Method for prophylaxis of chlamydium bronchopneumonia in calves

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

 

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

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

Chlamydia are characterized in the pathogenesis and clinical manifestation as respiratory, enterity, urogenital, articular, neurological, eye, and observeration expressed diseases affecting all organs and systems. They are acutely, chronically and asymptomatic, and patients for a long time remain hemodynamically.

Chlamydial pneumonia (respiratory disease) occurs more frequently in young animals up to 6 months of age. According to some data for respiratory infections chlamydial pneumonia is recorded in 70% of cases. The disease occurs in the form of entitiy, ill mainly calves with 15-20 days of age and older due to the infection of the respiratory tract and low humoral (celastrales) and cellular immunity. The disease is often complicated by secondary (secondary) infection due to activation and pathogenic manifestations in the action of microorganisms - the inhabitants of the upper respiratory tract or translotsirovannoi from the intestines to the weakening of the barrier functions of the body. When pneumonia occurs in severe fatal to 30-40% and above.

Known methods of treatment and preventive intervention for chlamydia, including the use of tetracycline drugs with humidistatic action, in particular biomicin with chlamydial pneumonia (1).

Use of feed antibiotics of the tetracycline group (biovet, Beaterator) with 20-30 days to 3-4 months of age at a dose of 10-15 g of active substance per 1 kg of body weight 2 times a day, alternating drugs every ten days (2).

As the preferred analogue we consider the way of preventive and curative interventions, including the use of biovita-80 - drug antibiotic chlortetracycline with vitamin B12(3).

A common disadvantage of known methods is low prophylactic efficacy due to weakly pronounced effect of the antibiotic on the biochemical processes in healthy animals compared to patients; the occurrence of secondary dysbacteriosis as a result of violations of intestinal microbiocenosis (quantitative reconstruction of intestinal biocenosis and the abrupt change of qualitative characteristics of the components of his followers) and the antibiotic is esistenti strains. In addition, prolonged use of tetracycline drugs is accompanied by numerous side effects associated with lesions of the mucous membranes, the suppression of the local immune system and natural resistance factors.

The purpose of the invention is the increased efficiency.

This goal is achieved by applying a tetracycline drug composite drug furacilin M in combination with drug okarin.

Furacilin M - treatment-and-prophylactic agent containing composition of antibacterial products, presents oxytetracycline hydrochloride, norsulfazola, furagin, and a combination of vitamins, presents thiamine bromide and Blatina as a source of b vitamins, which are not synthesized in the intestines of calves up to 3-4 months of age, but is necessary for normal digestion, regulation of metabolism, in particular protein, stimulation of hemopoiesis, improve the body's resistance to infection. When prescribing antibiotics and sulfonamides dramatically increases the body's need for b vitamins (4).

Probiotic okarin is a freeze-dried biomass of live microbial cultures - representatives of normal microflora of a healthy person, including three strains of Escherichia coli and fecal Enterococcus. Mick is ordinary, included in the preparation of okarin, participate in the formation of parietal microflora of the intestine, characterized by low metabolic activity and reproduction, making them antibioticassociated. High adhesiveness suppresses the growth of pathogenic and conditionally pathogenic microflora, contributing to the growth of bifido - and lactic acid bacteria (5).

The method is as follows.

Calves from 12 days of age for 10 days to give the inside with milk furacilin M at a dose of 0.4-0.5 g/kg body weight 2 times a day and okarin at the dose of 8×106microbial cells in 100 ml of boiled and cooled to a temperature of 38-39°With water 1 time a day at night.

Verification of the claimed technical solution to meet the criterion of “inventive step” showed that neither in science nor in the patent literature have not revealed the combination of features specified in the characterizing part of the claims. The combined use of probiotic Ocarina and composition of the drug furacilin M, the individual components of which have immunosuppressive properties, corrects the broken key components of the immune system, mainly cellular immunity, resulting in the increase of the functional activity of neutrophils in the blood, increasing the content of T - and b-lymphocytes and aktivizaciya functions which plays a major role in chlamydial infections.

Effective prevention concomitant use of furacilin M and Ocarina defined in terms of 3 farms in the Nizhny Novgorod region with a high level complimentative antibodies (from 1:16 to 1:256) at 50-60% of healthy animals, indicating that the presence of these farms latent infection. 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 tests according to the “Methodological guidelines for laboratory diagnosis of chlamydial infections in animals”approved by the veterinary Department of the Ministry of agriculture of the Russian Federation, 30.06.99, the Situation was exacerbated by immunodeficiency, which was identified in clinically healthy calves 12-15 days of age (table 1).

Table 1
The immune status of calves 12-15 days of age
IndicatorsClinically healthy calves troubled economyClinically healthy calves prosperous economy
Leukocytes, 109/l6,3±0,56,9±0,3
Lymphocytes, %45,0±1,559,2&x000B1; 1,6
T-lymphocytes, %18,9±1,229,7±1,3
B-lymphocytes, %7,1±0,410,9±0,9
The functional activity of blood neutrophils in NBT-test %  
- spontaneous6,8±0,98,2±0,3
- induced16,8±1,025,6±1,2
Bactericidal activity of blood serum, %36,2±1,860,0±1,6
The authors estimated the effect activity of blood serum, %2,2±0,23,2±0,1
β-Lisina activity of blood serum, %18,6±1,428,2±1,2

As can be seen from table 1, in clinically healthy calves troubled by chlamydia economy was marked immune disorders cellular level, in particular lymphopenia, reduction of functional activity of blood neutrophils and the main indicators of natural resistance compared with calves peers prosperous on this disease management.

Thus, the relative content of lymphocytes was lower at 24, T-lymphocytes 36.4, b-lymphocytes 34.9%, the functional activity of blood neutrophils in spontaneous test at 17, in induzirovanny the m 34.4%, bactericidal activity of serum 39,9, the authors estimated the effect activity 31.3, β-Lisina activity of blood serum at 34%.

The essence of the method is illustrated by examples.

Example 1. To determine the effect of drug okarin dose of 8×106microbial cells in a single daily admission within 10 days on the immune status of calves in terms of economy, permanently affected by chlamydial pneumonia, formed 3 groups of calves 12-day age of 10 animals each. Calves in all groups were fed in accordance with the growing technology. Calves 1-St and 2-nd group 1 time per day for 10 days was given inside Biovit-80 at a dose of 0.25 g/kg body weight, calves, 1st addition, the group gave the inside of okarin dose of 8×106microbial cells 1 time a day at night for 10 days. Calves 3rd group of drugs have not been received.

The immune status of calves after a course of prophylaxis was assessed by the absolute number of leukocytes, the relative content of lymphocytes (determined by standard methods), the relative content of T - and b-lymphocytes (determined by the method of rosethorne), functional activity of neutrophils blood was determined in the reduction reaction of nitrocine of tetrazole on Annajanska and others, 1979), and bactericidal activity of blood serum (determined by the method Ows novoi, Casesmini, 1966), lysozyme activity of blood serum (determined by the method Vgetarian, 1968). Common methods of bacteriological analysis of all calves was investigated feces samples to determine the status of normalize intestinal microflora. The results are shown in tables 2 and 3.

Table 2
The immune status of calves after preventive course
IndicatorsGroups of calves
 firstsecondthird
Leukocytes, 109/l8,9±0,28,0±0,56,9±0,4
Lymphocytes, %52,0±1,250,3±0,945,1±0,5
T-lymphocytes, %25,9±0,421,7±0,317,0±0,9
B-lymphocytes, %11,9±0,78,5±0,87,0±0,4
The functional activity of blood neutrophils (NBT - test), %:   
- spontaneous7,8±0,37,0±0,36,1±0,2
induzirovanny the 28,7±1,220,5±1,515,4±0,5
Bactericidal activity of blood serum, %43,4±1,436,0±1,132,1±0,9
The authors estimated the effect activity of blood serum, %5,0±0,53,8±0,33,0±0,3

From table 2 it is seen that the combined use of the drug Biovit-80 and Ocarina calves 1st group contributed to a more pronounced increase of indicators of immune status in comparison with the 2nd and 3rd groups. The absolute content of leukocytes increased by 11.3 and 29%, the relative content of lymphocytes - 3.4 and 15.3%, T-lymphocyte - 19.4 and 52.4%, lymphocytes - 40 and 70%, respectively, the functional activity of blood neutrophils in spontaneous test increased by 11.4 and 27.9%in induced - 40 and 86.4%, bactericidal activity by 20.6 and 35.2%, the authors estimated the effect activity of blood serum by 31.6 and 66.7%, respectively.

Table 3
The results of microbiological testing of faeces of calves 22 days old
IndicatorsThe number of selected microorganisms, lg/g
Groups of calves
firstsecondthird
Resident microflora:
Bifidobacteria10-116-71-2
Lactobacillus7-83-40-1
Escherichia6-76-87-9
Enterococci3-44-65-7
The bacteroids2-42-54-6
Clostridium1-22-32-5
Candida0-11-23-4
Transient microflora:
Klebsiella000-1
Citrobacter0-11-21-3
Enterobacter1-21-24-5
Found000
Proteus01-22-4

As can be seen from table 3, the combined application calves 1st group preparativi-80 and Ocarina help normalize intestinal microbiota with the formation of the dominant position of the resident microflora and major physiological significance of its representatives - bifidobacteria and lactobacilli. The predominance of bifidobacteria and lactobacilli in the gut provided immunostimulating effect and colonization resistance, preventing the reproduction of pathogenic and conditionally pathogenic microorganisms, their translocation and the emergence of endogenous infection. The reduction in the number of bifidobacteria less than 7 lg/g and lactobacilli less than 2 lg/g of intestinal content calves 2nd and 3rd groups indicated different degrees of disorders of the intestinal microbiocenosis.

Example 2. Comparative effectiveness declared and known methods of prevention was assessed by the results of clinical and bacteriological studies of samples of faeces and nasopharyngeal mucus calves.

Selected 3 groups of calves 12-day age 43 heads in each. Calves were fed in accordance with the growing technology. Calves 1-th group (experienced) within 10 days gave into furacilin M and okarin scheme: furacilin M at a dose of 0.5 g/kg body weight 2 times a day and okarin dose of 8×106microbial cells 1 time a day at night. The calves of the 2nd group (prototype method) gave inside Biovit-80 at a dose of 0.25 g/kg body weight of 1 times a day for 10 days. Calves 3 groups (basic method) was administered intramuscularly oxytetracycline at a dose of 9 mg/kg body weight 2 times a day for 10 days.

For the experimental calves were clinical observations. PR is a preventive efficacy was assessed by the number of diseased calves, the percentage preservation and average daily gain in live weight during the first month of life.

Upon completion of maintenance treatments conducted bacteriological examination of faeces samples and the detection of bifidobacteria using environment Blaurock and lactobacilli environment Mrs-4. Nasopharyngeal mucus from the living and the dead calves examined for detection of bacteria with subsequent determination of their pathogenicity observatii methods of bacteriological analysis.

Results from clinical trials are presented in table 4.

Table 4
Comparative effectiveness declared and known ways to prevent chlamydial pneumonia of calves
IndicatorsGroups of calves
 The first is the claimed methodThe second method-prototypeThe third basic method
The number of calves subjected to preventive treatment goal.434343
One of them fell ill with signs of chlamydial pneumonia, a goal. (%)2(4,6)5(11,6)11(25,6)
Palo calves goal. (%)0 3(7)
Prophylactic efficacy, %95,488,474,4
Safety, %10097,793
Average daily gain in live weight for the 1st month of life, g517445377

As can be seen from table 4, the clinical data have confirmed the effectiveness of the inventive method for the prevention of chlamydial pneumonia of calves in comparison with the prototype and the basic method, which was reflected in the reduction of morbidity on 7 and 21%, respectively, increasing their safety to 100 in comparison with 97,7 and 93%, respectively, and average daily weight gain during the first month of life by 16.2% and 37%, respectively.

Bacteriological studies of samples of faeces from 22-day-old calves 1st group receiving inside furacilin M and okarin, the number of bifidobacteria was 9-12 lg/g, Lactobacillus - 2-4 lg/g; in calves 2nd group receiving inside Biovit-80, the number of bifidobacteria was 6-7 lg/g, Lactobacillus - 1-2 lg/g; in calves 3rd group the number of bifidobacteria was 2-4 lg/g, Lactobacillus - 1-2 lg/g

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

Table is 5
Microbiological indicators of nasopharyngeal mucus calves
 Groups of calves
The selected microorganisms1-I (the claimed method), n=432-I (prototype method), n=433-I (basic method), n=43
Staphylococcus
Streptococcus
Peptococci
Escherichia
Klebsiella
Pasteurella
Salmonella
Found
Only

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

From table 5 it is seen that from the upper respiratory tract of calves 1 and isolated group of 80 non-pathogenic cultures represented by 5 genera of microorganisms, mainly gram-positive cocci, with staphylococci found in 67.4% of samples, Streptococcus spp - 60.5% of the samples, peptococci - 48,8% of the samples, Escherichia and Klebsiella - 4.7% of the samples.

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

The obtained results indicate that the decline of barrier function and resistance in calves the 2nd and especially the 3rd g the SCP causes the activation of Komissarov - representatives of conditionally pathogenic microflora, manifestations of its pathogenic effects, complicating chlamydial infection.

The conducted research confirmed the effectiveness of the combined use of furacilin M and Ocarina with chlamydial pneumonia in long-term hemodynamically, i.e. immunological pressure. The proposed combination can significantly reduce side effects, provides immunotherapy key impaired immunity and improving the natural resistance of the organism.

Sources of information

1. Chlamydia farm animals./ Nahashon, Hsheppar, Raagapella and others - M.: Kolos, 1984. - S-95.

2. Suleymanov S.M. Prevention of respiratory disease of calves//veterinary medicine. in 1986, No. 11. - C.11-13.

3. Chlamydia cattle and measures against it. Methodical recommendations. Cheboksary, 2001. P.47.

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

5. Kulczycka M.A. Modern aspects of biotechnology probiotics//problems of veterinary medicine at the turn of the century: Sat. the scientific. Tr. MSHA, N. Novgorod, 2001. - S.293-296.

The way to prevent chlamydial pneumonia of calves, including tetracycline oral use of the drug, characterized in that the calves from 12 days of age as tetracycline drug use is furacilin M and advanced probiotic okarin within 10 days according to the following scheme: furacilin M at a dose of 0.4-0.5 g/kg body weight 2 times a day, okarin at the dose of 8×106microbial cells 1 time a day at night.



 

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