RU2404756C1 - Agent and method for treatment of endometritis in cows - Google Patents

Abstract

FIELD: medicine, veterinary science.

SUBSTANCE: invention relates to the field of veterinary science, in particular to methods and agents for treatment of obstetric-gynaecological diseases in cows. Agent contains pefloxacin, metronidazole and chlorhexidine bigluconate, and the base is represented - water-soluble organosilicic derivative of glycerin, having transmucosal activity, composition of which complies with the formula (CH₃)₂Si(C₃H₇O₃)₂·C₃H₈O₃, at the following ratio of components in the agent, wt %: pefloxacin 0.75-1.0; metronidazole 1.0-1.5; chlorhexidine bigluconate 0.05-0.1; water-soluble organosilicic compound - glycerin derivative - up to 100. According to proposed method, introduction of specified agent is carried out in process of complex therapy intrauterinely 3-5 times, with interval of 48 hours, in amount of 50-100 g per single introduction with the help of Janet's syringe and polystyrene pipette for artificial insemination, at the same time prior to introduction the agent is dissolved with sterile distilled water in the ratio of 1:1.

EFFECT: increased efficiency of endometritis treatment in cows.

2 cl, 8 tbl

Description

The invention relates to the field of veterinary medicine, in particular to methods and means of treating obstetric and gynecological diseases in cows, and can be used both in large livestock complexes, and in farm and personal farms.

The well-known "Antibacterial drug" Dorin "(options), a method for the treatment of mastitis and a method for treating endometritis in animals with its use", in which diseased cows are administered intravenously with a frequency of 1 time per day, including rifamycin, doxycycline mixed with a pharmacologically acceptable solvent, at a dose of 0.2-0.4 mg per 1 kg of weight with a course of 1-2 administration (see RF Patent No. 2142804, A61K 31/65, publ. 12/20/99, Bull. No. 35).

With good results of treatment with this drug, the disadvantage of this method of treatment is the use of potent antibiotics in the preparation, which can lead to culling of milk and loss of part of the product, in addition, this form of an aqueous solution makes it difficult for the active ingredients to penetrate the animal tissue.

Also known is the drug enrocid, including enrofloxacin, dimethyl sulfoxide and distilled water, used for the treatment and prevention of postpartum, gynecological diseases in cows and sows. Enrocid is used intrauterine with a Janet syringe to cows in a dose of 100 ml no more than 5 times, after 48 hours in combination with other drugs (see the instruction for the use of the drug Enrocid ZAO NPP Agrofarm. Voronezh, 2003 No. PVR-2- 0.3 \ 01215).

The disadvantages of the drug include its active mono-composition, which can limit its spectrum of activity in the treatment of animals, while dimethyl sulfoxide, which increases the absorption of the drug, has some toxicity, an unpleasant odor and can cause allergic reactions in animals and staff.

The objective of the invention is to develop a broad spectrum of activity for intrauterine administration and a method for its use for the treatment of endometritis in cows, which has not only increased penetration into animal tissue during intrauterine administration, does not cause allergic reactions in animals and staff, but also with increased adhesion to animal tissues, which ensures the prolonged action of the drug in the course of complex therapy with high quality treatment and low manufacturing costs, etc. Menen means.

The problem is solved by the fact that a method and method for treating acute and chronic endometritis in cows has been proposed and clinically tested in laboratory and industrial conditions, using a new antimicrobial agent containing pefloxacin, metronidazole and chlorhexidine bigluconate for the implementation of the method in the course of complex therapy. the basis of the drug used is a water-soluble organosilicon derivative of glycerol with transmucosal activity, the composition of which is em formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · _{3 3 8} About ₃ , with the ratio of components in the tool, wt.%:

pefloxacin - 0.75 ÷ 1.0 metronidazole - 1,0 ÷ 1,5 chlorhexidine bigluconate - 0.05 ÷ 0.1 water-soluble organosilicon compound - derivative of glycerol - up to 100.

According to the proposed method, the administration of the drug is carried out intrauterine 3-5 times, with an interval of 48 hours, at the rate of 50-100 g per injection with a Janet syringe and a polystyrene pipette for artificial insemination, while the drug is diluted with sterile distilled water before administration in a ratio of 1: 1.

The dynamic viscosity of a water-soluble organosilicon derivative of glycerol, (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ O ₃ , is 18.0 Pa · sec (20 ± 0.5 ° C), the determination was carried out on Viscotester 6 rotational viscometer. The kinematic viscosity of a 50% solution (composition) is 8.88 centistokes (mm ² / s), was determined on a Ubbelode viscometer with a capillary diameter of 0.86 mm (determination error ± 1%).

The ratio of the main active components of the drug was selected empirically on the basis of an analytical review and the production experience of the authors, as contributing to a stable, non-stratified composition, convenient for intrauterine administration with a sufficient shelf life and practically proven high therapeutic effect.

The choice of a set of antimicrobials is based on the suppression of the widest possible spectrum of pathogens, including aerobic gram-positive bacteria, aerobic gram-negative bacteria, anaerobic bacteria (spore-forming and non-spore-forming), and protozoa.

Chlorhexidine bigluconate - 1,6-di- (para-chlorophenylguanido) -hexane - is one of the most active local antiseptic agents. It has a fast and strong bactericidal effect on gram-positive and gram-negative bacteria. Depending on the concentration used, it exhibits both bacteriostatic and bactericidal effects with respect to gram-positive and gram-negative bacteria. Bacteriostatic effect is manifested in a concentration of 0.01% or less; bactericidal - in a concentration of more than 0.01%. Fungicidal action - at a concentration of 0.05%. The virucidal effect is manifested at a concentration of 0.01-1.0% (www.remedium.ru/drugs/).

In the proposed tool as a hydrophilic base, a water-soluble organosilicon derivative of glycerol is used, the composition of which corresponds to the formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ O ₃ , which has a set of positive properties: high transmucosal and decongestant activity, favorably affects the processes of epithelization, non-toxic, has optimal viscosity and adhesive properties.

The compound is a clear, colorless, viscous liquid, soluble in water, alcohol, insoluble in chloroform and ether. Obtained by the interaction of dimethyldiethoxysilane with glycerol in a molar ratio of 1.00: 2.65 by heating the reaction mass to 120-130 ° C, holding at this temperature for at least 4 hours with vigorous stirring, followed by removal of the resulting ethyl alcohol.

The compound was developed at the Institute of Organic Synthesis. I.Ya. Pestovsky, Ural Branch of the Russian Academy of Sciences (see Shadrina E.V., Khonina T.G., Boyko A.A. et al. Synthesis and study of water-soluble silicon polyolates and hydrogels based on them. Proceedings of the XI Youth Conference in organic chemistry. Ekaterinburg, 2008 - S.221-224).

A water-soluble organosilicon compound derived from glycerol, (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ O ₃ , has a positive effect on the morphofunctional state of the affected tissues, contributing to reparative processes and improving tissue trophism.

Currently, in the scientific, technical and patent literature, an intrauterine antimicrobial agent is not known that contains, as an active conductive base, a water-soluble organosilicon derivative of glycerol, the composition of which corresponds to the formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ About ₃ ; and also characterized by the totality of the proposed components in the claimed limits of their content.

The proposed comprehensive tool has a wide spectrum of antimicrobial activity and a high ability to stimulate the regeneration of damaged and damaged tissues during the implementation of the proposed method of treatment.

The effectiveness of the proposed tool is due to the presence of three active components of a wide spectrum of activity: pefloxacin, metronidazole, chlorhexidine bigluconate and a biologically active base - a water-soluble organosilicon derivative of glycerol, the composition of which corresponds to the formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ O ₃ .

Used in the proposed method, the tool includes as active ingredients pefloxacin, metronidazole, chlorhexidine bigluconate, which are commercial drugs in the treatment of various bacterial infections.

The preparation of the complex means is carried out by sequential mechanical mixing of the components.

After conducting numerous laboratory experiments and animal studies, temporary regulations and instructions for conducting production tests were developed and approved.

The implementation of the method with the proposed tool based on a water-soluble organosilicon compound - a derivative of glycerol, the composition of which corresponds to the formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · ₃ H ₈ O ₃ , in the form of a convenient form for intrauterine use provides a prolonged the action of the active drug components that make up the product, along with the positive effect of the essential element of silicon, which is part of the product and contributes to the improvement of the reparative processes in the uterine tissues. Moreover, the transmucosal activity of the base allows not only to use small concentrations of active components in the agent, but also leads to their deeper penetration into the affected uterine tissue, accelerating healing. All this in general increases the effectiveness of the drug and accelerate the healing process when implementing the method of treatment, showing the non-obvious effectiveness of the proposed method.

The studies conducted by the authors showed good compatibility of the active components and the base - a water-soluble organosilicon derivative of glycerol, the composition of which corresponds to the formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ O ₃ , and allowed us to identify their limits quantitative content in the product, providing the manifestation of the maximum therapeutic effect. So, when the content of pefloxacin is less than 0.5 wt.% And metronidazole is less than 0.5 wt.%, And chlorhexidine bigluconate is less than 0.01 wt.% (Chlorhexidine bigluconate in the form of a 0.05-0.2% solution) there is a slowdown in the manifestation of a therapeutic effect. The increase in the content of the compounds of a number of fluoroquinolones more than 1.0 wt.% And metronidazole more than 1.0 wt.%, Chlorhexidine bigluconate more than 0.05 wt.% (Chlorhexidine bigluconate in the form of a 0.2% solution), it is impractical since in this case, the occurrence of side effects associated with a violation of the morphofunctional state of the tissues.

Comprehensive production tests of the means and method of administration in obstetric and gynecological diseases of cows were carried out in agricultural organizations of the Sverdlovsk region.

The first scientific and production experiment was carried out at the dairy complex of the Uralets farm of the Ural State Agricultural Academy. Based on the results of a gynecological examination, two groups of cows with postpartum endometritis, 22 heads each, were formed on the basis of analogues.

The treatment regimen for cows is shown in Table 1. The basic therapy included intramuscular injection of a solution of synestrol (factory of Farmadon LLC), oxytocin (ZAO Mosagrogen) and tetramag (ZAO Mosagrogen).

In addition to the basic treatment regimen (basic therapy), the cows of the control group received intrauterine administration of the antimicrobial drug endometrol (manufactured by UrNIVI). The cows of the experimental group were injected into the uterine cavity with the proposed agent, an antimicrobial composition based on a water-soluble organosilicon derivative of glycerol.

Table 1 Treatment regimens for cows in acute endometritis A drug Route of administration Dose Course days Basic therapy Sinestrol Solution 2% intramuscularly 2 ml 12 Oxytocin intramuscularly 40 units 2, 3, 4, 5 Tetramag intramuscularly 6 ml 1.8 Control group (optional) Endometrol into the uterine cavity 100 ml 2, 4, 6, 8 Experienced group (optional) A tool based on a water-soluble organosilicon derivative of glycerol (aqueous solution in a ratio of 1: 1) into the uterine cavity 100 ml 2, 4, 6

Animals of both groups were constantly monitored, taking into account the frequency of drug administration, the duration of treatment, and therapeutic efficacy. After the course, a repeated gynecological examination was performed. While maintaining the clinical signs of the disease, an additional administration of antimicrobials was prescribed. Along with clinical results, immunological parameters were used to control the results. To assess the immune status, a comparison group was used, which included 12 goals of clinically healthy animals. Immunological studies were carried out on the basis of the laboratory of clinical immunology of the Municipal Clinical and Diagnostic Center according to standard methods.

Gynecological examination of cows before treatment indicated a typical course of the disease. In animals, liquefaction was diluted, and the liquefaction sites acquired a grayish color, then the secretion of exudate increased, it had a mucopurulent character, odorless. An increase in total body temperature to 39.6-39.8 ° С was noted in 13.63% of animals, in other cows, body temperature was 39.0-39.5 ° С. During a vaginal examination, hyperemia and swelling of the vaginal mucosa and the vaginal part of the cervix, exudate were detected. During rectal examination, the uterus was enlarged, hung in the abdominal cavity, did not encircle the hand, its walls were thickened, contractility was weak or not detected.

The results of studying the immune status of cows before treatment are shown in Table 2. It was found that in patients with postpartum endometritis, their blood leukocytes were significantly increased in serum, C-reactive protein and the lysosomal-cationic test was reduced. These data are consistent with the results of a clinical study and confirm the presence of an inflammatory process in animals. In addition, in cows with postpartum purulent-catarrhal endometritis, an increased concentration of immune complexes in the serum is observed, which can serve as one of the integral indicators of their participation in the formation of the pathological process.

The state of the phagocytic immunity in cows was evaluated according to the spontaneous and induced HCT test (nitrosine tetrazolium restoration test), which allows to identify components that are formed only during phagocyte stimulation, and therefore it makes it possible to differentiate resting and activated cells. In addition, the NBT test reflects the final reaction of one of the key enzyme cascades responsible for the effector potential of phagocytes, impaired ability to restore the NBT coincides with the pathology of oxygen-dependent mechanisms of bactericidal activity; in this connection, the stimulated NBT test is considered as a cytochemical criterion of readiness for hanging phagocytosis.

The spontaneous HCT test in cows with postpartum purulent-catarrhal endometritis was reduced 1.47-1.56 times compared with clinically healthy animals. In addition, in the stimulated HCT test, the response was low, that is, the reserve of the functional activity of peripheral neutrophils was disrupted. The results of studying the immune status of cows with acute endometritis allow us to conclude that there are significant changes in the state of the immune system. There were no significant differences between the animals of the control and experimental groups in the initial immunological indices.

table 2 The results of a study of the immune status of animals with postpartum purulent-catarrhal endometritis (baseline data) Indicator Comparison group
n = 12 Control group
n = 6 Experienced group
n = 6 White blood cells, 10 ⁹ / l 6.82 ± 0.34 9.81 ± 0.82 * 9.47 ± 1.24 * Lymphocytes, 10 ⁹ / L 3.82 ± 0.28 4.43 ± 0.44 4.12 ± 0.56 CEC, cu 48.6 ± 4.2 115.2 ± 14.9 * 112.2 ± 12.8 * HCT test spontaneous,% 19.85 ± 1.88 13.52 ± 1.98 * 12.70 ± 2.54 * HCT test induced,% 34.67 ± 2.63 12.51 ± 2.78 * 11.24 ± 2.56 * Lysosomal cationic test, cu 2.82 ± 0.02 1.22 ± 0.02 * 1.22 ± 0.02 * C-reactive protein 6.37 ± 0.4 58.62 ± 3.46 ** 64.24 ± 3.10 ** * - the difference with the comparison group is significant, P <0.05; ** - the difference with the comparison group is significant, P <0.01

The course of treatment for cows with acute postpartum endometritis was 8–9 days. After the course, a repeated gynecological examination was performed. While maintaining the clinical signs of the disease, intrauterine administration of antimicrobials continued.

When studying immunological parameters 14 days after the start of treatment, a tendency was found to normalize them in both groups of animals, however, the level of leukocytes, serum concentration of immune complexes and C-reactive protein in the group of control animals remained significantly higher by this time (table 3).

Table 3 The results of the study of the immune status of animals with postpartum purulent-catarrhal endometritis after treatment Indicator Control group n = 6 Experimental group n = 6 White blood cells, 10 ⁹ / l 8.47 ± 0.86 * 5.81 ± 0.43 Lymphocytes, 10 ⁹ / L 2.68 ± 0.44 2.42 ± 0.43 CEC, cu 80.6 ± 10.9 * 51.0 ± 8.8 HCT test spontaneous,% 16.54 ± 3.78 17.85 ± 2.12 HCT test induced,% 22.54 ± 2.06 24.74 ± 4.48 Lysosomal cationic test, cu 2.24 ± 0.03 2.56 ± 0.03 C-reactive protein 22.58 ± 2.42 * 14.2 ± 2.12 * - the difference with control is significant, P <0.05

The treatment period for the cows of the experimental group, which in addition to the basic scheme used the proposed tool - an antimicrobial composition based on a water-soluble organosilicon derivative of glycerol, was shorter by 5.82 days compared to control animals. The effectiveness of treatment, which was understood as recovery with a complete restoration of reproductive ability, in the experimental group was higher by 9.1%.

The duration of treatment and its effectiveness are shown in table 4. When studying the dynamics of clinical indicators, it was found that by the 11-12th day of treatment, the cows of the experimental group stopped exudating, while a transrectal study revealed that the uterus is in the cranial part of the pelvic cavity, it is well reduced on palpation, has an elastic-elastic consistency. At the same time, in cows of the control group, the dynamics of these clinical signs was slowed down. In 40.9% of cases, the control animals were prescribed additional administration of an antimicrobial agent.

Table 4 Therapeutic efficacy of a drug based on a water-soluble organosilicon derivative of glycerol in postpartum endometritis in cows Indicator Control group n = 22 Experimental group n = 22 Duration of treatment, days 17.67 ± 2.3 11.85 ± 1.8 * The effectiveness of treatment,% 86.36 95.45 * - the difference with control is significant, P <0.05

After recovery of the experimental animals and restoration of their sexual cycle, insemination was carried out using deeply frozen sperm in a cervical manner with rectal fixation of the cervix.

Quantitative indicators of the reproductive function of cows are shown in table 5.

Table 5 Indicators of the reproductive function of cows Indicator Control group n = 19 Experimental group n = 21 Restoration of the sexual cycle after childbirth, days 56.64 ± 2.2 49.85 ± 2.8 * The period from birth to fertilization, days 102.5 ± 7.3 80.5 ± 3.1 * Fertility from the first insemination,% 40.90 47.62 * - the difference with control is significant, P <0.05

As can be seen from the presented research results, the use of the proposed method of treatment using means based on a water-soluble organosilicon derivative of glycerol for postpartum endometritis in cows positively affects reproductive function, while the main indicator of reproduction regularity is significantly reduced - the period from childbirth to fertilization.

In the second scientific and production experiment in the farm “Success” on the dairy complex, the effectiveness of an antimicrobial agent based on a water-soluble organosilicon derivative of glycerol in the complex treatment of cows with chronic endometritis was studied. The total number of animals included in the study was 36 animals. The treatment regimen for cows is shown in table 6.

Table 6 Treatment regimens for cows with chronic endometritis A drug Route of administration Dose Course days Basic therapy Sinestrol Solution 2% intramuscularly 2 ml 12 Oxytocin intramuscularly 40 units 2, 3, 4, 5 Tetramag intramuscularly 6 ml eighteen Control group (optional) Endometrol into the uterine cavity 50 ml 2, 4, 6, 8, 10 Experienced group (optional) A tool based on a water-soluble organosilicon derivative of glycerol (aqueous solution in a ratio of 1: 1) into the uterine cavity 50 ml 2, 4, 6, 8

In the preparatory period of the experiment, the experimental and control groups of 18 analog cows with chronic purulent-catarrhal endometritis were formed.

The results of a gynecological study show that the disease in cows was manifested by periodic, less often by the constant release of exudate from the genitals. By its nature, the exudate was mucous, cloudy with an admixture of flakes and streaks of pus, its excretion increased during lying, and also after massage of the uterus through the rectum. The mucous membrane of the vagina and vaginal part of the neck is stagnantly hyperemic, sometimes with point hemorrhages. The cervix is ajar, exudate was revealed in its folds and at the bottom of the vagina. During rectal examination, the uterine horns, as a rule, were lowered into the abdominal cavity, increased in volume by 1.5-2.0 times, contractile function was weak or absent, the walls were thickened. The term from birth to treatment on average for all animals was 50.1 days.

The results of determining the therapeutic effectiveness of the method using an antimicrobial agent based on a water-soluble organosilicon derivative of glycerol for chronic endometritis in cows are shown in table 7.

The data in the table shows that the treatment period for cows in the experimental group was shorter by 7.1 days compared with control animals. The treatment effectiveness, which we estimated to restore reproductive function, was higher in the experimental group by 5.54% compared with the control.

Table 7 Therapeutic efficacy of a drug based on a water-soluble organosilicon derivative of glycerol in chronic purulent-catarrhal endometritis in cows Indicator Control group n = 18 Experimental group n = 18 Duration of treatment, days 18.8 ± 2.1 11.7 ± 1.9 * The effectiveness of treatment,% 83.33 88.89 * - the difference with control is significant, P <0.05

When analyzing the indicators of reproductive function, presented in table 8, it was noted that fertility after two inseminations was higher in the experimental group, where it was 76.47%, while in the control group it was only 53.33%. The period from childbirth to fertilization in the experimental group was shorter by 29.52 days than in the control group.

Table 8 Indicators of the reproductive function of cows Indicator Control group n = 15 Experimental group n-16 The period from the start of treatment to the restoration of sexual cycles, days 41.72 ± 5.16 34.14 ± 4.18 Fertility from the first insemination,% 26.67 31.25 Fertility from the second insemination,% 33.33 37.50 Insemination index 3.24 ± 0.56 2.25 ± 0.48 The period from birth to fertilization, days 129.65 ± 13.89 104.44 ± 7.20 * * - the difference with control is significant, P <0.05

Thus, on the basis of industrial studies, it was concluded that the application of the proposed method using an antimicrobial agent based on a water-soluble organosilicon derivative of glycerol in the complex treatment of cows with acute and chronic endometritis is effective and can be recommended for widespread use in veterinary medicine.

Claims (2)

1. An agent for treating endometritis in cows, including antimicrobial active ingredients and a base, characterized in that the active substances include pefloxacin, metronidazole and chlorhexidine bigluconate as active ingredients, and the base is a water-soluble organosilicon compound - a glycerol derivative with transmucous activity the composition of which corresponds to the formula (CH ₃ ) ₂ Si (C ₃ H ₇ O ₃ ) ₂ · C ₃ H ₈ O ₃ , with a ratio of components in the agent, wt.%:
pefloxacin 0.75-1.0 metronidazole 1.0-1.5 chlorhexidine bigluconate 0.05-0.1 water soluble organosilicon glycerol derivative compound up to 100. 2. A method of treating endometritis in cows, comprising conducting complex therapy with intrauterine administration of antimicrobial agents, characterized in that in the process of treating intrauterine cows, the complex agent according to claim 1 is administered. with increased penetrating effect, moreover, the agent is administered intrauterine 3-5 times, with an interval of 48 hours, at the rate of 50-100 g per administration, while the agent is diluted with sterile distilled water in a ratio of 1: 1 before use.