Mitochondria are organelles that are most sensitive to the action of stressors on any cell of the entire organism and exposure to chemicals which can cause its dysfunction and cell death in general. Especially sensitive to adverse conditions are liver mitochondria, where the processes of biotransformation of endogenous and exogenous metabolites are formed, not only in the liver, but also in other organs and tissues. Mitochondrial dysfunction can cause instant hepatic cytolysis and steatosis. Therefore, early detection of mitochondrial toxicity is important during preclinical studies of new pharmacological agents, as this will help avoid remote negative effects. The biologically active feed additive Humilid, a complex of humic acids known for their antidiarrheal, analgesic, immune-stimulating, and antimicrobial properties; shows a corrective effect on the activity of the lysosomal cathepsin; enhances the positive effect of hematopoiesis on hemoglobin and its quality indicators consisting of red blood cells; and activates the synthesis and accumulation of fibronectin expression that takes part in the formation of immunological protection of animals. The objective of our experiment was to determine the effect of complex biologically active feed additives based on humic substances on the biochemical indicators of the liver mitochondrial antioxidant system of Mongolian gerbils (Meriones unguiculatus Milne-Edwards, 1867). The experiment was conducted on mature (6 months) Mongolian gerbils. The data obtained showing the influence of the biologically active feed additives Humilid, alone or in combination with ascorbate and Eco-impulse Animal, on the antioxidant defense system of liver mitochondria of gerbils are presented in this article. The proven antioxidant effect of humic substances in the mitochondrial fraction of the liver which inhibits the accumulation of oxidized products in the cells is shown, confirmed by the decrease in the number of TBA-active products, catalase activation, and an increase in the concentration of cytochrome C. Also, an increase in the amount of cytochrome C, which is a direct participant in the mitochondrial respiratory chain and provides efficient electron transport, indicates the acceleration of energy supply processes. The functional activity of mitochondria was accompanied by increased activity of aspartate aminotransferase involved in the shuttle of malateaspartate transport of electrons through the mitochondrial membrane. The results obtained indicate the positive reaction of gerbil liver mitochondria under the influence of the biologically active feed additives of humic substance.Keywords: Humilid; ascorbic acid; Eco-impulse Animal; aspartate aminotransferase; catalase; superoxide dismutase; TBA-active products; cytochrome C Вплив біологічно активних кормових добавок гумінової природи на систему антиоксидантного захисту мітохондрій печінки монгольської піщанки О. О. Дьомшина*, Г. О. Ушакова*, Л. М. Степченко** *Дніпровський національний університет імені Олеся Гончара, Дніпро, Украї...

Doxorubicin causes toxic side effects which result in profound changes in various organs including the heart muscle, which leads to the development of cardiomyopathy, with further complications, as our earlier studies have shown. But there are complications in treatment by anthracycline chemotherapy. The reduction of toxicity of cytotoxic drugs without reduction of their antitumor action remains an important medical challenge. The S-100b and glial fibrillar acid protein (GFAP) are the key astroglial proteins which are widely used as biomarkers for neurotraumas and strokes. Thus the purpose of our work was to study the GFAP and S-100b levels under the experimental model of doxorubicin-induced cardiomyopathy and the effect of the cytostatic separately or combined with Humilid. The level of studied proteins was measured by ELISA using monospecific antibodies against GFAP and S-100b (Sigma, USA), secondary anti-IgG labeled with horseradish peroxidase (Sigma, USA) and GFAP and S-100b standards (Sigma, USA). The experiment was conducted on white Wistar male rats weighing 210 ±50 g, which were divided into 3 groups with 8 animals in each: 1 – control, rats received saline i/p, 2 – doxorubicin 1 mg/kg i/p once weekly during 4 weeks, 3 – doxorubicin by the same scheme plus 0.01% solution of Humilid during 4 weeks. Under impact of doxorubicin an increase of fibrillar GFAP content in the rats’ hippocampus was shown, which may have occurred as a result of the proliferation of astrocytes. An increase of concentration of S-100b to 20–26% compared with the control animals was also detected in the hippocampus, thalamus and the cerebral cortex of the studied animals. Concomitant administration of doxorubicin with Humilid partially prevented the increase in concentration of GFAP and S-100b in the rats’ hippocampus. The reciprocal change in the level of S-100b and GFAP in the peripheral nervous system of the heart tissue was defined with a high degree of correlation under impact of doxorubicin. Accordingly, we have proposed the hypothesis that the increase in of the filament GFAP in the hippocampus may occur due to the development of the ischemic state under the impact of doxorubicin-induced cardiomyopathy. Humilid with doxorubicin prevented changes in the researched astroglial proteins of the heart muscle and brain parts of rats.

Humic acids are characterized by chelating properties due to which they are able to influence the mineral metabolism in animals. Rabbits have species-specific characteristics of mineral metabolism, which distinguishes them from most species of mammals. For the experiment, 16 rabbits of 45-day-old of Hyplus breed were divided into the experimental and control group of 8 rabbits in each. The animals were 15 days in the preparatory period. The rabbits of the experimental group received humic acid additive individually with water in the amount of 5 mg/kg, the rabbits of the control group received pure water in the same amount. The ability of humic acid additive to increase the content of calcium, ionized calcium, iron, to correct the content of copper and zinc, and also to increase the activity of alkaline phosphatase in the blood serum of rabbits of the experimental group has been proved. We observed the early effect of the humic acid additive on increasing the content of iron and calcium ionized and a later effect on the activity of alkaline phosphatase and an increase in the content of copper in the blood serum. The influence of humic acid additive on the distribution of calcium, phosphorus, manganese, copper and zinc in bone tissue is determined. According to a histological study, there was a significant increase in the number of layers of osteons and osteoblasts in the bone tissue of the femur, an increase in the number of osteons and osteoblasts in the bone tissue of the sternum and an increase in the number of columns of chondrocytes and an increase in the number of chondrocytes in the column in the cartilaginous tissue of the sternum. The results of a histological study of bone tissue, together with an increase in structural macroelements in it and redistribution of osteotropic trace elements in the blood and bone tissue suggest the possibility of using a humic acid additive to intensify the growth and mineralization of bone tissue, which can improve the state of bone tissue of rabbits in the postnatal period ontogenesis.

The article is devoted to the research of possibility of preventive influence of antioxidants, feed additives of humic nature on the state of the erythrocytic system under the influence of combined water-immobilization stress. The animals were divided into five groups of 6 animals each: І -the group of intact animals (control); І-V -experimental groups. The animals of all experimental groups were added «Humilid» feed supplement orally (5 mg/kg of body weight per active ingredient), each animal individually with the help of a probe for 18 days, «Eco-Impulse Animal» (2.5 mg/kg of body weight), water and vitamin E (50 mg/kg of body weight). In the later period the animals of ІІ, ІІІ, ІV and V group were conditioned under stress simulation. The blood was analyzed by the number of erythrocytes, erythrocyte content, hematocrit, color index and calculated erythrocytic coefficients. It was established that in animals of group II, after the effect of combined water-immobilization stress, there is a significant decrease in the number of erythrocytes, hemoglobin content and the percentage ratio of formed blood elements is changing. The indices in animals received before the stress period of adding «Humilid», «Eco Impulse Animal» supplements and vitamin E showed no significant deviations from the control values. However, those rats which before the stress period were added «Eco-Impulse Animal» and vitamin E supplements to the diet, showed a significant increase in the average volume of red blood cells (MCV) and the average content of hemoglobin in erythrocyte (MCH). This fact points to the ability of these substances to influence the processes of blood condensation. The comparison of the influence of antioxidants, «Humilid» and «Eco-Impulse Animal» food additives with vitamin E on the hematological parameters of blood in rats indicates the presence of almost similar effect in the studied substances. Еритроцитарна система крові щурів на тлі застосування кормових добавок гумінової природи за комбінованого стресу Л. М. Дяченко, Л. М. Степченко Дніпровський державний аграрно-економічний університет, м. Дніпро, УкраїнаАнотація. Розглянуто можливість превентивного впливу антиоксидантів, кормових добавок гумінової природи на стан еритроцитарної системи за впливу комбінованого водно-іммобілізаційного стресу. Тварин розділили на п'ять груп по 6 тварин: І -група інтактні тварини (контроль); ІІ-V -дослідні групи. Тварини дослідних груп додатково отримували перорально, індивідуально за допомогою зонду протягом 18 діб воду, кормові добавки «Гумілід» (в розрахунку 5 мг/кг маси тіла за діючою речовиною), «Еко Імпульс Animal» (2,5 мг/кг маси тіла) та вітамін Е (50 мг/кг маси тіла). У тварин ІІ, ІІІ, ІV та V груп моделювали стрес. У крові визначали кількість еритроцитів, вміст еритроцитів, гематокрит, кольоровий показник і розраховували еритроцитарні коефіцієнти. Встановили, що у тварин ІІ групи, після впливу комбінованого водно-іммобілізаційного стресу відбувається достовірне зменшення кількості еритроцитів, вмісту гемоглобіну та відсоткове...

The «Humilide»-Biologically Active Feed Supplement is a 10 % solution of humic acids that, by its antioxidant, immunostimulatory and antimicrobial properties, affects feed conversion, productivity and meat quality of farm animals. It is made according to TU In 15.7-00493675 004: 2009 in the conditions of the Research Laboratory of Humic Substances named of prof. L. A. Khristeva. Most of the work on the effects of the supplement has been done on poultry of different species and pigs. The available data on the safety of the feed additive «Humilide» were analyzed and biotested on Paramecium caudatum in accordance with the requirements of the current legislation in the field of control of veterinary preparations and feed additives. A series of Humilide dilutions of 0.1-100 g/l on the culture medium at ciliates were prepared for toxicological studies. Acute toxicity data of the tested concentrations were obtained according to the recommended methods of rapid bioassay. LC 50 values were determined by probit analysis of lethality curves. In solutions of «Humilide» 1.0 and 10 g/l within 1 hour of observation, the death of the protozoa was less than 10 % of the total number. A concentration of 100 g/l causes a decrease their quantity. According to the calculation of the toxicity of «Humilide» by the method of Bliss & Prozorovsky LC 50 for Paramecium caudatum is 104 g / l. Using the graphical method for calculating LC 50 from the obtained regression line, the result was 124 g/l. Thus, «Humilide» at a concentration of less than 1 % is non-toxic to protozoa, NOEC (No Observed Effect Concentration) is 2 g/l and EC 50 (Half maximal effective concentration) is 104-124 g/l. Therefore, the results on the activity of «Humilide» in relation to the Paramecium caudatum allow to classify it as a non-toxic substance, LC 50-over 5 000 mg/kg. It is IV class acute toxicity according to SP 8.8.1.2.002-98 (Ukraine) and international criteria-5 toxicity category. When converted to the content of humic acids in the composition of the sample, this corresponds to 10.4-12.4 g/l.