Influence of soil moisture level on metabolism of non-structural carbohydrates in Quercus robur leaves

The long-term increases in average temperature and intensification of droughts which characterise the current state of the Earth’s climate system have a negative impact on forest ecosystems and can lead to a decrease in their area and deterioration of the living conditions of their components. In the conditions of the Ukrainian Steppe an important environmental, antierosion, water-protective and soil-protective role belongs to the ravine forests. The most valuable component of the ravine forests is presented by natural populations of common oak (Quercus robur L.), which are able to tolerate the arid climate typical of the steppe region. But with global warming, the endurance of this species is changing. It is believed that a significant role in plant adaptation to drought and high temperatures may belong to non-structural carbohydrates. Therefore, it is important to study changes in the concentration of these substances in the leaves of this leading species under the action of adverse hydrothermal conditions. The article analyzes the content and dynamics of soluble sugars (glucose, fructose, sucrose) and starch in the leaves of Quercus robur L. under different forest growth conditions of the ravine forest (hygromesophilic (CL2–3), mesoxerophilic (CL1) and xerophilic (CL0)). The research was conducted in the forest in the Viyskove area (steppe zone of Ukraine) in the thalweg and at different levels of slope of southern exposure. Content of glucose, fructose, sugar and starch in Quercus robur leaves was determined. It was found that when exposed to high temperatures and increasing water stress during the vegetation period in xerophilic (CL0–1) and mesoxerophilic (CL1) forest growth conditions, the concentration of both glucose and sucrose in the leaves of Q. robur increases and it becomes much higher than in conditions of more optimal water supply. At the same time, the disaccharide content increases more significantly than that of monosaccharide. The greatest amount of these sugars is observed in the driest months (July, August), when conditions for providing plants with water are the most stressful. When water stress grows the increase in concentration of glucose and sucrose is correlated with reduction of starch content. It has been found that the concentration of fructose in Q. robur leaves in droughty conditions of growing was comparable to more favourable conditions of moisture. In September, there is a decline in the content of all forms of non-structural carbohydrates in the leaves of plants of all variants compared to the previous month, especially in conditions of adverse water supply. Therefore, forest growth conditions do not affect the nature of the dynamics of soluble sugars and starch in the leaves of Q. robur, although they change their quantitative indicators. Based on the protective function of sugars under the action of stressors on plants, we can assume that in conditions of significant lack of moisture in the soil their accumulation in the leaves in areas with mesoxerophilic and xerophilic hygrotopes plays an important role in increasing Q. robur drought resistance.

Influence of heavy metals on physiological and biochemical parameters of Pseudorasbora parva (Cypriniformes, Cyprinidae)

Anthropogenic load on aquatic ecosystems leads to increased inputs of heavy metals, which can have a toxic effect on aquatic organisms. Some of the most appropriate objects for research are short-cycle fish species. This article considers the results of studies on the adaptive reactions of the stone moroko Pseudorasbora parva (Temminck et Schlegel, 1846) to Mn, Pb, Ni heavy metal ions, which exceeded the reference values in the reservoir by 1.7, 1.5 and 2.0 times, respectively. Changes in morphological parameters of the blood and histocytological pattern of the hepatopancreas of the experimental species under the influence of toxicants, as well as changes in biochemical parameters, were determined. It was noted that the influence of Mn caused pathological changes in the form of poikilocytosis. The morphometric parameters of erythrocytes (the cell area and the nuclear area) reached 67.48 ± 0.67 and 13.97 ± 0.22 µm2 respectively (4.0% and 13.8% less compared to the control). The influence of Ni resulted in an increased number of leukocytes and immature forms of erythrocytes. The area of erythrocytes was 0.9% smaller than that of the control group, and the area of the nucleus was 4.5% smaller than in the control. The effect of Pb as well as Mn consisted in poikilocytosis. The area of red blood cells and nuclei was smaller by 6.5% and 8.3%, respectively, compared to the control. The percentage of white blood cells in fish exposed to Ni and Pb tended to increase. In the experiment with nickel, the percentage of white blood cells was 10.2% of the number of formed elements; in the experiment with lead – 11.3%; with manganese – 6.1%, while in the control, the number of white blood cells compared to the total number of formed elements of fish was only 1.2%. Cytometric studies have revealed that there are significant differences in the shape, size, and location of hepatocytes in different experimental fish. The structural components of the liver for histological specimens were stained differentially with different intensities and different colours. The hepatocytes on histological specimens of the liver of the stone moroko exposed to Ni and Mn ions did not have clearly defined boundaries, there was a large number of destroyed cells, which indicates the toxic effect of these heavy metals. The hepatocytes exposed to Pb had the largest area of cells and nuclei and the highest nuclear-cytoplasmic ratio is typical for hepatocytes under the influence of Mn. The nuclei had the largest size and occupied 12.7% of the internal contents of the cells. It is shown that under the impact of the studied concentrations of Mn, Ni and Pb, lactate dehydrogenase activity increased by 1.22, 1.14 and 1.48 times compared to the control, respectively. In contrast, there was a 3.27-fold decrease in succinate dehydrogenase activity under the impact of Mn. Besides that, the activity of the enzyme decreased by 1.48 and 1.68 times under the action of Ni and Pb. Subsequently, we found an increase in the activity of alkaline phosphatase in muscle tissue by 3.25–3.94 times under the influence of the studied toxicants. Muscle protein levels under the impact of Mn were 1.14 times lower than in the control, the most distinct decrease in protein was found under the impact of Ni (1.53 times). The obtained data of physiological and biochemical reactions of the stone moroko to the influence of heavy metals provide an opportunity to predict changes in the species composition of fish fauna under conditions of excessive toxic pollution of ecosystems.

Changes in biomarkers of endothelial function in the blood after intracerebral hemorrhage in rats with type 2 diabetes mellitus

Clinical evidence suggests that type 2 diabetes mellitus can increase the risk of intracerebral hemorrhage and provocation of neurodegeneration. This study was aimed at evaluating biomarkers of glycemic control, lipid profile, oxidative modification of proteins, as well as the functional state of endothelium in Wistar rats with type 2 diabetes mellitus complicated by intracerebral hemorrhage. Experimental type 2 diabetes mellitus was induced by intraperitoneal injection of streptozotocin (65 mg/kg) and nicotinamide (230 mg/kg). The intracerebral hemorrhage was induced by microinjection of sterile saline containing 0.2 IU bacterial collagenase into the striatum. Assessed biomarkers included the area under glycemic curve, glycated hemoglobin, total cholesterol, triglyceride, high-density lipoprotein, advanced glycation end products, markers of oxidative modification of proteins – aldehyde- and ketonephenylhydrazones, and markers of endothelial dysfunction – homocysteine, endothelin-1, von Willebrand factor and asymmetric dimethylarginine in blood serum. Both rats with type 2 diabetes mellitus and rats with intracerebral hemorrhage and diabetes had a significant elevated glycemic control as compared to intact animals. But combined pathology was additionally characterized by an impairment of lipid profile (increased triglyceride level and decreased as total cholesterol and high-density lipoprotein) resulting in a rise in the atherogenic index of plasma. A significant increase in the content of the markers of oxidative modification of proteins was observed in both experimental groups. But the rats with intracerebral hemorrhage and diabetes only had higher levels of advanced glycation end products in comparison with intact animals. The highest levels of endothelin-1, as a biomarker of endothelial dysfunction, were observed in animals with intracerebral hemorrhage and diabetes. Homocysteine and von Willebrand factor were elevated in rats with type 2 diabetes mellitus, while acute intracerebral hemorrhage did not potentiate the further growth in its levels. Such effect was not accompanied by a marked increase of asymmetric dimethylarginine level in blood serum, although there was a clear trend. In conclusion, the development of intracerebral hemorrhage in rats with type 2 diabetes mellitus can intensify the manifestations of oxidative stress, worsen lipid profile, and aggravate endothelial dysfunction. In this case, the pathological process may have the character of a “vicious circle”.

Morphological and physiological-biochemical variability of spore-forming bacteria isolated from the agrocoenosis of winter wheat

From an agrocoenosis of winter wheat (Triticum aestivum L.; phylloplane and rhizosphere of the root system; typical chernozem, soil column measuring up to 40 cm), using the classical microbiological methods, we had isolated soil bacteria and characterized them according to the morphological features as representatives of Gram-positive and spore-forming bacteria of Bacillus sp. genus. In the earing-swelling phase of grain, the screening studies found non-pigmented forms of colonies of bacterial isolates, 19 of which were classified to colonial-morphological diversity of R-type with the diameter of 7 to 13 mm. The analysis of physiological condition of cells of populations of soil isolates revealed technologic specificity according to parameters of spore formation in different conditions and incubation time (up to 48–72 h). We observed 90.0% of free spores in axenic cultures as early as after 72 h of cultivation and no more than 10.0% of prospores in the studied monoisolates with stable morphologic traits. Isolates Н10 and Н45 demonstrated the ability to grow in higher cultivation temperatures (+37…+40 °С). According to environmental рН, isolates were able to grow in рН ranging 4.5–8.0. Differential diagnostic testing revealed that as the source of carbon, with formation of acid, soil isolates used arabinose, xylose, mannitol, glucose, galactose, fructose, maltose, sorbitol, glycerin, dextrin, starch, rhamnose and dulcite (with development of alkaline). There was observed active use of mineral forms of nitrogen: ammonium salt and nitrates, aminoacids and proteins. The isolates hydrolyzed casein, gelatin, starch, and litmus was being reduced in the young during growth in milk with litmus. They also exerted catalase activity and were oxidase-positive. Biochemical testing using API test system determined that the studied isolated bacteria differed by a range of fermentation carbohydrates, reduction of nitrates. In the conditions of submerged fermentation, isolates Н38 and Н40 grew in heightened temperature ranges of cultivation (40 °С) for 48 h (according to fact of spore development). Therefore, according to the key morphologic and biochemical traits, strains Н3, Н10, Н13, Н36, Н38, Н40, Н43, Н45 were similar to such of reference strain B. subtilis 8A, and were identified to Bacillus sp., species B. subtilis.

Oxidative stress in moss Bryum caespiticium (Bryaceae) under the influence of high temperature and light intensity in a technogenically transformed environment

Mosses are pioneer plants in post-technogenic areas. Therefore, the question of adaptive reactions of mosses from these habitats represents a scientific interest. The research is devoted to the study of adaptive changes in the metabolism of the dominant moss species Bryum caespiticium Hedw., collected in the devastated territories of the Novoyavorivsk State Mining and Chemical Enterprise (SMCE) “Sirka (Sulfur)” exposed to hyperthermia and insolation, which cause oxidative stress in plants. The influence of these stressors on the activity and thermal stability of antioxidant enzymes, hydrogen peroxide content, anion radical generation and accumulation of prooxidant components in moss shoots was studied. The activity and thermal stability of peroxidase and superoxide dismutase (SOD) were analysed forB. caespiticium moss from different locations of northern exposure at the sulfur mining dump No 1 in summer and autumn. We established the dependence of the activity of antioxidant enzymes of moss on the intensity of light and temperature on the experimental plots of the dump No 1. In summer, the highest activity and thermal stability rates of peroxidase and SOD were observed. Under the conditions of the experiment in shoots of В. caespiticium from the northern peak of the dump under the influence of 2 hours temperature action (+ 42 ºС) the most significant increase in peroxidase activity was found by 1.78 times and SOD by 1.89 times, as well as increase in its thermal stability by 1.35–1.42 times, respectively. The increase in peroxidase and SOD activity, as well as the increase in their thermal stability caused by hyperthermia were negated by pre-processing with a protein biosynthesis inhibitor cyclohexamide, which may indicate the participation of the protein-synthesizing system in this process. The effect of increasing the thermal stability of enzymes can be considered as a mechanism of adaptation of the protein-synthesizing system to the action of high temperatures. Increase in the activity and thermal stability of antioxidant enzymes is caused primarily by changes in the expression of stress protein genes, which control the synthesis of specific adaptogens and protectors. The obtained results indicate that the extreme conditions of the anthropogenically transformed environment contribute to the development of forms with the highest potential abilities. The mechanism of action of high temperatures is associated with the development of oxidative stress, which is manifested in the intensification of lipid peroxidation and the generation of superoxide anion radical. It was found that temperature stress and high insolation caused an increased generation of superoxide anion radical as the main inducers of protective reactions in the samples of B. caespiticium from the experimental transect of the sulfur mining heap. It is known that the synthesis of Н2О2 occurs under stress and is a signal to start a number of molecular, biochemical and physiological processes of cells, including adaptation of plants to extreme temperatures. It is shown that high temperatures initiate the generation of hydrogen peroxide. Increased reactive oxygen species (ROS) formation, including Н2О2, under the action of extreme temperatures, can cause the activation of signaling systems. Therefore, the increase in the content of Н2О2 as a signaling mediator is a component of the antioxidant protection system. It is determined that adaptive restructuring of the metabolism of the moss В. caespiticium is associated with the accumulation of signaling prooxidant components (diene and triene conjugates and dienketones). The increase in primary lipid peroxidation products, detected by us, under the action of hyperthermia may indicate the intensification of free radical oxidation under adverse climatic conditions in the area of the sulfur production dump, which leads to the intensification of lipid peroxidation processes. The accumulation of radical and molecular lipid peroxidation products are signals for the activation of protective systems, activators of gene expression and processes that lead to increased resistance of plants.

Influence of metal nanocarboxylates and different water supply conditions on efficiency of soybean-rhizobial symbiotic systems

Insufficient water supply is one of the main factors that significantly reduce the activity of nitrogen fixation by legume-rhizobial symbiotic systems. That is why comprehensive research on aspects of their resistance to water stress and the search for scientifically substantiated ways to improve the existing ones and develop modern, competitive technologies of growing legumes in arid conditions are becoming especially relevant. The aim of the work was to investigate the processes of formation and functioning of soybean-rhizobial symbiotic systems developed under conditions of different water supply and influence of nanocarboxylates of cobalt, ferum, germanium, chromium, сuprum and molybdenum. The nanoparticles of specified metal nanocarboxylates were used as components of the inoculation suspension of rhizobia of Tn5 mutant B1-20 for soybean seed treatment. A model drought lasting 14 days was created by controlled irrigation. Microbiological and physiological research methods were used in the study. We determined that insufficient water supply caused a significant decrease in the nodulation potential of rhizobia and the intensity of molecular nitrogen fixation by symbiotic systems formed with the participation of soybean plants and nodule bacteria without adding these metal nanocarboxylates to the inoculation suspension. Application of most of the metal nanocarboxylates used as components of the inoculation suspension mitigated the negative impact of stress on the investigated parameters. The study revealed the stimulating effect of cobaltnanocarboxylate on the activity of molecular nitrogen fixation, which was more pronounced in the conditions of insufficient water supply. Symbiotic soybean systems formed with the participation of nodule bacteria containing germaniumcarboxylate nanoparticles were proved to be the least sensitive to the negative impact of insufficient water supply. This was indicated by high rates of nodulation and nitrogen-fixing activity compared with other studied symbiotic systems. We confirmed that the addition of chromium nanocarboxylate to the inoculation suspension of rhizobia provided the highest rates of nodulation and nitrogen-fixing activity of soybean root nodules under optimal growing conditions and, at the same time, had no noticeable positive effect under water stress. We determined that сuprum and molybdenum nanocarboxylates, as components of the inoculation suspension, regardless of the water supply level, had a less notable positive effect on the processes of nodule formation and nitrogen fixation, and in some cases even led to a decrease in the investigated values for control plants. Thus, the study demonstrated that the use of germanium, cobalt and ferum nanocarboxylates as components of the bacterial suspension helped to increase the adaptation of the formed legume-rhizobial symbiotic systems to water stress, as evidenced by the maximum indexes of nodulation and molecular nitrogen fixation in the context of insufficient water supply and recovery of their level to optimal after the stress influence had ended. Based on the results, it was concluded that inoculation of seeds by the complex bacterial preparations made on the basis of Bradyrhizobium japonicum B1-20 with a content of germanium, cobalt and ferum nanocarboxylates in the concentration of 1:1000 can become one of the important means in soybean growing technologies of increasing the nitrogen-fixing potential and resistance of plants to insufficient water supply.

Specifics of the morphological identification of the pathogen of passaluariasis of rabbits

Passalurus ambiguus (Nematoda, Oxyuroidae) is one of the most common parasites of digestive tract of domestic and wild rabbits and rodents. Passalurus spp. are cosmopolitan, and the mature nematodes are localized in the appendages and large intestine of animals. The passaluariasis infections remain quite relevant in modern rabbit breeding, because the disease is highly contagious, with the possibility of unlimited spread inducing significant economic losses. The aim of the study was to determine the identifying morphological species characteristics of P. ambiguus nematodes, which were isolated from the colon of domestic rabbits (Oryctolagus cuniculus domesticus). The identifying characters of Passalurus include morphological features, namely the presence of a spherical bulb in the esophagus, and the presence of three teeth in the oral capsule. Males of P. ambiguus have a characteristic structure of the tail end, namely the presence of papillary protrusions and a massive narrowed part with a spicule handle, and five papillae around the cloaca. Twenty-eight metric indicators are proposed that characterize the size of the body, oral capsule, esophagus in different parts of it, the tail end, spicules, the location of the cloaca. Two different morphotypes of mature females of P. ambiguus were revealed – short-tailed and long-tailed, which were larger by 19 indicators (by 3.2–43.6%). The morphotypes of P. ambiguus females are characterized; the length of the tail, body length and width in different areas, depth of the oral capsule, esophageal size and its structural elements, the location of the vulva and nerve ring, the width of the caudal process. At the same time, the parameters of esophageal bulb length and caudal process in short-tailed females were higher (by 13.9% and 11.9%, respectively) than in the long-tailed females. A total of 25 morphometric parameters were proposed for the identification of P. ambiguus females. The obtained data indicate the feasibility of further research on the causes of occurrence of such morphotypes in P. ambiguus females and the need to take into account the existence of such forms of females for their differential diagnosis taking into account their proposed morphometric parameters.

Effects of humate and neodymium on phytochemical levels in kale at different ontogenetic stages

The increased global mortality caused by the rise in chronic diseases can be reduced by the consumption of antioxidant-rich foods. Leafy green kale (Brassica oleracea var. sabellica) has high nutritional value due to its high antioxidant content and its consumption is highly correlated with a reduced risk of developing some chronic diseases. The paper focuses on examining the effects of two different mineral nutrients – neodymium and potassium humate – on the accumulation of flavonoids, vitamin C, phenolic compounds, carotenoids and malondialdehyde (MDA) content in field-grown leafy green kale. Leaves from treated plants were collected at three stages of maturity –14 weeks, 18 weeks, and 22 weeks and subjected to spectrophotometric analysis. The results showed that the stimulatory effect of both mineral nutrients on the phytochemicals varied at the different growth stages - the highest significant effect of neodymium has been mostly observed at week 18 with high flavonoid, vitamin C, and carotenoid levels. Phenolic compounds for this variant are the same as in the control samples, while the level of malondialdehyde was reduced by 21.8%, signifying increased antioxidant activity. The rare earth element primarily maintained some phytochemical content at weeks 14 and 22. Following soil treatment with potassium humate, the levels of studied phytochemical compounds were either maintained or elevated at weeks 14, 18, and 22. Humic acid exerted the greatest decrease in malondialdehyde content in kale at week 14, indicating a reduction in the lipid peroxidation process in leaves. Accordingly, the harvest date of leafy green kale (Brassica oleracea var. sabellica) should depend on the type of mineral nutrient applied. The obtained results provide information that may be relevant to the production of functional varieties and enhance the nutritional and possibly the economic value of kale.

Differential diagnosis of Dirofilaria immitis nematodes (Nematoda, Onchocercidae)

Heartworm disease is a widespread anthropozoonotic disease of carnivorous animals, as well as humans. It is caused by nematodes belonging to the suborder Filariata, family Onchocercidae, genus Dirofilaria. There are about 26 species of heartworms in nature, the most common and pathogenic species in dogs and cats in most countries is Dirofilaria immitis Leidy, 1856. Mature helminths parasitize in the right ventricle and pulmonary arteries, large veins of animals and cause heart and vascular disorders, and death. Therefore, the aim of the study was to investigate the features of morphological and metric structure of adult nematodes of D. immitis isolated from the heart of dogs. Morphological studies have shown that in males the most characteristic differential features are the presence of two unequal spicules, specifically positioned relative to each other, as well as well-defined preanal and less pronounced adanal and postanal papillae. In female heartworms, the characteristic morphological features are the shape and location of the vulva. There is a difference in the structure of the esophagus in males and females. In females, the anterior and posterior parts of the esophagus are well expressed, with enlargements, in males these divisions are not pronounced. To increase the efficiency of species identification of D. immitis nematodes, it is proposed to use metric parameters that characterize the overall body size, body and width of esophagus in different areas, length of esophagus, and the location of the nerve ring. In males, 11 indicators are also suggested that characterize the size of the spicules and the location of the cloaca. In females, seven additional parameters are pointed out that characterize the location of the vulva, anus and body width in these areas. The obtained data expand the already existing data on the peculiarities of the morphological structure of parasitic nematodes of the species D. immitis and their identification.

GC-MS analysis of cuticular waxes and evaluation of antioxidant and antimicrobial activity of Chaenomeles cathayensis and Ch. × californica fruits

Fruit extracts of the Chaenomeles species are a rich source of compounds having health-promoting properties, while their distribution between the species and cultivars varies significantly depending on both genotype and environmental threats. This study aimed at discovering antioxidant and antimicrobial potential of the secondary metabolites of fruit and waxes of fruit cuticular of introduced Ch. cathayensis and Ch. × californica plants. The sum of detected polyphenols in the isopropanolic fruit extracts varied slightly between the species, while significant excesses in indices were seen for both species peel extracts as compared to pulp extracts. Antimicrobial assays carried out by disc diffusion method showed notable activity of the fruit peel and pulp extracts of both species against all tested Gram-negative and Gram-positive bacterial strains, and two Candida strains as well. Pseudomonas aeruginosa strain was the most resistant to the action of both fruit extracts, especially peel extracts of Ch. cathayensis fruits. As identified by gas chromatography-mass spectrometry (GC-MS) assays, chloroformic extracts from the fruits of cuticular waxes of Ch. cathayensis and Ch. × californica contained six prevailing fractions: aldehydes, alkanes, alcohols, esters, fatty acids and various terpenoids. The predominant compounds were tetrapentacontane (21.8% of total amount) and heptacosanal (23.1% of total), respectively in the cuticular waxes of Ch. cathayensis and Ch. × californica. Cinnamaldehyde, cis-9-hexadecenal, hexadecanoic acid, oleic acid, olean-12-ene-3,28-diol (3. beta), lupeol, diisooctyl phthalate, 9-octadecenoic acid, 1,2,3-propanetriyl ester, 1,3,12-nonadecatriene-5,14-diol and some other identified compounds are well-known for their bioactivity, indicating the feasibility of studying the antimicrobial potential of plant fruits.