RU2246309C1 - Method for prophylaxis of chlamydium bronchopneumonia in calves - Google Patents

Abstract

FIELD: veterinary science, infectious diseases.

SUBSTANCE: method involves applying furacyclin M as a tetracycline preparation and probiotic -ocarin additionally for 10 days that are given to calves from 12 old days age by the following schedule: furacyclin M in the dose 0.4-0.5 g/kg of body mass, 2 times per 24 h, and ocarin in the dose 8 x 10⁶ bacterial cells, 1 time per 24 h before night. Invention promotes to enhancing functional activity of blood neutrophiles, elevating content of T- and B-lymphocytes and activating their function that, in turn, promotes to enhancing the natural resistance of calves body. Invention can be used for prophylaxis against chlamydia bronchopneumonia in calves.

EFFECT: improved method for prophylaxis.

5 tbl, 2 ex

Description

The invention relates to veterinary medicine, relates to a method for the prevention of chlamydia, in particular chlamydial bronchopneumonia of calves, and can be used to increase the safety of young cattle.

Chlamydia is a group of contagious diseases of mammals, birds, and humans caused by antigenically related and morphologically similar microorganisms - chlamydia.

Chlamydia is characterized by pathogenesis and clinical manifestation as respiratory, enteric, urogenital, articular, neurological, ocular, as well as general diseases, affecting all organs and systems. They occur acutely, chronically and asymptomatically, and patients remain chlamydia carriers for a long time.

Chlamydial bronchopneumonia (a respiratory form of the disease) often occurs in young animals up to 6 months of age. According to some data, with respiratory infections, chlamydial bronchopneumonia is recorded in 70% of cases. The disease proceeds in the form of enzootia, mainly calves from 15-20 days of age and older fall ill due to infection of the respiratory tract against a background of low humoral (colostral) and cellular immunity. The disease is often complicated by a second (secondary) infection due to the activation and manifestation of the pathogenic effects of microorganisms - inhabitants of the upper respiratory tract or translocated from the intestine against the background of a weakening of the body's barrier functions. In this case, bronchopneumonia proceeds in severe form with a fatal outcome of up to 30-40% and above.

Known methods of therapeutic and prophylactic intervention for chlamydia, including the use of tetracycline drugs with chlamydiostatic action, in particular biomycin in chlamydial bronchopneumonia (1).

Fodder antibiotics of the tetracycline group (biovit, biotetracorm) are used from 20-30 days to 3-4 months of age at a dose of 10-15 g of active substance per 1 kg of live weight 2 times a day, alternating with drugs ten times a day (2).

As a preferred analogue, we consider a method of therapeutic and prophylactic intervention, including the use of biovit-80, a drug of the antibiotic chlortetracycline with vitamin B ₁₂ (3).

A common disadvantage of the known methods is the low prophylactic effectiveness due to the weakly pronounced effect of the antibiotic on the biochemical processes in healthy animals compared with patients; the appearance of secondary dysbiosis as a result of a violation of intestinal microbiocenosis (quantitative restructuring of the intestinal biocenosis and a sharp change in the qualitative characteristics of its constituent joints) and antibiotic-resistant strains. In addition, the long-term use of tetracycline drugs is accompanied by numerous side effects associated with damage to the mucous membranes, suppression of local immunity and factors of natural resistance.

The purpose of the invention is improving efficiency.

The goal is achieved by using as a tetracycline preparation a composite preparation furacycline M in combination with okarin.

Furacycline M is a therapeutic and prophylactic agent containing an antibacterial composition represented by oxytetracycline hydrochloride, norsulfazole, furagin and a combination of vitamins represented by thiamine bromide and belatin as a source of B vitamins that are not synthesized in the intestines of calves up to 3-4 months of age , but are necessary for normal digestion, regulation of metabolism, in particular protein, stimulation of hematopoiesis, increase the body's resistance to infection. When prescribing antibiotics and sulfonamides, the body's demand for B vitamins sharply increases (4).

Probiotic okarin is a lyophilized biomass of living microbial cultures - representatives of the normal intestinal microflora of a healthy person, including three strains of E. coli and fecal enterococcus. The microbes that make up okarin participate in the formation of parietal microflora of the intestine, are characterized by low metabolic activity and reproduction, which leads to their antibiotic resistance. High adhesiveness suppresses the growth of pathogenic and conditionally pathogenic microflora, promoting the growth of bifidobacteria and lactobacilli (5).

The method is as follows.

Calves from 12 days of age for 10 days are given orally with milk furacycline M at a dose of 0.4-0.5 g / kg live weight 2 times a day and ocarin at a dose of 8 × 10 ⁶ microbial cells in 100 ml of boiled and cooled to a temperature of 38-39 ° C water 1 time per day at night.

Checking the claimed technical solution for compliance with the criterion of “inventive step” showed that neither the scientific nor patent literature revealed a combination of features specified in the characterizing part of the claims. The combined use of the probiotic ocarin and the composite drug furacycline M, the individual components of which have immunosuppressive properties, provides for the correction of key disturbed parts of the immune system, mainly the cellular component of the immune system, as a result of an increase in the functional activity of blood neutrophils, an increase in the content of T and B lymphocytes and activation of their function , which plays a major role in chlamydial infections.

The effectiveness of the prophylaxis method by the combined use of furacycline M and ocarin was determined in 3 farms of the Nizhny Novgorod region with a high level of complement-binding antibodies (from 1:16 to 1: 256) in 50-60% of healthy animals, which indicated the presence of latent infection in these farms. The incidence of chlamydial bronchopneumonia in calves 15-20 days of age was 57-63%. The diagnosis was confirmed by clinical, pathological methods and laboratory studies according to the “Guidelines for the laboratory diagnosis of chlamydial infections in animals”, approved by the Veterinary Department of the Ministry of Agriculture of the Russian Federation, June 30, 1999. The situation was exacerbated by immunodeficiencies that were detected in clinically healthy calves 12-15 days of age ( Table 1).

Table 1 The immune status of calves 12-15 days of age Indicators Clinically healthy calves in a dysfunctional economy Clinically Healthy Calves White blood cells, 10 ⁹ / L 6.3 ± 0.5 6.9 ± 0.3 Lymphocytes,% 45.0 ± 1.5 59.2 ± 1.6 T-lymphocytes,% 18.9 ± 1.2 29.7 ± 1.3 B-lymphocytes,% 7.1 ± 0.4 10.9 ± 0.9 Functional activity of blood neutrophils in the HCT test,% - spontaneous 6.8 ± 0.9 8.2 ± 0.3 - induced 16.8 ± 1.0 25.6 ± 1.2 Bactericidal activity of blood serum,% 36.2 ± 1.8 60.0 ± 1.6 Lysozyme activity of blood serum,% 2.2 ± 0.2 3.2 ± 0.1 β-lysine activity of blood serum,% 18.6 ± 1.4 28.2 ± 1.2

As can be seen from table 1, in clinically healthy calves of a chlamydia-poor economy, there were immune disorders of the cell link, in particular lymphopenia, a decrease in the functional activity of blood neutrophils and the main indicators of natural resistance compared to calves-analogues of a farm that was successful in this disease.

So, the relative content of lymphocytes was lower by 24, T-lymphocytes by 36.4, B-lymphocytes by 34.9%, the functional activity of blood neutrophils in the spontaneous test by 17, in the induced by 34.4%, the bactericidal activity of blood serum 39.9, lysozyme activity by 31.3, β-lysine activity of blood serum by 34%.

The essence of the method is illustrated by examples.

Example 1. To determine the effect of the drug ocarin at a dose of 8 × 10 ⁶ microbial cells with a single daily intake for 10 days on the immune status of calves in a farm that is permanently dysfunctional due to chlamydial bronchopneumonia, 3 groups of 12-day-old calves were formed with 10 animals each each. Calves of all groups were fed in accordance with growing technology. The calves of the 1st and 2nd groups were given once a day for 10 days inside Biovit-80 at a dose of 0.25 g / kg of live weight, the calves of the 1st group were additionally given inside the ocarin at a dose of 8 × 10 ⁶ microbial cells Once a day at night for 10 days. Calves of the 3rd group of drugs were not received.

The calf immune status after a course of prophylaxis was evaluated by the absolute number of leukocytes, the relative content of lymphocytes (determined by generally accepted methods), the relative content of T- and B-lymphocytes (determined by the method of rosette formation), and by the indicators of the functional activity of blood neutrophils (determined in the reaction of nitrosine tetrazolium recovery by A .N. Mayansky et al., 1979), bactericidal activity of blood serum (determined by the method of O.V. Smirnova, T.A. Kuzmina, 1966), lysozyme activity of blood serum (op distributed according to the method of V.G. Dorofeychuk, 1968). By standard bacteriological methods from all calves, fecal samples were examined to determine the state of normobiosis of the intestinal microflora. The research results are presented in tables 2 and 3.

table 2 The immune status of calves after a preventive course Indicators Calf groups first second the third White blood cells, 10 ⁹ / L 8.9 ± 0.2 8.0 ± 0.5 6.9 ± 0.4 Lymphocytes,% 52.0 ± 1.2 50.3 ± 0.9 45.1 ± 0.5 T-lymphocytes,% 25.9 ± 0.4 21.7 ± 0.3 17.0 ± 0.9 B-lymphocytes,% 11.9 ± 0.7 8.5 ± 0.8 7.0 ± 0.4 Functional activity of blood neutrophils (HCT-test),%: - spontaneous 7.8 ± 0.3 7.0 ± 0.3 6.1 ± 0.2 - induced 28.7 ± 1.2 20.5 ± 1.5 15.4 ± 0.5 Bactericidal activity of blood serum,% 43.4 ± 1.4 36.0 ± 1.1 32.1 ± 0.9 Lysozyme activity of blood serum,% 5.0 ± 0.5 3.8 ± 0.3 3.0 ± 0.3

From the data of table 2 it can be seen that the combined use of the drug Biovit-80 and ocarin to calves of the 1st group contributed to a more pronounced increase in the indicators of the immune status compared with the 2nd and 3rd groups. At the same time, the absolute content of leukocytes increased by 11.3 and 29%, the relative content of lymphocytes - by 3.4 and 15.3%, T-lymphocytes - by 19.4 and 52.4%, B-lymphocytes - by 40 and 70 %, respectively, the functional activity of blood neutrophils in a spontaneous test increased by 11.4 and 27.9%, in the induced one by 40 and 86.4%, bactericidal activity by 20.6 and 35.2%, lysozyme activity of blood serum by 31.6 and 66.7%, respectively.

Table 3 The results of microbiological studies of feces of calves 22 days of age Indicators The number of allocated microorganisms, lg / g Calf groups first second the third Resident microflora: Bifidobacteria 10-11 6-7 1-2 Lactobacilli 7-8 3-4 0-1 Esherichia 6-7 6-8 7-9 Enterococci 3-4 4-6 5-7 Bacteroids 2-4 2-5 4-6 Clostridia 1-2 2-3 2-5 Candidates 0-1 1-2 3-4 Transient microflora: Klebsiella 0 0 0-1 Cytobacteria 0-1 1-2 1-3 Enterobacter 1-2 1-2 4-5 Pseudomonads 0 0 0 Protea 0 1-2 2-4

As can be seen from the data in Table 3, the combined use of biovit-80 and ocarin by the calves of the first group of the calves contributed to the normalization of intestinal microbiosis with the establishment of the dominant position of resident microflora and its main physiological significance representatives - bifidobacteria and lactobacilli. The predominance of bifidobacteria and lactobacilli in the intestine provided an immunostimulating effect and colonization resistance, preventing the multiplication of pathogenic and conditionally pathogenic microorganisms, their translocation and the occurrence of endogenous infection. A decrease in the number of bifidobacteria less than 7 lg / g and lactobacilli less than 2 lg / g of intestinal contents of calves of the 2nd and 3rd groups indicated a different degree of disturbance of intestinal microbiocenosis.

Example 2. The comparative effectiveness of the claimed and known methods of prevention was evaluated according to the results of clinical and bacteriological studies of samples of feces and nasopharyngeal mucus of calves.

3 groups of 12-day-old calves were selected, 43 heads each. The calves were fed in accordance with growing technology. Calves of the 1st group (experimental) for 10 days were given orally furacycline M and ocarin according to the scheme: furacycline M at a dose of 0.5 g / kg live weight 2 times a day and ocarin at a dose of 8 × 10 ⁶ microbial cells 1 time per day to night. Calves of the 2nd group (prototype method) were given inside Biovit-80 at a dose of 0.25 g / kg live weight 1 time per day for 10 days. Group 3 calves (the basic method) were injected intramuscularly with oxytetracycline at a dose of 9 mg / kg body weight 2 times a day for 10 days.

Clinical observations were conducted on experimental calves. Prophylactic efficacy was assessed by the number of sick calves, percentage of safety and average daily gain in live weight for the first month of life.

At the end of prophylactic treatments, bacteriological studies of fecal samples were carried out for the detection of bifidobacteria using Blaurock medium and lactobacilli - MPC-4 medium. Nasopharyngeal mucus from live and dead calves was examined for the detection of bacteria, followed by determination of their pathogenicity by generally accepted methods of bacteriological analysis.

The results of clinical studies are presented in table 4.

Table 4 The comparative effectiveness of the claimed and known methods for the prevention of chlamydial bronchopneumonia of calves Indicators Calf groups The first is the claimed method The second is the prototype method Third - the basic way The number of calves subjected to preventive treatment, goal. 43 43 43 Of them fell ill with signs of chlamydial bronchopneumonia, a goal. (%) 2 (4.6) 5 (11.6) 11 (25.6) The calves fell, the goal. (%) 0 1 (2,3) 3 (7) Preventive efficacy,% 95.4 88.4 74,4 Preservation,% 100 97.7 93 The average daily gain in live weight for the 1st month of life, g 517 445 377

As can be seen from the data in table 4, clinical data confirmed the effectiveness of the claimed method for the prevention of chlamydial bronchopneumonia of calves in comparison with the prototype and the basic method, which was expressed in a decrease in incidence of 7 and 21%, respectively, increasing their safety to 100 compared to 97.7 and 93 %, respectively, and average daily weight gain for the first month of life by 16.2 and 37%, respectively.

In bacteriological studies of fecal samples in 22-day-old calves of the 1st group, receiving furacycline M and ocarin inside, the number of bifidobacteria was 9-12 lg / g, lactobacilli - 2-4 lg / g; in calves of the 2nd group, receiving biovit-80 inside, the number of bifidobacteria was 6-7 lg / g, lactobacilli - 1-2 lg / g; in calves of the 3rd group, the number of bifidobacteria was 2-4 lg / g, lactobacilli - 1-2 lg / g.

The results of microbiological studies of nasopharyngeal mucus of calves are presented in table 5.

Table 5 Microbiological indicators of nasopharyngeal mucus of calves Calf groups Isolated microorganisms 1st (claimed method), n = 43 2nd (prototype method), n = 43 3rd (basic method), n = 43 Staphylococci

Streptococcus

Peptococcus

Esherichia

Klebsiella

Pasteurella

Salmonella

Pseudomonads

Total

Note: in the numerator - the number of selected cultures, in the denominator - the number of pathogenic crops.

From the data in Table 5 it is seen that 80 non-pathogenic cultures, represented by 5 genera of microorganisms, mainly gram-positive cocci, were isolated from the upper respiratory tract of calves of group 1, while staphylococci were found in 67.4% of samples, streptococci in 60.5% of samples , peptococci - in 48.8% of samples, Escherichia and Klebsiella - in 4.7% of samples.

From the calves of the 2nd group, 106 cultures of microorganisms, representatives of 7 genera, were isolated, while 13.2% of the cultures were pathogenic. In calves of the 3rd group, the microflora of the upper respiratory tract was represented by 8 genera, and 55 or 32% of the 172 isolated cultures were pathogenic.

The results indicate that, against the background of a decrease in barrier functions and body resistance in calves of the 2nd and especially 3rd groups, the activity of the commensals - representatives of the conditionally pathogenic microflora, the manifestation of its pathogenic effect complicating chlamydial infection.

Studies have confirmed the effectiveness of the combined use of furacycline M and ocarin for chlamydial bronchopneumonia in conditions of prolonged chlamydia carrier, i.e. immunological pressure. The proposed combination can significantly reduce side effects, provides immunocorrection of key broken links of the immune system and increase the body's natural resistance.

Sources of information

1. Chlamydia of farm animals./ N.E. Khazipov, Kh.Z. Gaffarov, R. A. Igafikova and others. - M .: Kolos, 1984. - S. 94-95.

2. Suleymanov S.M. Prevention of respiratory diseases of calves // Veterinary medicine. - 1986, No. 11. - C.11-13.

3. Chlamydia of cattle and measures to combat it. Guidelines. Cheboksary, 2001 .-- P.47.

4. Guidance on the use of furacycline M in veterinary medicine No. 13 / 5-2 / 1850.

5. Kulchitskaya M.A. Modern aspects of probiotic biotechnology // Problems of Veterinary Medicine at the Turn of the Century: Sat. scientific tr NSAA, N. Novgorod, 2001 .-- S.293-296.

Claims (1)

A method for the prevention of calf chlamydial bronchopneumonia, including the use of a tetracycline preparation inside, characterized in that calves from 12 days of age use furacycline M and an additional probiotic ocarin for 10 days according to the following scheme: furacycline M at a dose of 0.4-0 5 g / kg of live weight 2 times a day, ocarin - at a dose of 8 × 10 ⁶ microbial cells 1 time per day at night.