance in these calves showed (table. 2, the syndrome of diarrhea of newborn calves accompanied by a pronounced increase in serum albumin and alpha-globulins and decreased content of beta-globulin and gamma-globulin.

Thus, an excess amount of albumin in calves with dyspepsia compared with calves without it the 2nd day was: 2nd group - 27.2% greater than the value in the 1st group 4th group - 20.1% more than in the 3rd group. On the 10th day, this dependence is preserved and excess, respectively, was equal to: 2nd group - 19.8%4th group - by 21.0%.

Excess levels of alpha-globulin in these terms in calves with dyspepsia (2nd and 4th group) over the figures obtained in the 1st group, respectively, was: 6,7 and 10.0%. Even in calves without dyspepsia from intact cows had an excess amount of alpha-globulin by 3.6% compared with the data on the animals of the 1st group. On the 10th day concentration in the blood of alpha-globulins in the 1st group has decreased by 29.7% and was lower than the 2nd, 3rd and 4th groups: 25,6; 44,8 and 24.7%.

The content of beta-globulin on the 2nd day from birth was lowest in calves 4th (calves with dyspepsia from intact cows) group. The decline compared with the data of the 1st, 2nd and 3rd groups were: 10,4; is 24.4 27.7%). A slight increase (14.1%) on 10-th day little changed the situation and the level of beta-globulins remained reduced by 21.4% compared to what the value is in the 3rd group (calves without dyspepsia from intact cows). The same pattern is noted and the number of gamma-globulin. Most of them on the 2nd day of the 1st group is significantly higher than the values in the 2nd, 3rd and 4th groups: 55,0; 43,5 and 56.4%, respectively. On the 10th day, the ratio is even more increased with respect to the 4-th group, and similarly was: 40,2; 50,2, and 70.0%.

Table 2
Biochemical parameters of blood of newborn calves
Group (n-4) Immunoglobulins% Albumin Globulins
α 3 1
2nd day 10th day 2nd day 10th day 2nd day 10th day 2nd day 10th day 2nd day 10th day
1. Calves from cows treated with myoglobin without dyspepsia 13,65±0,92 17,42±1,51 35,14±1,45 34,08±1,00 of 18.45±0,31 16,40±1,10 16,50±1,01 16,87±0,95 29,90±of 2.51 32,95±1,50
2. Calves from cows treated bioglobin the m with dyspepsia 5,96±0,24 by 5.87±0,40 41,46±0,63 39,02±, and 0.40
p<0,02
16,69±0,49 18,50±0,94 19,57±0,57 19,00±0,29 19,28+1,20 23,49±1,36
3. Calves from cows without treatment, without dyspepsia 7,47±0,41 8,39±0,38 39,49±0,76 37,84±0,35 19,23±0,51 18,74±0,72 20,45±0,30 21,49±0,52 20,80±1,48 21,93±0,50
4. Calves from cows without treatment dyspepsia 1,37±0,26 1,32±0,19 45,83±1,05 44,50±0,77 20,30±2,40 19,20±1,20 14,80±0,85 16,90±0,78 19,11±1,04 19,38±0,96

The ratio a/G also had values that match the physiological condition of the animals. So in the 2nd and 4th groups, the ratio A/G was the highest, respectively: on the 2nd day of 0.7 and 1.03; on the 10th day to 0.63 and 0.80. In the 1st and 3rd groups, where calves were without dyspepsia, ratio of, respectively: on the 2nd day of 0.54 and 0.65; 10-th day - 0.51 and 0,60.

The best indicator on the 2nd day of the 1st group, 20.3% less than in the 3rd group and 29.6% less from what azaela calves 2-th group and at 90.7% less than the value of A/G in the 4th group. On the 10th day total protein ratio of coefficient groups had the same orientation and decreased slightly, mainly due to a small increase in the number of globulins.

Thus, blood biochemical parameters indicate pronounced binormality the influence of myoglobin aimed at reducing physiological immunodeficiency newborn calves.

3) Histological studies were performed on three groups of newborn calves (n=3). 1st group - calves without dyspepsia from cows-mothers treated myoglobin at a dose of 20 ml/bird/day, for 10 days prior to delivery. Group 2 calves without dyspepsia from intact cows-mothers; group 3 calves with dyspepsia from intact cows-mothers. Pathological material (mesenteric lymph nodes, spleen, thymus) for histological studies according to standard techniques, was selected posthumously on the 10th day after birth.

The result was installed following picture histomorphologically changes in immunocompetent organs.

Spleen

1st group. Quantitatively lymphoid tissue of the spleen calves developed about the same as calves of the 2nd group. However, the cellular composition of the white pulp has significant features. Most splenic cells (Figure 1 (2)) are reactive centers (Figure 1, (4)), among the cells which Phi is .1 (1)) meet macrophage and mitotically active elements, apparently proliferating b-lymphoblasts. Marginal zone splenic cells, on the border between the white and red pulp (Figure 1 (3)), is moderately populated by small lymphocytes and contains isolated macrophages. Around the pulp vessels are observed uniform accumulation of small lymphocytes, forming a periarterial lymphatic sheaths.

The above gives grounds to speak about the preferential activation In dependent areas of the white pulp of the spleen in calves 1-th group.

In the red pulp, compared with calves 2nd group more visible pulp fibers, often with blast cells and macrophages. The sinuses of the red pulp moderately filled with blood.

On the stimulating effect of myoglobin on the elements of the immune protection testify extensive diffuse accumulations of lymphoid tissue in the mucosa of the duodenum 12 and abomasum of calves (Figure 2(5); 3(1)). In places these clusters are nodular in nature. Some nodules are formed reactive centers (Figure 2 (6)).

The 2nd group. Microscopic characteristics of the body indicates a typical structure. Lymphoid tissue in the white pulp occupies about 20% of the parenchyma. The red pulp is moderately filled with blood. White pulp presents emerging splenic cells and perivascular lymphoid couplings (Figure 4 (7)). And the calf and the clutch are the C tightly Packed lymphocytes, without their zonal location. Between lymphocytes were not detected macrophage elements and blast shape of the cells (Figure 5).

Reactive centers in moderately developed follicles were not found.

These features white pulp show somewhat greater development of T-dependent zones and minor functional activity-dependent areas of the body.

Between weakly crownpointe the sinuses of the spleen revealed the pulp fibers of the body, poor as blast elements, and macrophages.

3rd group. In calves in this group for histological structure in the spleen marked anaplasia lymphoid tissue friability and the absence of zoning in the splenic corpuscles and blood sinusoids red pulp (6 (8)). In reticular stroma often found macrophage elements.

In the spleen only occasionally formed lymphoid cells and perivascular clutch. In the center of some Taurus, you may notice a slight loosening, morphologically resembling reactive centers (6). Basically, poor lymphoid tissue is diffuse on the background of moderately cravenplan of sinusoidal red pulp.

For spleen calves in this group on the background of immunodeficiency States the most notable weakness In dependent areas of the white pulp.

<> The thymus

1st group. For the thymus gland of calves in this group are characterised by fairly expressed involutive processes. However, in the lobules well contribut cortex and the medulla. In the cortical substance of the densely arranged lymphocytes mask cells of the stroma. Often you can find figures of mitosis blast lymphoid elements. Rare perforated education and destructive modified lymphocytes. However, relatively frequently found active macrophages.

In the medulla is relatively much timoszyk Taurus different generation. Bullock, as a rule, thick, juicy, with a small number of destructive changes.

The whole body is characterized by immunodeficiency condition at level I, II degree, according to the existing classification.

The 2nd group. The thymus gland of calves in this group is typical for the species and age of the animal structure (Fig.7). The connective tissue capsule thin layer that separates the body into slices. In strips of well contribut blood vessels are rare macrophages and eosinophils, as well as cells plasmacytoma series.

In each slice well different cortical (Fig.7(9)) and the medulla (Fig.7 (10)). Cortical substance contains a large number of relatively densely Packed small lymphocytes. Between them noticeable e the cops epithelial reticular skeleton, and, particularly in podkapsuliarnaya area, larger lymphoblasts. The cortical substance is almost 2/3 of the area of each slice slices. Closer to the border with the substance of the brain meet destructive modified lymphocytes events karyorhexis.

The medulla is lighter due to less content of lymphocytes in the loops of the cytoplasm of epithelialisation. Last sometimes as if isolated, their cytoplasm becomes more oxytelinae, and the core is lighter, puzarchaty. This is already the unicellular stage of the future layered epithelial timusewo Taurus. Such a calf is found closer to the Central part of the brain lobes of the thymus. Near timoszyk Taurus some macrophages and acidogenicity. In some cells there are destructive changes.

3rd group. For the thymus gland of calves in this group were characterized by extreme involutive processes and immunodeficiency. The body had no signs of dividing the slices on the cortex and the medulla. Observed loosening of the parenchyma and stroma of the slices. Over the entire area of the slice met timoshii bullock unicellular stage (Fig (11)). Rarely found multicellular epithelial bullock without clear outlines. In them, as a rule, was marked by a pronounced destructive changes and deathly the pockets. However, macrophage reaction in the body was moderate (Fig, 9 (12), 10(12), 11(12)).

Lymph nodes

Group 1. In the cortical substance of the body over the entire area of the pyramidal region detected follicles of different sizes (Fig(13)). The most clear follicles are smaller, rounded in shape, with well-defined centers of reproduction. Closer to pottapalayam the sine meet the larger aggregations of small lymphocytes, but with less pronounced breeding centers (Fig(14)).

There is a relatively well-developed paracortical area of the body, saturated with small lymphocytes (Fig(15)). From this zone in the depth of the body are well using brain fibers (Fig(16)) with a large number of lymphocytes, among which there are immunoblastic (Fig(17); 15(17)). Wide cerebral sinuses filled free cellular elements (Fig(18)), especially macrophages, and clearly contour reticular cells with hyperchromic nuclei (Fig(19)).

Group 2. In calves in this group mesenteric lymph node has a typical structure (Fig). In the cortical substance can be distinguished pyramidal region with follicles of different sizes (Fig(13)) and moderate breeding centers. On the periphery of the follicle cells are more densely, forming concentric zone. In the centres is esmerine you can distinguish immunoblastic and dendritic reticular cells. Paracortical area relatively rich diffuse located in small lymphocytes. The medulla site presents well-developed strands, rich small lymphocytes. Occasionally blast the elements.

Cerebral sinuses contain small, encouragethe reticular cells, a small number of lymphocytes and a single in sight macrophages (Fig).

Group 3. In the lymph nodes of the pyramidal region of the cortical substance is not expressed. Lymphoid tissue contains a very small number of lymphocytes (Fig(14)) in the loops dendritic reticular cells. There are no formed follicles. Minor compacted areas of lymphoid tissues do not contain centers of reproduction (Fig). Given that the latter are the location of lymphocytes into plasma cells, it is possible to speak about a significant weakening of the humoral immunity of the organism.

Paracortical area of the body is also poor in lymphocytes. It is the main thymus-dependent area of the lymph nodes. Its weak development shows a decline in cellular immune protection.

The inner zone of the cortical substance of the lymph node becomes weakly using the fibres of the brain substance with a low content of lymphocytes and immunoblasts, which is also a sign of immunosuppressive condition. In the relatively near the fir sinuses of the brain substance found little free cellular elements - lymphocytes and macrophages (Fig).

Thus, processing myoglobin cows-mothers for 10 days before the birth helps to activate the immune status of the newborn and the reduction in the physiological immunodeficita calves due to activation steroidogeneza, immunoglobulins, as well as positive morpho-functional changes in immune organs of animals.

The use of myoglobin at a dose of 20 ml/bird/day, for 10 days the cow-mothers helps prevent the occurrence of physiological dyspepsia in 90% of cases. Efficiency is 1 ruble costs 63 roubles of profit.

Sources of information

1. Alexandrov I.D. Corticosteroids for the prevention and treatment of calves with dyspepsia //veterinary medicine. - 1985. No. 1. - p.61-62.

2. Grishina SO, the Dynamics of thyroid hormones in neonatal diarrhea in calves / & W. Veterinary science. - 1983. No. 12. - S. 25.

3. Mashkovsky PPM Drugs /Hormones of the adrenal cortex. - 1993. - 1 h - 666 S.

4. EN 2036651 5, A 61 K 31/00, 1995.

5. SU 1801003 5, A 61 K 31/00, 1993.

The way to prevent functional dyspepsia and physiological immunodeficiency syndrome in calves, namely, that before the birth of cows injected immunomodulator bioglobin (placenta denatured suspended - MPD) subcutaneously at a dose of 20 mg/bird/day not more than 10 days.

 

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