To maintain a stable composition of the gastrointestinal tract microflora in farm animals it is necessary to use probiotic agents to ensure the full functioning of the digestive, hormonal, and immune systems of the body. Most modern probiotics include lactic acid bacteria and bifidobacteria, which are the most physiologically valuable components of a healthy organism’s an indigenous microflora. The aim of this study was to provide indication and identification from the milk of healthy cows and gastric tract of healthy pigs and calves of the genus bacteria Lactobacillus, Bifidobacterium, and Lactococcus. The objects of research were cultures of microorganisms isolated from cows milk (82), the gastrointestinal tract of cattle (317), and piglets of different age groups (114). Bacteriological studies were carried out on the basis of the veterinary sanitation and parasitology laboratory of the National Scientific Center "Institute of Experimental and Clinical Veterinary Medicine" (Kharkiv) in accordance with current regulatory documents. According to the research of the gastrointestinal tract of clinically healthy calves and piglets isolated and typified to 317 and 114 cultures of microorganisms, the species composition of the microflora (82 bacterial cultures) of the cisternous and parenchymatous milk of clinically healthy cows was determined. A total of 513 isolates of microorganisms were isolated, including: Enterobacter spp. –2 (0,39%), Staphylococcus spp. – 7 (1,37%), Bacillus spp. – 11 (2,14%), Enterococcus spp. – 33 (6.43%), Lactococcus spp. – 75(14,62%), Bifidоbacterium spp. – 170 (33,14%), and Lactobacillus spp. – 215 (41,91%). In the study of the biological properties of isolated microorganisms Lactobacillus spp. (215) established their species identity: L. brevis – 7 (3.26%), L. delbrueckii – 9 (4,19%), L. acidophilus – 21 (9,77%), L. fermentum – 23 (10,69%), L. casei – 57 (26,51%), and L. plantarum – 98 (45.58%). Cultures of Bifidobacterium spp. (170) belong to B. suis – 2 (1,18%), B. breve – 7 (4,12%), B. lactentis – 15 (8,82%), B. bifidum – 21 (12,35%), B. longum – 22 (12,94%), B. infantis – 25 (14,71%), and B. adolescentis – 78 (45,88%). From samples of biological material of farm animals, 75 cultures of the genus Lactococcus spp. were isolated (75) of which Lactococcus lactis is representative. Isolated bacteria Lactobacillus spp., Bifidobacterium spp. and Lactococcus spp. promising when creating innovative probiotic products for farm animals.
The aim of this work was to develop a technological regulation for the production of probiotic based on lacto- and bifidobacteria in the form of intestinal-soluble capsules for farm animals. Three experimental series of probiotic drug were made in the form of enteric capsules based on lactic acid bacteria. It has been established that the Bifidobacterium adolectentis No 17-316 strain and the Lactobacillus plantarum No 7-317 strain are the most adapted for the bacterial consortium. These strains have high biological activity and correspond to certain authentic characteristics. The pharmaceutical composition is proposed in the form of capsules with following ingredients: 50% of dry biomass of Lactobacillus plantarum No 7-317 and Bifidobacterium adolectentis No 17-316 strains, 5% of glucose, 5% of lactose, 9% of aerosil and 31% of starch. The content of lyophilized bacteria in one capsule is not less than 1 × 107 CFU of bifidobacteria/cm3 and 1 × 107 CFU of lactobacilli/cm3. A scheme of the manufacturing process of the drug in form of capsules includes the following stages: production of nutrient media and working solutions; growing cultures of lacto- and bifidobacteria, freeze drying of cultures of lacto- and bifidobacteria, obtaining a dry mass of lactic acid bacteria for encapsulation, encapsulation of the drug, control of the manufactured product before release, marking, packaging of capsules, transportation, storage of a drug.
Despite the success achieved in the comprehensive study of probiotic cultures, today there are a number of problems associated with the low viability of lactic acid bacteria during their processing and long-term storage in probiotics. Our work aimed to select the optimal technological regime and cryoprotectant to preserve the viability of lactic acid bacteria Lactobacillus spp. during their lyophilization. According to the results of the conducted researches, it is established that for freeze-drying of probiotic cultures Lactobacillus spp. in the facility LZ-45.27 (Frigera, Czech Republic) the most optimal is the mode which provides a rise of temperature within 45 hours from minus 70.0±1.0 °C to plus 26.0±1.0 °C with a speed of 2.2±0.1 °C/hour. It is effective to use protective media for lactobacilli, which consist of: skim milk (90%) and sucrose (10%); skim milk (90%) and lactose (10%); skim milk (90%), glucose (2.5%), sucrose (2.5%), lactose (5.0%) (P≤0.05). Freeze-drying of lactic acid bacteria under optimal conditions and the addition of cryoprotectants will avoid the problems associated with a significant reduction in the number of microbial cells. The results of research can be used for long-term storage of cultures of lactobacilli by their lyophilization.
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