Effect of metal microelements on biochemical indices of probiotic bacteria

The influence of metal-microelements (Zn, Cu, Ag, Au) in colloid and ionic form on the main biochemical parameters (ATP-ase activity, transmembrane potential and respiratory activity) of E. coli G35 N#1-413 and Ent. faecalis G35 N#4-410 probiont-strains has been studied with the goal to create complex metal-bearing probiotic preparations. Monotonous dose-dependent inhibitory influence of all mentioned metals in ionic form on the bacteria functional activity has been established. Metals' colloids had certainly stimulating influence on microorganisms' biochemical parameters in the same concentration limits with the following maximum positive effects: for Zn - 2x10(-7)-2x10(-4) mg/ml, for Cu - 8.4x10(-7)-8.4x10(-6) microg/ml, for Ag - 23x10(-7)-23x10(-4) microg/ml, for Au - 5x10(-6) microg/ml by metals. It has been shown, that cultural characteristics of probiont-bacteria in the presence of both ionic and colloid forms of studied metals remain stable. Certain stimulation of probiont-bacteria' main enzymes and life-support processes by colloidal forms of the studied metals-microelements in determined concentration and particle dimension limits is the evidence of a possibility of metals' colloids application in probiotic preparations' composition in order to increase the resistance and functional activity of probiont-microorganisms.     

Effect of ionic and colloid gold on ATP-hydrolase fermentative systems in the membrane of Bacillus sp. B4253 and Bacillus sp. B4851

Accumulation of gold in cells of Bacillus sp. B4253 can be directly or indirectly connected with activity of bacteria plasma membrane basal Mg2+-ATPase. Therefore this work deals with a comparative analysis of kinetic properties of plasma membrane basal azide-resistant Mg2+-dependent ATP-hydrolase activity of B. sp. B4253 and B. sp. B4851 capable to gold accumulation and not capable to this process, accordingly. It is shown, that by a number of kinetic parameters - specific fermentative activity, initial speed of reaction of hydrolysis ATP (V0), Mixaelis constant (Km), the maximal initial speed by Mg2+ (V(Mg)) and by ATP (V(ATP)), optimum concentration of ATP ([ATP]opt), pHmax, sensitivity to action of the thapsigargine and eosine Y - bacteria membranes basal Mg2+-ATPase activity accumulating gold, and the bacteria not capable to this process, are identical. But by some parameters they differ: Mg2+-ATPase activity of membranes of the bacteria which do not accumulate gold, has three times greater affinity for Mg ions and smaller value [Mg]opt. The inhibition effect of ionic gold (10(-4)-3x10(-4) M) is shown on azide-sensitive (H+-ATPase) and azide-resistant (Mg2+-ATPase) components Mg2+-dependent ATP-hydrolase activity in fraction of plasma membranes of microorganisms Bacillus accumulating gold, and not capable to this process. Colloid gold (0.0002-4 microg/ml) stimulates activity of H+-ATPase and Mg2+-ATPase in a membrane of the bacteria accumulating gold 1.5-2 times, and does not influence activity of ATPases of a membrane of the bacteria which do. not accumulate gold.     

Quantification of phenol compounds using Pseudomonas free bacterial cells

The possibility of using Pseudomonas free bacterial cells for integral evaluation of some phenol compounds' content in water by means of estimation of specific respiratory activity (SRA) value was shown. Amperometric system of Clark electrode was used as a transducer in conducted investigations. The conditions of performing the SRA analysis of Pseudomonas sp. B 4251 and Ps. putida sp. microbial cells were optimized. The method sensitivity is 0.2 mM for Pseudomonas sp. B 4251 and 0.5 mM for Ps. putida sp. Taking into account specificity and sensitivity of the studied method, for estimating the integral toxicity of waste water, which contains 4-nitrophenol and 2,4-dinitrophenol in more than MPC concentrations and harmful for ecosystem became possible.     

Identification and catalytic properties of Mg2+-dependent ATP-hydrolase of cytoplasmic membrane of Bacillus sp. B4253 capable of gold accumulation

Ca2+-independent, Mg2+-dependent ATP-hydrolase fermentative activity consisting of two components--azide-sensitive and azide-resistant ones has been identified in cytoplasmic membrane of Bacillus sp. B4253 capable to gold accumulation in ionic and colloid forms. The authors have characterized properties of the azide-resistant component of ATP-hydrolase reaction: dynamics of accumulation of one of the reaction products--inorganic phosphate P(i); dependence of ATP hydrolysis rate on the membrane protein content; pH-dependence; sensitivity of ATP-hydrolase activity to the change of reagents (ATP, Mg2+) concentration, as well as to the effect of some specific and nonspecific inhibitors of ion-transporting Mg2+-dependent ATP-hydrolase systems (ouabain, tapsigargin, eocine Y, La ions). It is supposed that the obtained experimental data can be used for the following study of molecular and membrane mechanisms of gold accumulation in Bacillus sp. B4253.     

Veterinary probiotic preparation influence on the biochemical blood indices of young farm animals

The veterinary probiotic preparation was developed on the basis of aerobic part of bacterial strains of normal human intestinal microflora: 3 strains of Escherichia coli--G35No1-413, G35No2-412, G35No3-411, and one Enterococcus faecalis G35No4-410 strain. The influence of a pilot lot of the prepapration on the biochemical blood indices and the state of intestinal microflora of different age groups of young farm animal (calves and piglets) was studied during the process of the commission tests. It has been shown, that the developed probiotic promotes the improvement of protein and mineral metabolism, normalization of level of enzyme activities, which show state of the native immunity of the young farm animals' organism. It was the result of normalization of the intestinal microflora composition. The optimal doses and periods of intake have been determined. On the basis of executed researches the veterinary probiotic preparation has been presented for registration to the State Department of Veterinary Medicine of Ukraine.