Biotechnology of Newly Created Bacterial Composition for Siloing Based on Lactic Acid Bacteria

Abstract

Peculiarities of high-quality silage production are the use of biological products based on lactic acid bacteria. The composition of such starters varies greatly according to the use of bacterial cultures, so among the starters available on the market, the range of their effectiveness is also different. It is very common to use a one-sided approach to the choice of bacterial components, which in combination with imperfect production technology have low preservative activity. The study of combined preparations, which combine homo- and heterofermentative types of lactic acid fermentation, allows to stabilize the preservative properties throughout the ensiling time, and increase the aerobic stability of the silage after access of oxygen. Aim. Development of biotechnology of bacterial preparation for corn ensiling, optimization of cultivation conditions of newly created bacterial composition, and selection of cryoprotectants for its lyophilization. Methods. The combined preparation was created on the basis of heterofermentative strain Lactobacillus buchneri 3806 combining it in two- and three-strain compositions with other representatives of lactic acid bacteria, which are characterized by obligate homofermentative and facultative heterofermentative types of metabolism. Optimization of the environment and technological parameters was carried out using a central-compositional plan, further statistical analysis of the obtained data and determination of optimal values of input parameters according to the created mathematical model of optical density response. The effectiveness of the selected protective media was tested for the survival of bacteria after lyophilization. Results. The most effective bacterial composition was found during experiments: L. buchneri 3806, Enterococcus faecium C-8-12, L. plantarum 3216. The effectiveness of the obtained composition was tested by laboratory silage of corn. Tests of the drug based on the selected bacterial composition showed an improvement in the chemical composition of the silage compared to the untreated control and treated only with monoculture L. buchneri 3806, namely: there was a decrease in dry matter loss by 2.21% and 2.04%, 22 due to the increase of lactic acid content, and increase of aerobic stability of silage – 341 h against 57 h of the control sample, and 313 h in case of using monoculture. For the obtained bacterial composition, the culture medium of the following composition was optimized: base (hydrolyzed milk with the addition of the following components: monosubstituted potassium phosphate – 2 g/L; 5-aqueous manganese sulfate – 0.05 g/L; 7-aqueous magnesium sulfate – 0.2 g/L; twin-80 – 1.0 g/L); glucose – 19.7 g/L; yeast extract – 7.8 g/L; corn extract – 23.6 g/L; peptone – 9.1 g/L; sodium citrate – 6.6 g/L; sodium acetate – 3,4 g/L. Cultivation of the bacterial composition on an optimized medium made it possible to obtain the maximum biomass yield, at which the optical density was 2.01 units, which is almost twice as much as the value obtained by culturing the same composition in MRS medium. The optimal technological parameters of culturing the bacterial composition were established, namely the best growth was observed at a temperature of 36.4±0.4°C with constant maintenance of the pH value in the culture medium at the level of 6.5±0.1 units. In addition, the optimal composition of the protective medium containing sodium citrate, sucrose and agar was selected, and ensures the survival rate of lactic acid bacteria 98.4% after lyophilization. Conclusions. The newly formed bacterial composition can be used for the production of preparations for corn silage, and tested on other raw materials, in particular on some perennial legumes (alfalfa, clover), and the conditions of its production can be used to scale the technology.