RU2218924C2 - Method for prophylaxis of diarrhea disease in newborn calves and piglets - Google Patents

Abstract

FIELD: agriculture, veterinary microbiology. SUBSTANCE: method involves prescription of suspension of strains Bacillus subtilis TNP-3-DEP and Bacillus subtilis TNP-5-DEP taken in equal amounts. Method prevents the development of diarrhea and normalizes intestinal microbiocenosis. EFFECT: enhanced effectiveness of method.

Description

 The invention relates to the field of veterinary microbiology and epizootology, in particular to methods for preventing diarrhea of calves and piglets.

 Analogs of the invention. In the literature there are reports of the use of strains of you. subtilis for the prevention and treatment of dysbacteriosis in calves (Topchiy M.P. Application of drugs from live cultures of hay bacillus in dysbacteriosis in calves. - Abstract of thesis. Candidate of Biological Sciences. Minsk, 1979, 21 pp.), increase the gain in pig production ( USSR 634729, class A 23 K 1/00, published December 27, 78), stimulation of nonspecific reactivity (Reznik S.R., Smirnov V.V., Volynets A.K., Vyunitskaya V.A. Effect of oral administration bacteria of the genus Vas. subtilis on the phagocytic activity of blood leukocytes. - Microbiological Journal, 1982, v. 44, 4, pp. 51-54).

 Known bacterial strain "Bacillus subtilis TNP-3-DEPT" used for disinfection of bird droppings and manure from pathogenic microorganisms (patent of the Russian Federation 2105810, C 12 N 1/20, 1998, application 94104662/13 of 03/11/1996, however, this the strain has not been previously used as a drug for preventing diarrheal diseases in animals.

 The basis of the invention is the task of developing an effective environmentally friendly drug for the prevention of diarrheal diseases of newborn calves and piglets.

 For reliable prevention of diarrhea of newborn calves and piglets, a microbial preparation is used, which is a suspension of the bacterial strains Bacillus subtilis TNP-3-DEPT and Bacillus subtilis TNP-5-DEPT. A suspension of bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT" is given individually to calves immediately after birth orally 1 time per day for 3 days at the rate of 1 single dose (10 ml) per head. For therapeutic purposes, the drug is prescribed in the same doses, 3 times a day until the clinical signs of the disease disappear. For piglets, the dose is 5 ml per head. The drug has a wide spectrum of antagonistic activity against pathogenic and conditionally pathogenic microorganisms, pathogens of intestinal and coccal infections. It normalizes intestinal microbiocenosis and increases the body's immunobiological reactivity.

 Morphological signs of strains. In smears of cultures prepared with BCH and MPA, stained by Gram, are gram-positive sticks and short chains (2-3 sticks). Form spores of an oval shape. Capsules do not form.

The cultural properties of the strains. The strains grow well in the BCH at 37 ^o C, form a film on the surface, which when shaken, sinks to the bottom. Broth with slight opalescence. On the MPA and potato agar, the colonies are rounded gray-white. On MPPB they form a weak gray-white precipitate with a slight clouding of the medium. They grow in milk with the addition of 2%, 4%, 6% NaCl, ferment glucose, maltose, levulose without the formation of gas. Indole and hydrogen sulfide do not produce.

 Virulent properties of strains. Strains do not cause the death of rabbits, guinea pigs and white mice by subcutaneous and intraperitoneal administration in doses of 10 million, 5 million and 1 million cells, respectively. Not pathogenic to animals and plants.

 The bacterial strains "Bacillus subtilis TNP-3-DEPT" and "Bacillus subtilis TNP-5-DEPT" differ in antagonistic activity against pathogenic microorganisms.

 The suspension is made from strains of bacteria "Bacillus subtilis TNP-3-DEPT" and "Bacillus subtilis TNP-5-DEPT", deposited in the collection of microorganisms of the All-Russian State Research Institute for Control, Standardization and Certification of Veterinary Medicines (VGNKI) (certificates of deposit of strains dated February 6, 2001, 180/15 and 181/15).

 Patent 2105810 for the invention “Bacillus subtilis bacterial strain used for disinfecting bird droppings and manure from pathogenic microorganisms” was obtained on February 27, 1998 (application 96104662 of March 11, 1996).

 Strains can be obtained in the laboratory for the development of microbial preparations of the Yakutsk Research Institute of Agriculture SB RAAS and VGNKI.

 To prevent diarrhea of newborn calves, a suspension of bacterial strains “Bacillus subtilis TNP-3-DEP and TNP-5-DEP” is administered to calves immediately after birth orally 1 time per day for 3 days at the rate of 1 single dose (10 ml) per head. For therapeutic purposes, the drug is prescribed in the same doses, 3 times a day until the clinical signs of the disease disappear. For piglets, the dose is 5 ml, the drug is used according to the same scheme. Laboratory and production experiments established the ability of the bacterial strains "Bacillus subtilis TNP-3-DEP and TNP-5-DEP" to suppress the viability of pathogens of intestinal and coccal infections, to normalize intestinal microbiocenosis.

The bacterial strains "Bacillus subtilis TNP-3-DEP and TNP-5-DEP" are grown in meat and peptone agar pH 7.0-7.2 for 5-7 days at 37 ^o C, then suspended in physiological sodium chloride at a concentration 5 billion microbial cells in 1 ml. The drug is prepared by mixing equal parts of a suspension of bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT".

The antagonistic activity of the drug is determined by the disc method. To do this, the test microorganism is sown with a continuous lawn on the surface of the MPA in the cups. Sterile filter paper disks are impregnated with a suspension of the bacterial strains Bacillus subtilis TNP-3-DEPT or Bacillus subtilis TNP-5-DEPT, dried in a thermostat and applied to the surface of the agar seeded with the test culture. Crops are cultivated at 37 ^o C after 24 hours are recorded in the zone of growth inhibition around the discs. Suspension of bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT" form a zone of growth retardation in diameter of 15-25 mm of the causative agent of intestinal infections - E. coli. The bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT" show pronounced antagonistic activity against gram-positive and gram-negative bacteria. As test cultures tested certified strains: St. aureus (strain 209P), Str. equi H-34-DEPT, Sal. abortus equi BN-12, E. Coli (strain 1257), Sal. Pullorum (strain 106), Sal. typhymurium, Sal. dublin, Shigella flexneri 6783.

 To determine prophylactic effectiveness, 90 heads of newborn calves were selected during the period of mass calving of cows. The first group (45 animals) is given individually with the first portion of colostrum, 10 ml of a suspension of the bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT" 1 time per day for 3 days. The control group (45 goals) does not receive the drug. At the end of the period, there is observed (within 10-15 days) among the calves treated with the drug, signs of diseases of the gastrointestinal tract are not observed. In the control group, up to 33% of calves show signs of diarrhea.

 To determine the prophylactic efficacy of the drug in piglets, 80 sows were selected and divided into 2 groups: experimental and control. Pigs of the experimental group (40 goals) are given individually from the first hours of life 5 ml of a suspension of the bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT" 1 time per day for 3 days. The control group (40 goals) does not receive the drug. At the end of the observation period (at the time of weaning), the prophylactic effectiveness from the use of the drug was 96.1%. Moreover, in the experimental group, an increase in weight of 32.4% was noted in comparison with the control.

 During the experiment, feces are examined on days 1.3 and 10 after birth. On the first day of life in the intestinal biocenosis of clinically healthy newborn calves and piglets, lactose-positive Escherichia coli, aerobic spore-forming bacteria and enterococci predominate, microorganisms of normal microflora - bifidobacteria and lactobacilli - are poorly represented. The use of a suspension of bacterial strains "Bacillus subtilis TNP-3-DEP and TNP-5-DEP" from the first days of life helps to increase the number of the main representatives of the normal intestinal flora - bifidobacteria and lactobacilli compared with the control group.

 Thus, the use of a suspension of bacterial strains "Bacillus subtilis TNP-3-DEPT and TNP-5-DEPT" from the first days of life inhibits the development of pathogenic and conditionally pathogenic microorganisms, normalizes intestinal microbiocenosis and can be used as a method of preventing diarrhea in newborn animals.

Claims (1)

A method for preventing diarrhea of newborn calves and piglets by administering a microbial preparation inside with colostrum, characterized in that a suspension of the Bacillus subtilis TNP-3-DEPT and Bacillus subtilis TNP-5-DEPT strains in equal proportions is used as the microbial preparation.