ÓÄÊ 619:636.2:591.16:591.11

CLINICO-HEMATOLOGIC AND BIOCHEMICAL STATUS OF COWS AT GESTOSIS

A.G. Nezhdanov, M.I. Retskii, Yu.N. Alekhin, V.A. Safonov, V.I. Shushlebin, N.E. Papin, T.P. Brekhov, E.V. Shishkina

The authors evaluated the clinical, hematologic, biochemical and hormonal status of highly productive dairy cows at gestos. It is established, that the degree of expressiveness of changes hormonal-hematologic homeostasis in animals and their clinical state depends on weight of current of pathological process. The double injection to cows of the selemag preparation, containing selenium, normalizes hemopoiesispoes, a metabolism, functional activity of fetoplacental complex, cardiovascular, secretory and reproductive systems.

Key words: cows, gestosis, clinical, hematological, biochemical, hormonal parameters, action of selemag.

 

Gestosis, or the sickness of pregnancy adaptation (pre-eclampsia), is an actual problem of highly-productive dairy cattle (1-4). The pathology develops owing to inability of mother adaptive systems to meet the needs of developing fetus, which is manifested as changes in immune, endocrine and antioxidant status of mother organism and the varying degrees of perfusion-diffusion placental insufficiency (5-7). The pathology process involves fetoplacental complex, cardiovascular and pulmonary system, liver, kidney and central nervous system (2, 8). At the same time, fetal development disturbs, fetal viability reduces, and severe complications at calving and in postpartum period take place as well (4, 9).

Despite the significant interest to the problem of gestosis in animals, the genesis of this disease still remains poorly understood, which makes difficult its early diagnosis, prevention and therapy.

The purpose of this study was to identify the peculiarities of hematological, biochemical and hormonal status in highly-productive dairy cows at the gestosis of different severity.

Technique. The experiment was performed in the breeding farm Plemzavod "Druzhbah" (Pavlovsky district, Voronezh province) in 2009. The objects were 120 pregnant dry cows the Red-and-White breed aged 4-7 years and weighing 550-600 kg, whose average annual milk productivity was 6,7 thousand kg. Feeding rations of the animals corresponded to the standards of All-Russia Research and Development Institute of Animal Husbandry (10).

Upon the results of clinical examination (evaluation of general condition, monitoring of blood pressure, identification of body swelling and protein in urine), the animals were divided into 3 groups: clinically healthy cows, cows with complete gestosis syndrome (arterial hypertension, proteinuria and body swellings) and cows with individual symptoms of gestosis.

The venous blood samples were taken from 8-10 individuals of each group to determine the contents of erythrocytes, leukocytes, platelets, hemoglobin, fibrinogen, molecules of average mass (MAM), hematocrite, protein, urea, creatinine, glucose, lactic acid , total lipids, cholesterol, triglycerides, malondialdehyde (MDA), carotin, vitamins A, E and C, total calcium, inorganic phosphorus, iron, magnesium, copper, zinc, manganese, ALT (alanine aminotransferase), AST (asparte ataminotransferase ), g-GT (g-glutamiltransferase), ALP (alkaline phosphatase) (11), as well as the contents of progesterone, estradiol-17?,  testosterone, cortisol (by enzyme immunoassay - ELISA test-systems by “DVC”, USA).

50-55 and 25-30 days before the expected calving, cows were injected with selenium-containing preparation Selemag. 5-10 days before calving, the repeated blood tests and evaluation of animals’ clinical state were performed. The character of calving process and a postpartum period were considered as well.

The obtained data were processed statistically using Statistica 5.0 software.

Results. The development of pregnancy pathology in cows was accompanied by significant changes in their clinical and hematological status, while the degree of their manifestation depended on severity of pathological process.

Morphological, biochemical and hormonal parameters of blood in late-pregnant dairy cows the Red-and-White breed depending on severity of gestosis pathology (Õ±õ, Pavlovsky district, Voronezh province, 2009)

Blood parameter

Group of cows

Clinically healthy

Slight gestosis

Severe gestosis

Hematological status

Erythrocytes, 1012/l
Leucocytes, 109/l
Neutrophils, %
Eosinophils, %
Monocytes, %
Lymphocytes, %
Platelets, 109/l
Hematocrite, %
Sorption capacity of erythrocytes, %

5,7±0,13
8,4±0,47
32,0±4,14
9,3±2,02
2,9±0,34
57,1±2,91
404,1±25,70
37,5±0,56
35,0±1,03

5,9±0,27
8,7±0,34
32,0±1,50
8,6±1,69
2,5±0,45
57,7±1,41
324,8±27,02
39,1±0,56
43,7±1,00

5,8±0,11
9,1±0,32
29,7±2,73
14,4±3,10
2,6±0,71
54,2±2,48
286,5±8,56
38,1±0,62
54,5±2,25

Biochemical status

Total protein, g/l
Urea, mmol/l
Creatinine, umol/l
Total lipids, g/l
Cholesterol, mmol/l
Triglycerides, mmol,l
Malondialdehyde, umol/l
Glucose, mmol/l
Lactic acid, mmol/l
Alanine aminotransferase, U/l
Aspartate aminotransferase, U/l
g-Glutamyltransferase, U/l
Alkaline phosphatase, U/l
Carotin, umol/l 
Vitamin A, umol/l
Vitamin E, umol/l
Vitamin C, mmol/l
Total calcium, mmol/l
Inorganic phosphorus, mmol/l
Iron, mg%
Magnesium, mg%
Copper, ug%
Zinc, ug%
Manganese, ug%

79,8±1,56
3,4±0,23
101,4±7,50
4,1±0,35
3,5±0,27
0,5±0,02
1,1±0,09
2,2±0,14
1,8±0,07
27,5±1,72
58,5±1,46
14,9±1,31
91,0±3,93
0,2±0,02
1,2±0,10
11,2±0,89
14,5±5,73
2,3±0,02
2,1±0,05
21,8±0,17
2,4±0,07
90,2±4,39
272,7±9,82
10,5±0,45

82,2±0,82
3,4±0,26
97,5±5,08
3,1±0,18
3,7±0,23
0,6±0,02
1,2±0,08
2,6±0,17
2,4±0,10
30,9±3,17
66,3±5,11
15,5±0,25
121,2±16,54
0,2±0,02
1,2±0,09
9,9±1,20
18,1±4,02
2,5±0,04
2,1±0,06
21,4±0,34
2,4±0,08
91,8±2,37
289,8±19,98
11,8±0,66

80,6±2,08
3,1±0,19
99,6±3,35
3,5±0,26
3,6±0,30
1,1±0,04
1,3±0,15
2,4±0,11
4,5±0,13
29,2±3,27
58,2±2,38
14,6±1,13
85,1±13,39
0,2±0,01
1,3±0,15
7,7±0,93
12,0±1,69
2,5±0,03
1,9±0,07
20,9±0,22
2,6±0,28
78,4±3,22
258,2±16,08
1,0±0,38

Hormonal status

Testosterone, ng/ml
Progesterone, ng/ml
Estradiol-17?, pg/ml
Cortisol, ng/ml

1,3±0,22
24,7±4,62
273,4±38,40
32,7±5,79

0,7±0,09
10,5±2,09
215,2±17,90
24,4±3,01

1,2±0,33
11,2±4,31
270,1±25,40
21,4±3,47

The first clinical symptoms of developing gestosis were: increased cardiac activity, raise of blood pressure, abnormally rapid breathing - i.e. the reaction aimed at maintaining the disturbed uteroplacental blood flow within physiological limits. Thus, at the initial stage of preeclampsia (slight disease), pulse frequency in cows increased from 64,8 ± 4,08 up to 81,0 ± 2,63 bpm, or 25,0% (P <0,001) and respiration rate - from 18,5 ± 0,61 up to 20,8 ± 0,94, or 12,4%, the level of systolic blood pressure (BPsys) - from 105,3 ± 1,63 to 118,7 ± 3 01 mmHg, or by 12,7% (P <0,001) and diastolic (BPdias) - from 63,3 ± 1,73 up to 69,1 ± 3,01 mmHg, or by 9,2%.

Protein content in the urine of clinically healthy animals was 0,6 ± 0,07 g/l, while in cows with gestosis symptoms - 1,1 ± 0,44 g/l (increased by 83,3%), and urine pH increased from 7,4 ± 0,10 to 7,6 ± 0,09.

When the development of complex gestosis syndrome in cows (severe disease), their heartbeat rate increased to 86,8 ± 3,84 beats/min (34,0% higher than in clinically healthy individuals), BPsys - up to 136,1 ± 2,85 mmHg (29,2% higher), BPdias - up to 83,5 ± 3,22 mmHg (31,9% higher). The respiration rate remained unchanged. Protein content in the urine increased to 3,0 ± 0,49 g/l, or 5-fold (P <0,01), urine pH - up to 8,0, or 8,1%.

The development of preeclampsia in cows was accompanied by morphological changes in the blood (see table). The total number of leukocytes increased by 9,0%, eosinophils - by 54,1%, lymphocytes - decreased by 5,4%, neutrophils - by 7,7%, platelets - by 41,0% (P <0,01), hematocrit increased by 4,3%, sorption ability of erythrocytes (SAE) - by 55,7% (P <0,001).

The detected changes in blood composition along with the increased content of MAM from 0,2 ± 0,01 to 0,3 ± 0,03, or 60% (P <0.01) reflected the enchasing syndrome of endogenous intoxication, whose manifestation depended on severity of pathological process.

The development of thrombocytopenia combined with increasing content of fibrinogen from 5,1 ± 0,65 to 6,1 ± 0,27 g/L, or 20%, indicated the raise of blood agglutination.

The biochemical blood test has revealed (See table), that the development of gestosis in cows was associated with decrease in concentrations of total lipids by 32,4, vitamin E - by 12,7% (P <0,05), the raise in content of triglycerides by 13,5, activity of ALT - by 12,4, AST - by 13,3 and ALP - by 33,2%. These data suggest that the liver, kidneys and placenta started being involved into the expanding pathological process.

Compensatory reactions of an organism to progressing pathological process manifested themselves as normalization of activity of transamination enzymes. At the same time, the continuation of functional disorders in the liver and kidneys was revealed by the decrease of urea in the blood by 12,1%, the ratio between concentrations of urea and creatinine - by 14,6%, and the increase in triglycerides content in 2,1 times (P <0,001).

Already in early stage of gestosis process, lactic acid content in the blood increased by 33,3% (P <0,001) and vitamin E – decreased by 11,3%, which was connected with activation of free-radical oxidation and antioxidant defense system.

The severe stage was characterized by the raised content in the blood of intermediate products of lipid peroxidation – MDA by 20,0%, lactic acid - in 2,5 times, the decline in content of vitamin E by 45,8 (P <0,05), vitamin C - 12,1, copper - by 15,0 and zinc - by 5,6%.

The obtained data indicate that pregnancy pathology in cows clinically manifested as the complex of gestosis symptoms develops against a background of increased lipid peroxidation (decrease of contents in blood of total lipids, vitamin E, increase - MDA), the accumulation of toxic metabolites (increase in MAM content and eosinophilia), enhancing the  processes of blood agglutination (thrombocytopenia, reduced activity of  fibrinolysis), liver function abnormalities (decrease in blood of urea and vitamin E contents, increase in concentrations of cholesterol and triglycerides, the raised activity of g-glutamyltransferase and alkaline phosphatase), kidney (proteinuria, decrease in calcium content in the blood, the ratio urea / creatinine). It can be assumed that a slight gestosis is accompanied with inflammatory processes in an organism, while dystrophic processes occur at a severe preeclampsia.

The development of anemia (reduction in concentrations of copper and zinc in the blood, increasing the permeability of erythrocyte membranes) is another gestosis symptom.

The evaluation of endocrine status of the cows has revealed the development of gestosis occurs against a fetus-placental insufficiency, which was indicated by the low content of all studied hormones. The concentration of testosterone in the blood of these animals was 1,7 times lower than in clinically healthy ones, progesterone – in 2,4 (P<0,05), estradiol-17b and cortisol - in 1,3 times (see Table).

In other words, the activation of compensatory mechanisms in the fetus-placental complex leads to the increased synthesis of testosterone and estradiol and the rise of their contents in blood up to the levels of clinically healthy animals, while progesterone and cortisol levels remained low. These data are consistent with results of earlier studies (12).

The two-time injection of the selenium preparation Selemag was accompanied by normalization of hematopoiesis, biochemical and hormonal status. Thus, the number of leukocytes in blood reduced from 9,1±0,32I109/L to  7,4±0,10I109/l or 18,9 %, while the number of neutrophils increased by 12,8 %, lymphocytes - by 7,7 %, platelets - by 12,5 % and the number of eosinophils decreased in 2,1 times (P <0,05), hematocrit – by 3,5 %, sorption capacity of red blood cells - by 4,8 %.

MDA content decreased by 1,1 ± 0,09 mmol/l (or by 17,8 %), MSM - up to 0,3 ± 0,03 conv. units. (by 23,1 %), glucose - increased to 3,0 ± 0,15 mmol/l (by 28 %), vitamin E - up to 12,3 ± 3,32 umol/l (in 1,50 times), vitamin C - up to 27,8 ± 6,35 mmol/l (in 2,32 times), zinc - by 32,8 %, manganese – by 21,2 %, copper - by 11,4 %, selenium - from16,7 ± 1,41 up to 29,8 ± 2,58 ug% (or 78,4 %), calcium - decreased by 10,6 %.
Consequently, the selenium preparation has shown its normalizing effect on blood forming in cows, as well as on the reduce of lipid peroxidation activity, increase of antioxidant status, activation of miroelements metabolism, energy processes and the relief of endogenous toxicosis.

These facts were clinically manifested as the normalization of cardiovascular functional activity (BPsys decreased by 8,2%, BPdias - by10,9%) and secretory systems (decrease of the protein content in urine by 37,4%).
The preparation also activated endocrine function of placenta: the concentration of estradiol-17β in blood increased by 404,5 ± 108,20 pg/ml, or in 1,45 times, progesterone - up to 20,0 ± 10,77 ng/ml , or in 1,78 times, and cortisol - up to 29,7 ± 2,52 ng/ml, or in 1,39 times.

Normalization of hormonal-metabolic status of cows with a symptom of preeclampsia provided by double injection of Selemag allowed to reduce the frequency of calving complications and postpartum pathology in 1,22 times.

Thus, the development of gestosis in cows includes a complex of changes in hematological, biochemical and hormonal status, which differ by phases of pathological process, depend on its duration and peculiarities of compensatory-adaptive reactions. Gestosis pathology is accompanied by activation of peroxidation processes, the raise of endogenous intoxication against a background of placental insufficiency and nephrotoxicity as a permanent component. Therefore, gestosis prevention and treatment in cows should be aimed at normalization of lipid peroxidation, rheological and coagulation blood properties, the functions of fetoplacental complex as well as kidney and liver.

REFERENCES

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8. Strizhakov A.N., Davydov A.I. and Belotserkovtseva L.D., Izbrannye lektsii po akusherstvu i ginekologii (Selected Lectures on Midwifery and Gynecology), Rostov-na-Donu, 2000.
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 All-Russia Research and Development Veterinary Institute of Pathology, Pharmacology and Therapy,
Russian Academy of Agricultural Sciences, Voronezh 394087, Russia,
e-mail: vnivipat@mail.ru

 

Received January 11, 2010

 

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