CS200048B1 - Agent for dysentery and growth defects prevention of swine and method of producing the same - Google Patents

Description

(54) Preparation for the prevention of dysentery and growth disorders in pigs and how to produce it

The invention relates to the composition and preparation of a product intended for the prevention of dysentery and growth disorders in pigs during dairy nutrition and in pre-starch, with the working name Lazol.

Pig dysentery is a disease that causes considerable problems in all countries with significant pig breeding. Due to the fact that pig dysentery occurs predominantly in large-scale pigs and mortality and morbidity is high, it is necessary to pay increased attention to it · / Harris D.L./tinyon J.M .: Swine disentery: History, recent discoverles and eondepts. ^ rooeedings 23rd Annual Pfizer Research Conference, Aug 11, 9-25, 1975 »

Urban VP, Šnur VI »World Veterinary Congress Summaries, ⁱⁿ ol. 2, Thessaloniki, Oreece, 1975].

Pig dysentery is most commonly found in the weight category of 15 to 70 kg, but may also have occurred in piglets and fattening pigs. The transmission of the disease is most often due to the ingestion of faeces or transmission from carriers that do not show clinical signs.

Under operating conditions, morbidity is about 90% and mortality, depending on the efficacy of treatment, can be up to 30%. Under experimental conditions, if animals are not treated, the mortality is up to 50% / H _and rris DL, Qlock RD! Swine dysentery. Ins Diseaser swine. Fourth Edition, The Low State University Press, 1975 /.

For the prevention and treatment of pig dysentery, a number of drugs have been proposed to date, which have been used in practice for various periods of time with various interventions.

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Streptomyoin, bacitracin, chloramphenicol, erythremoin, tylosin, gentamyoin, lincomyoin, neomyoin, spiramyoin and virginiamycin have been tested in this indication. In view of the relatively rapid growth of the germ line to these antibiotics, attention has been drawn in various countries to aromatic arsenic compounds / manilic acid, 3-nitro-4-phenylazzonic acid and the like, carbadox and in particular to 5-nitroimidazole derivatives, The most widely used application is currently dimetridazole and ronidazole / e.g. Grifin R.M.t Therapy and prophylaotio aotivity of dimetridazole against experimentally transaitted swine dyeentery, fet.Reo. 91/5 /, 349 to 353.1972; Cottereau F .: Traitement du Syndrome entéritique hémorhagique du pere par la dimetridazole.

Revue de médieine vétérinaire 122, 361-369, 1971 $ Meeeersmith R.S. > Effect of lpronidpzol on awine dyeentery. Vet. BEA. email Anim. Clin. 68/9 /, 1021-1033, 1973 $ Taylor D.J. »Ronidazole β in the treatment of experimental awine dyeentery. * Et. Reo. 95 (10), 215-216, 1974 $ Williams B.J., Shively J.E. In vitro antitreponemal activitiea of carbaddx, virginiamycin, elaquindox and tylosin and indieea of their effeotlvenea for preventing awine dysentery. Vet. Med. email Anim. Clin. 73, 349-351 (1978)].

Ronidazole (1-methyl-2-C (carbamoyloxy) -methyl7-5-nitroimidazole), along with other 5-nitroimidazole derivatives, such as metronidazole, dimetridazole, ipronldazole, omidazole and tinidazole, is one of the modern high-performance chemotherapeutics with extensive antiprotozoer activity. It is also effective against mycoplasmas. In addition to its action against mycoplasma and protozoa, ronidazole also has remarkable antibacterial properties, which have been demonstrated both in vitro and in vivo in the mouse / Miller A. K. experiment. Antibaoterial aotivity of ronidazole. Appl. Microbiol. 22 (3), 480-481, 1971. In vitro, ronidazole is most effective against Treponema hyodysenteriae, Campylobaoter coli and Clostridlum perfringens. The high efficacy of ronldazole against T. hyodysenteriae is explained by the rapid disappearance of the germs from the feces of pigs treated with ronidazole / Taylor D. J. Ronidazole in the treatment and prophylaxis of experimental swine dyeentery. Vet. Reo. 99, 453-456, 1976].

For stimulating the growth of pigs for fattening, ronidazole Box J.R. et al. / Performanoe of piga feed grade4 levels of ronidazole. J. Anim. Sol. 34 (3), 470-472, 1970] and in the treatment of experimental dysentery in 1974 by Taylor D.J. / Ronidazole in the treatment of experiential / mental awine dissenters. Vet.Rea. 95 (10), 215-216. Since then, aa and ronidazole have done a number of studies that aa have not optimized the dosage of the active ingredient in drinking water and feed in experimentally induced burns, but also in clinical conditions. Based on the results achieved, the authors recommend to choose 0.006% concentralaria because a / recurrence of inflammation is minimal, b / the concentrate used can eliminate most spirochet from the column, o / use and less drug than conoentralaria 0.012%, i / conentralaria 0.006% water intake in pigs.

In recent years aa pays considerable attention to the so-called. a microbiotic / problematic / preparation that stabilizes stabilized live germs of lactic acid bacteria, lactobaoil; and streptococci. Feroral application of these constructive timber germs induces primary positive qualitative changes in intestinal bacterial biocoenosis (qualitative decrease in conditionally pathogenic and metabolic strains of destructive strains), which manifested aa secondary to improved general health, increased resistance, improved B activity, and improved growth rate J .: Results of Experiments Using Microbiotics in Livestock Nutrition. Veterinary 26/8 /, 363-367,1976 /.

The main idea of the so-called. lactobaeil therapy is characterized by the fact that lactohaoils and other non-pathogenic intestinal germs of RF species (especially streptococci) affect pathogenic species, mainly E. coli, adversely. The formation of lactic acid does not cause a shift in the pH of the gastrointestinal tract to the acid side, thereby partially inhibiting both growth and toxin production.

A wide variety of effective microbiotic agents are produced abroad for human medicine, which are indicated for the treatment of gastronintestinal disorders and infectious enteritis in infants. In the field of veterinary medicine, the use of microbiotic preparations has been implicated in the tradiol. The first experiments with microbiotics in the therapy and nutrition of animals have been in the mid-sixties, mainly for the control of enteritis, treatment of colibacillosis and stimulation of growth / Kohler E.I., Bohl E. M. Prophylaxis of diarrhea in newborn pigs. J. Am. med. Agg. 144, 1294-1297, 1964 $ Redmond H.E., lloore R.W .: Biological effeot of introducing Lbe. Acidophllus in a large swine herd experiencing enteritia. Seduce. Vet. 18, 287-288,1965 $ King J.O.I. »Laotobacillus as growth stimulants for pigs. Veterinaria, London, 5/4 /, 273-280, 1968 $ Varga, J., Mészáros J .: Kétletek a szopos malaook és borjak bélfórajanak bsfolyásoláaá lactobacilusokat tartalmazó készítményel, Jjagy. Állatorv. Lap. 5,301303, 1974].

A more extensive introduction of microbiotics into veterinary practice has been the recent introduction of modern conservation microbiological techniques, allowing the production of stabilized concentrated microbial cultures. A new technology for the production of microbiotics for veterinary purposes is coming from the Swedish company Medipharm. The basic ingredient of Lactiferm is a stabilized Streptocoocus faecium culture. Significant reductions in the frequency of gastrointestinal disorders, improved average weight gain, improved feed conversion and improved adaptability to new conditions were found in the multiple tests conducted with this formulation. / Mican P. et al .: Use of microbiotics in pig rearing and fattening. Krmivarstvi 8, 180-183, 1976].

A disadvantage of said method of administering microbiotics is that in areas where swine dysentery is present, the stimulatory growth effect of said microbiotic cannot be fully utilized, since it alone is not sufficient for therapy or prevention of dysentery. Therefore, the present invention provides a combination of the Streptocoocus faecium microblot with the chemotherapeutic 1-methyl-2 - [(carbamoyloxy) methyl] -5-nitroimidazole to which this strain is resistant. The possibility of combining Streptooooous faecium with ronidazole in terms of survival and the ability to reproduce Streptococoue faecium was laboratory tested. Ronldazole does not inhibit the embryo-propagation of this strain. The ability to rapidly colonize Streptooooous faecium in the gastrointestinal tract is maintained, while the preventive protection of pigs against dysbnteria and other gastrositritis is maintained. The advantage of the combination of 1-methyl-2- (carbamoyloxy) -methyl] -5-nitroimidazole / ronidazole / β bacterial strain Streptocoocus faeelua in Lazol is the very good preventive protection of pigs against dysenteryl, while providing significant growth inhibition and improved feed conversion.

The Streptocoocus faeoium strain was deposited in the Czechoslovak Culture Collection at the Veterinary Research Institute in Brno under the number CCM 6142.

The preparation according to the invention contains, in 1 g, 12 to 18 mg of 1-methyl-2- ['carbamoyloxy] methyl | -5-nitroimidazole, at least 1.10 live germs of Streptoooccus faeoium and 6 to 10 mg of zinc oxide. The preparation is applied at 0.5% concentration in the appropriate feed mix, well homogenized. It is used as a preventative in starter dairy feeds and in starter continuously or intermittently at 12-day intervals.

Example 1

The primary bacterial culture of the strain Streptoooocus faeoium, CClI 6142, which has been flushed to ronidazole obtained by passage through this chemotherapeutic drug, is propagated on a soy nutrient medium in a thermostate at 37 ° C for 16 hours, forming a rich sediment. prepared sterile modified milk medium in a fermenter, containing 2% by weight of skimmed-milk powder and 0,3% by weight of dried yeast autolysate. The inoculated medium is cultured under aeration and stirring at 37 ° C to 45 ° C for 16-16 hours. The bacterial culture thus prepared, containing 10 µg to 10 µg germs per ml, is fixed by drying (lyophilization, fluidized bed, spray drying).

and increasing the stability and resistance to the other ingredients of the formulation and to the environment, the dried bacterial culture is stabilized by fluidized bed / e.g. on a WSG Glatt fluidized bed granulator (3% aqueous or ethanolic solution of polyvinylpyrrolidone (Kollidon 30)). Fluidization takes place at a temperature of 35 to 40 ° C,

The stabilized bacterial culture of the strain Streptoooccus faecium, containing at least

1.10 live germs (1 g are mixed with ronidazole in such a proportion that the finished product contains 12 mg ronidazole (1-methyl-2-f (carbamoyloxy) methyl) -5-nitroimidazole) in 1 gram *

at least 1.10 live Streptoooocus faeoium germs and 8 mg of zinc oxide. Prior to homogenization, said amount was technologically adjusted and supplemented to 1 g starch-modified microbially safe vehicle (heat and enzymatically treated starch, which is produced under our low Solvarin M 20, Amylia). Sucrose and kaolin may also be used as vehicles.

Example 2 v

The primary bacterial culture of the ronidazole-resistant Streptocoocus faeoium strain is propagated on a soy nutrient medium in a thermostate at 37 ° C for 16 hours. The expanded bacterial culture is seeded with the prepared sterilized modified milk handles in a fermenter. the codified milk medium contains 20% by weight of skimmed milk powder, 0.1% by weight of dried yeast autolysate and 1% ammonium sulfate. The inoculated medium is cultured under aeration and stirring at 37 to 45 ° C for 16 to 16 hours, whereby the prepared bacterial culture is fixed by drying and stabilized by fluid flow as set forth in point 1.

The stabilized bacterial culture of Streptoooocus faecium is mixed with ronidazole and zinc oxide in a ratio such that the finished formulation contains 1 mg of 1-methyl-2-f (carbamoyloxy) methyl-5-nitroimidazole (ronidazole) and 10 mg per gram. oxide »

zinc and at least 1.10 live Streptocoocue faecium. further ea manages as described in Example 1.

Example 3

The primary bacterial culture of the Streptocoocue faeoium strain is propagated on a masopepton broth in a thermostate at 37 ° C for 16 hours, further processed as described in Example 1, except that the stabilized bacterial culture of Steagtooocous faeoium is mixed with ronazonium acid and ronazonium acid. in such a ratio that the finished formulation contained 16 mg of 1-methyl-2G- (caxbamoyloxy) -methyl-3,5-nitroimidazole in 1 gram, had at least 1.10 live Streptooooous faeoium and 6 mg of zinc oxide.

The efficacy of the product for the prevention of dysentery and rhinitis in pigs is documented by the following tables:

Table S. 1: Experiment in the feeding period of the feed mixture COS 2 / Comprehensively for early rearing of starch - starter /.

COS 2 ingredients: fish meal, dried torula, skimmed milk powder, dried potato flakes, soya cake, malt flower, technical sugar, cereal germ, wheat, • barley, husked oats, dicaloium phosphate, feed salt, mineral feed, vitamin antibiotic Appendix.

Experimental Control Group

Number of pigs in group / pcs / 100 100 Average live weight (w / w) per piece at the start of the experiment 17.82 18.96 Number of feeding days 93 93 Death during experiment (pcs) 1 2 Number of pigs per konoi experiment 99 98 Average w./pcs per experiment / kg / 61,51 55.50 Absolute increase in body weight per piece / kg / 43.69 36.54 Average increase in body weight / pc and day / kg / 0,473 0,393 Increase index w / pc / day / /% / 120.0 100.0 Feed consumption per kg bw / Kg / 3.8 4.0 Feed consumption index per kg bw /% / 95.0 100.0

Note: The control group was positive with Ridzol P,

i preparation containing 12% ronidazole. The experiments were carried out on pigs in a dysenteric locality. In the course of the experiment, dysentery did not appear in either group.

Table no. 2: I. Experiment in the period of feed mixture COS 2.

experimental group Controlling group Number of pigs per group 321 321 Average live weight / w / piece per start of experiment / kg / 9.0 7.0 Number of feeding days 18 36 Death during experiment (pcs) 1 0 Number of pigs at the end of the experiment (pcs) 321 321 Average w.h per experiment nonci / kg / 20.90 19.96 Absolute increase in body weight per piece / kg / 11.50 12.96 Mean w.h. per piece per day / kg / 0,673 0,370 Index increase w / w ha piece and day /% / 181.8 100.0 Feed consumption per kg bw / Kg / 1.36 1.44 Feed consumption index per kg bw /% / 94,44 100.00

II. Experiment in the period of compound feed A 1 / Complete mixture for pigs up to 35 kg / · Ingredients: whitewash concentrate, vitamin lucerne meal, bun flakes, diapers, barley,

oat, salt, mineral feed additive. Experimental Control group group Number of pigs in group / pcs / 320 321 Average live weight per head at start of experiment / kg / 20.90 19.96 Number of feeding days 91 92 Death during experiment (pcs) 1 * Necessary slaughter during the experiment (pcs) 1 17 Number of pigs at the end of the experiment (pcs) 318 300 Average w / pc at end of experiment / kg / 63 59 Absolute increase in body weight per piece / kg / 42.10 39.04 Mean w.h. per piece per day / kg / 0,473 0,405 Increase w / w index per piece and day /% / 116.79 100.00 Feed consumption per kg bw / Kg / 2.90 3.70 Feed consumption index per kg bw /% / 76.38 100.00

Note: The control group was negative, with no feed mix medication, but in the case of the occurrence of climacteric dysentery symptoms, it was treated with Vibriogal, i. preparation containing 11% dimetridazole, 6,5% furazolldone and 2% zinc oxide.

TabuJka δ. 3: Experiment in the feed mix A 1 / Complete mixture for pigs up to 35 kg /.

Experimental Checklist

group group Number of pigs in group / pcs / 447 455 Avg. per piece at the beginning of the experiment / kg / 20.91 20.91 Number of feeding days 82 82 Death during experiment (pcs) 7 20 Percent mortality during experiment /% / 1.58 4.39 Number of pigs at the end of the experiment (pcs) 440 ⁴³⁵ Avg. on trial / kg / 70.29 62.60 Absolute increase in body weight + pcs / kg / 49,38 41,69 Mean w.h. per piece per day / kg / 0,519 0,441 Increase w / w index / pcs and day /% / 117.95 100.0 Feed consumption per kg bw / Kg / 3.08 3.21 Feed consumption index per kg bw /% / --....... 100.0

Note: The experimental group was balanced in this experiment and no morbidity was recorded throughout the course. The pigs were lively, with no signs of diarrhea and a reduced cough was also observed in the ointment. The causes of death in this group (1.58%) were mainly bronohopneumonia. No treatment or interventions were conducted in this group. A total of 20 pigs died in the control group. In this group remained salmonelloza, dysentery and diseases of the respiratory system, ^{ώ β} 1léčbu were used 76 kg of Vibriogal, 1.5 1 Červienková serum and 500 mL of Chronloinu, i.e. preparation containing 15% D-chloramphenicol.

Claims (5)

Table δ. 3t Feeding season experiment A 1 / Complete mixture for oceans up to 35 kg /. Experimental Control group group Number of pigs in group / pcs / 447 455 Average y.h. per piece per experiment / kg / 20,91 20,91 Number of feeding days 82 82 Death during the experiment / pcs / 7 20 Percentage of death during the experiment /% / 1,58 4,39 Number of pigs at the end of the experiment / pcs / 440 435 Průměrhá ž.h. per test trial / kg / 70.29 62.60 Absolute Increment + pcs / kg / 49.38 41.69 Average incr. per piece / day / kg / 0.519 0.441 Y.h. / pc and day /% / 117.95 100.0 Feed consumption per kg bw. / kg / 3.08 3.21 Feed consumption index per kg b.h. /% / --- 22a $ ... 100.0 Note The experimental group was balanced in this experiment and no morbidity was observed throughout the course. The pigs were alert, with no signs of diarrhea, and cough was also reduced in the whole group. The causes of death in this group (1.58%) were mainly bronohopneumonia. Neither treatment nor interventions were performed in this group. A total of 20 pigs died in the control group. In this group, Salmonellosis, dysentery and respiratory disease persisted, 76 kg of Vibriogal, 1.5 1 of reddish serum and 500 ml of Chlorloin, a formulation containing 15% of D-chloramphenicol, were used for β-drug. PEE DME I ΪΪΕ LE LE U A preparation for the prevention of dysentery and porcine growth disorders, characterized in that in 1 g it contains 12 to 18 mg of 1-methyl-2 G (carbamoyloxy) methyl-3-5-nitroimidazole, minus 6 to 1.10 live germs microorganisms Streptococcus faecium strain, 6 to 10 mg zinc zinc and residual to 1 g microbially safe vehicle. 2. A composition for the prevention of dysentery and growth disorder according to claim 1, characterized in that the microorganism of the Streptocooous faecium strain is a strain of Streptococcus faeciumC011 6142. 3. Preparation of the preparation according to claim 1, characterized in that the propagated bacterial culture of the Streptococcus faecium strain is cultivated on a milk medium modified with yeast autolysate in an amount of 0.1 to 0.3 weight per cent and ammonium sulfate in an amount of 1 to 3 weight per cent. a temperature of 37 ° C to 45 ° C for 16 to 18 hours and a 15 bacterial culture obtained containing 10 to 10% germs fixed by drying and then mixed with 1-methyl-2 C- (carbamoyloxy) -methyl} -5 -nitroimidazole, zinc oxide and microbially harmless vehicle such that 1 g of the composition contains 12 to 18 mg of 1-methyl-2-carbamoyloxy / methyl-3-5-nitroimidazole, at least 1.10 of the live Streptooooo strain and 6 to 10 mg zinc oxide. 4. Preparation of the composition according to claim Z, characterized in that the milk medium is modified with yeast autolysate in an amount of 0.3% by weight and ammonium sulphate in an amount of mass per cent, based on the weight of the medium. 5. Process according to claim 3, characterized in that the hardened bacterial culture is stabilized by a fluidized bed aqueous or ethanolic solution of polyvinylpyrrolidone at a temperature of 35 to 45 ° C.