The Journal of V. N. Karazin Kharkiv National University, Series "Biology"

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ISSN / EISSN : 2075-5457 / 2220-9697
Current Publisher: V. N. Karazin Kharkiv National University (10.26565)
Total articles ≅ 75
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O. A. Avksentieva, O. I. Vinnikova , V. V. Zhmurko
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 120-130; doi:10.26565/2075-5457-2020-34-13

The paper presents the results of the study of the ability of introduced Escherichia coli strains to colonize different niches of the phytosphere – the rhizosphere, endosphere, and phylosphere of Mersia winter soft wheat seedlings and to influence the plant growth response under these conditions. The ATCC 8739 and clinical E. coli strains were used as the study material, as well as an isolate from the wheat agrocenosis soil. The vegetation experiments were carried out in the factorial chamber of the Department of Plant Physiology and Biochemistry of Plants and Microorganisms of V.N.Karazin Kharkiv National University. The experimental plants were inoculated with suspensions of E. coli strains and soil isolate by watering them in vegetation vessels where seedlings were grown for 10 days. In parallel by the method of successive washes, we analyzed the dynamics of the number of E. coli CFU in the rhizome, endo- and phyllosphere of seedlings. After completion of the experiment, the growth response was analyzed by linear growth and by the integral index of growth and biosynthetic processes – biomass accumulation. The results of experiments show that inoculation with ATCC 8739 and clinical strains of E. coli reduced the germination of wheat seeds and inhibited the growth response. Inoculation with E. coli soil isolate has virtually no effect on seed germination, linear growth, and biomass accumulation by Mersia seedlings. It was found that the number of E. coli bacteria in the phytosphere of wheat plants depended on the type of inoculated strain and differs in different parts of the seedlings phytosphere. The maximum number of E. coli CFU in the phytosphere was detected at the inoculation of plants with the clinical strain, three times less – at exposure to the soil isolate, almost 6 times less – at the inoculation with the standard strain. Differences in the degree of colonization of different parts of the phytosphere were detected at the inoculation of the test seedlings with different strains and soil isolates of E. coli: in the control variant and under the influence of ATCC 8739, the distribution between the rhizo- and endosphere is approximately the same. When the plants were inoculated with the clinical strain, endosphere was colonized, when using the soil isolate – the rhizosphere. In the phyllosphere of the experimental seedlings, only cells of the clinical strain were found in a small number, which indicates its increased adhesive properties. Plant-microbial relationships and the ability of conditionally pathogenic E. coli bacteria to colonize different areas of the plant organism and to use wheat seedlings as an alternative host are discussed.
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 112-119; doi:10.26565/2075-5457-2020-34-12

Ultrastructural features of the nematode Trichostrongylus tenuis were studied. Helminthological studies were carried out in the Nakhchivan Autonomous Republic in 2015–2018. The material from domestic waterfowl was collected by the method of complete parasitological dissection. Along with the fact that this helminth is a specific parasite of domestic waterfowl, it dominates among all the types of helminths noted by us and is the cause of serious changes in the host organism. The study of the ultrastructure of the nematode T. tenuis is important for identifying parasitic-host relationships, clarifying the systematic position of parasites and in preparing measures to combat these parasites. In addition, a wide variety of morphological features of the lateral alae, allows them to be used as one of the main features for identifying helminth species. The article first presents data on the ultrastructural features of the lateral alae of the nematode T. tenuis. As a result of the studies, it was found that, despite the fact that in some parasitic nematodes of the Trichostrongylidae family the morphological structure of the lateral alae of the cuticle is identical over the entire surface of the body, in the nematode T. tenuis belonging to the same family, four forms were revealed during ultrastructural studies, which were represented using diagrams and electron diffraction patterns. Lateral alae consist of cortical, homogeneous, and fibrillary layers differing in size, thickness, and other features. From the front (oral) part of body of the nematode T. tenuis to the beginning of the intestine, the cuticle is smooth; on the body part from the small intestine and behind it, lateral alae begin to be observed, which resemble a “ridge” in shape. When approaching the posterior end of the body, the lateral alae take the form of “hooks”. On the caudal region, the lateral alae become even more complicated and get the form of “spikes”, and relatively small outgrowths are additionally observed on these lateral wings. These signs can be used to clarify the taxonomic position of helminth species.
S. M. Okhrimenko , A. Yu. Grishkova
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 28-34; doi:10.26565/2075-5457-2020-34-3

The accumulation of heme in the organism under the influence of various hemolytic factors can cause the development of oxidative stress with the activation of free radical processes, oxidative damage to macromolecules and supramolecular complexes of cells and tissues. Under these conditions, the antioxidant defense system is activated in the organism, an important link of which is thiol compounds, particularly glutathione. Under such conditions, the processes of nitrogen and carbohydrate metabolism associated with the formation of adaptive reactions in response to stress have been investigated insufficiently. The aim of this work is to study some indicators of nitrogen and carbohydrate metabolism during the administration of hemin and the combined administration of hemin and glutathione to clarify the role of this antioxidant in the possible correction of metabolic processes. The subjects of the study were mature outbred albino male rats that received intraperitoneal injections of hemin (50 mg/kg) and glutathione (500 mg/kg) solutions, which was administered 0.5 hours before the introduction of hemin. The animals were tested 2 hours after hemin administration. The content of total and non-protein -SH groups, and the activity of gamma-glutamyltranspeptidase (GGT) in liver and kidney homogenates, glycogen content and tyrosinaminotransferase (TAT) activity in liver homogenate were studied. The content of reduced -SH groups can be an indicator of pro-antioxidant balance, GGT activity is one of the indicators of glutathione metabolism, and glycogen content and TAT activity in liver are hormone-sensitive indicators. The introduction of hemin caused a decrease in the content of total and non-protein -SH groups, glycogen content and an increase in TAT activity in liver, as well as an increase in the activity of GGT in this organ. Administration of glutathione to rats 30 minutes before the administration of hemin prevented shifts in these parameters in liver caused by the administration of hemin alone. In kidneys, an increase in the content of total -SH groups was found after the combined administration of glutathione and hemin compared with the effect of hemin alone. The results of this study may indicate a sensitivity of nitrogen and carbohydrate metabolism in rat organs to the effect of hemin and the corrective effect of glutathione under these conditions, probably mediated through an increase in the thiol component of the antioxidant defense system.
N. B. Hryntsova, A. M. Romanyuk, Mykola Serhiiovych Lyndin , T. V. Ryabenko
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 141-150; doi:10.26565/2075-5457-2020-34-15

Xenobiotics, including salts of heavy metals, determine the development of pathology of individual organs and systems of the body. Adrenal hormones occupy a key place in the regulation and maintenance of the organism basic functions. The study of morphological, biochemical and immunohistochemical changes in the adrenal cortex of pubescent male rats at exposure to the complex of heavy metal salts remains an actual aspect of modern morphology. The experiment involved 24 white adult male rats weighing 250–300 g, aged 7–8 months. The animals of the experimental group consumed ordinary drinking water saturated with a combination of heavy metal salts (Zn, Cu, Fe, Mg, Cr) for 60 days. Histological, biochemical (determination of the content of hormones COR – cortisol, DHS – dehydroepiandrosterone sulfate) and immunohistochemical (determination of the expression of proliferation marker Ki-67) methods were used. Long-term consumption of the combination of heavy metals salts by mature rats leads to thickening of the stromal component of the gland, the rupture of capillaries and the violation of rheological properties of blood. Chromatin condensation, its marginal location, and initial stages of necrobiotic changes have been found in the spongiocyte nuclear apparatus. For endocrinocytes of the zona glomerulosa and zona reticularis of the gland, a slight increase in proliferative activity has been revealed, while cells of the zona fasciculata were non-reactive. These results correlate with the results of biochemical research, according to which hormones of the zona reticularis – DHS were actively involved in the formation of adaptive reactions in the body of experimental animals at the 60-day period of administration of the complex of heavy metals salts. The results of complex studies indicate a weakening of the secretory activity of cells of the zona fasciculata, which adversely affects the development of compensatory-adaptive processes in the body and the course of the general adaptation syndrome in response to the action of the damaging agent.
I. V. Nikitchenko , T. P. Rybalchenko, T. V. Barannik , O. V. Pavychenko
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 18-27; doi:10.26565/2075-5457-2020-34-2

Oxidative stress is considered to be the main mechanism of cadmium ions toxic effect on the cells and is caused by cadmium, as a non-transition metal, indirectly. Oxidative damage to cells due to the action of cadmium ions is tissue-specific and is associated with the antioxidant system inhibition, free heme accumulation and essential metals substitution in metalloproteins. Nitrogen monoxide (NO) exhibits high affinity for heme and proteins and peptides sulfhydryl groups, known to be the main molecular targets for cadmium ions. Taking all the above-mentioned into account, the aim of this work was to study the effect of NO radicals donors on the prooxidant-antioxidant state of mammalian tissues under oxidative stress caused by cadmium chloride administration in vivo. Male Wistar rats weighing 160–200 g were used in the study. CdCl2 was administered subcutaneously at a dose of 14 mg/kg body weight. The direct donor of the NO radical sodium nitroprusside (SNP, 1 mg/kg mass) and the substrate of the NO synthase reaction L-arginine (600 mg/kg mass) were administered intraperitoneally. In order to study the corrective action, donors of the NO radical were injected 0.5 h before the cadmium salt. The objects of investigation were blood plasma and liver, kidneys and spleen homogenates of rats. The cadmium chloride treatment caused a number of prooxidant-antioxidant balance disorders, most of which were revealed a day after injection. The accumulation of lipid peroxidation products was found in rat serum, liver, and spleen. The enhancement of prooxidant processes in these tissues may originate from cadmium ions and hemolysis products entry. In the antioxidant system, significant changes were observed under cadmium action only in the liver: an increase in the reduced glutathione content and SOD activity and a decrease in catalase activity. The precursor of nitric oxide L-arginine did not change the basal level of prooxidant-antioxidant parameters, and in most cases did not affect their dynamics in the organs studied after cadmium chloride administration. A direct NO donor, sodium nitroprusside, acted in liver and spleen mostly as a prooxidant. In liver, the injection of only nitroprusside, as well as the combined administration of SNP and CdCl2, led to free radical processes activation just in two hours. In spleen, the combined treatment by SNP and cadmium salt also caused an earlier development of oxidative stress, as witnessed by an increase in lipid hydroperoxides level and a decrease in reduced glutathione content. Therefore, the injection of a direct NO donor, sodium nitroprusside, and a substrate of NO synthase, L-arginine, in selected doses has insignificant corrective action on cadmium-induced oxidative stress in the liver, kidneys and spleen. However, in blood both donors of NO effectively prevented the accumulation of lipid peroxidation products under CdCl2 treatment; in addition, L-arginine significantly reduced the lactate dehydrogenase release, which may indicate blood cells and blood vessels protection from the damage caused by cadmium ions.
P. P. Pukhtaievych, K. P. Kukol , N. A. Vorobey , S. Ya. Kots
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 163-174; doi:10.26565/2075-5457-2020-34-17

Under the conditions of a model pot experiments, the reaction of the self-fertile lines of alfalfa Kishvardy 46, Kishvardy 27, Vertus and Ziguen to inoculation with nodule bacteria Sinorhizobium meliloti AC48 and AC88 was studied. As a result of studies, it was found that the intensity of assimilation of N2 by symbiotic systems created with the participation of various genotypes of alfalfa and active strains of S. meliloti is one of the main factors that affects the vegetative mass yield of this important forage crop. Self-fertile lines of Medicago sativa L. plants, inoculated with different strains of rhizobia were characterized by higher rates of the mass formed on the root nodules, compared to the control plants of the alfalfa variety Yaroslavna. The traditional dynamics of nitrogen-fixation activity of root nodules was maintained in all the symbiotic systems studied by us, with low values in the stems formation stage and intensive growth in the budding and flowering stages. The highest level of nitrogen fixation and vegetative growth of plants (values of plants green and dry mass, roots and root nodules mass) was established by inoculation of alfalfa line Kishvardy 46 with strain S. meliloti AC48. During the growing season the indices of the mass of nodules formed on the roots of these plants were higher by 1.8–2.3 times, the green mass by 1.2–1.6 times and the height of the plants 1.2–1.4 times as compared to the control. In the flowering stages the nitrogen-fixation activity of the symbiotic complex of plants of the Kishvardy line 27 and nodule bacteria S. meliloti AC48 exceeded the values in the symbiotic systems formed with the participation of the same strain and plants of the Ziguen and Vertus lines by 13.0 and 39.4 %. The lowest values of nitrogen fixation activity were observed by inoculation of plants of the Vertus and Ziguen lines with active strains S. meliloti AC48 and AC88 compared to the symbioses formed by the plants of the Kishvardy lines 27 and 46, as well as of the control-variety Yaroslavna with the noted strains. A stimulating effect of inoculation of alfalfa seeds of different genotypes on the growth and development of plants was noted, as evidenced by the positive dynamics of the increase in above-ground mass, the accumulation of dry matter and higher than the control values (indicators) of plant height during the growing season.
S. A. Katrushenko
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 78-88; doi:10.26565/2075-5457-2020-34-9

In Ukraine, amphibian anomalies have not been sufficiently studied. The most studied regions are Kyiv region and the steppe zone of Ukraine. In the Kharkiv region, studies of external anomalies of amphibians have not been conducted before. In 2016 we have begun to study this problem. The purpose of this study is to examine the variety of external anomalies of amphibians in the Kharkiv region. The material is sampling amphibians from the stock collections of the Museum of Nature of V.N.Karazin Kharkiv National University and sampling live amphibians from different localities of the Kharkiv region, explored in the period from 2016 to 2018. Anomalies were determined visually, according to the classification of V.L.Vershinin and O.D.Nekrasova. For each the amphibian species frequency of malformed animals Pas (the proportion of abnormal individuals from all individuals in the sample) and partial frequency of abnormalities Ap (the proportion of individuals in the sample having this anomaly) were determined. As a result 17 variants of morphological abnormalities were identified: brachydactyly, oligodactyly, ectromelia, polyphalangy, finger thickening, rotation, syndactyly, shizodactyly, ectrodactyly, skin web and growth on the foot, thaumelia, hemimelia, lack of a swimming membrane, anophthalmia, brachycephaly and pattern abnormal. The most widespread anomaly encountered in the surveyed amphibians is brachydactyly (accounting for almost half of the reported cases of morphological abnormalities). This limb anomaly is the least harmful, so individuals with such abnormalities are usually quite viable. Amphibian anomalies are evenly distributed among samples: the highest occurrence of individuals with anomalies was recorded in Lissotriton vulgaris, in other samplings of frog (except Bufotes viridis and Bombina bombina), the occurrence of individuals with anomalies did not exceed five percent. A new type of body color anomaly in this year's Bufotes viridis was also identified and described.
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 35-42; doi:10.26565/2075-5457-2020-34-4

Earlier we have shown that dirhenium(III) dicarboxylate complex with γ-aminobutyric acid possessed higher antitumor activity, than those of the previously investigated alkylcarboxylates, also may act as a modulator of cisplatin mechanism of action and as a stabilizer of red blood cells in tumor-bearing organisms. Thus, the task of the work was to investigate anticancer activity of the complex cis-[Re2(β-Ala)2Cl6] (I) in the model of tumor growth in vivo and to realize if the amino acid residue influences the DNA-binding activity of the amino acid derivatives of the cluster rhenium(III) compounds. Antitumor properties of the complex I were studied in the model of tumor growth with the use of Wistar rats inoculated by tumor carcinoma Guerink cells. The introduction of the compound alone in free and liposomal forms inhibited the tumor growth by 36 % and 45 % correspondingly, that is more than for dirhenium(III) clusters with alkyl ligands. The combined introduction of I and cisplatin had a significant impact on the tumor growth and showed the disappearance of the tumors in most of the animals. No considerable differences were found between introduction of liposomal and free form of I. The electronic absorption spectra of Calf Thymus DNA (CT-DNA) exhibit hyperchromism in the presence of increasing amounts of I. The DNA band at ~ 260 nm arises from the π-π* transitions of the nucleic acid bases and changes in the intensity and slight wavelength shifts of this characteristic band reflect the corresponding structural modifications of the DNA, which include changes in stacking, disruption of the hydrogen bonds between complementary strands, covalent binding of the DNA bases, intercalation of aromatic rings and others. The binding constant Kb(I) = 2.43 × 103 M-1 to CT-DNA was obtained that was lower than the values reported for the classical DNA intercalators and compares well with the magnitude of the binding constants for other complexes of dirhenium(III); titration of СT-DNA with cisPt and hydrogen peroxide also leads to a hypochromic effect, weak at low concentrations and more significant at high concentrations of I; the DNA binding constants increased in several times when using H2O2 or cisplatin that confirms a mechanism for redox activation of interaction of I with DNA in a cancer cell. The obtained results demonstrate the possibility of application of the amino acid derivatives of dirhenium(III) clusters in antitumor therapy.
M. O. Baranets, I. I. Korshуkov
The Journal of V. N. Karazin Kharkiv National University, Series "Biology", Volume 34, pp 43-51; doi:10.26565/2075-5457-2020-34-5

We ascertained the features of formation of self-renewing and self-sustaining of the local population of the Mediterranean introducent Hyssopus officinalis L. in the extremal ecologic and edaphic conditions of an iron ore dump in Kryvyi Rih Area, seeds of which have been sown on the area of 1 ha about 30 years ago. It was found that the species, during this period after reaching the reproductive phase of development, has taken the area of the dump of about 3 ha and, in fact, it formed the isolated population. This species annually multiplied by seed, and spread on the areas of ore dump, where almost no vegetation cover, in fact, performing a pioneer function. At the site, where the initial sowing of H. officinalis seeds was carried out, the phytocoenosis forms now; this phytocoenosis, according to its floristic composition, approaches the natural steppe, but the introduced species are gradually ousted. In habitats occupied by the species due to its spontaneous distribution, successional processes are at an initial stage. Within some of the population loci of H. officinalis, we have found up to 513 adult individuals with the number of generative sprouts from 20 to 70 per a plant. Flowers are formed on these sprouts in average from 160.2 to 181.6. Average seed productivity is 60 % of theoretically possible rate, and the seeds from the parent species extend to a distance of 1–5 m. In a compact areas where H. officinalis grows, there are plants of all the age groups – from the seedlings to the subsenile. H. officinalis has naturalized in the dump conditions, and its local population is а resistant, full-grouped; it develops under the conditions where other species are not competitive. Generatively developed plants are higher than 90 cm and have good vital state. The species on the dump shows pioneering features; it is not aggressive and can be displaced by zonal vegetation; thus, it does not pose invasion risk. H. officinalis can be recommended for the revegetation of iron ore dumps, or their large decommissioned areas, without technical preconditioning, only by direct sowing of the seeds into the dump rocks, to promote its spread artificially.
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