Aim. The aim of this study was to investigate how viral infections, which severely impair plant growth and productivity, affect reactive oxygen species (ROS) accumulation, and to evaluate the roles of both enzymatic antioxidant defenses and non-enzymatic low-molecular weight antioxidants, including ascorbate (ASA), glutathione (-L-glutamyl-L-cysteinyl-glycine = GSH), and tocopherols (TOCs), in mitigating virus-induced oxidative stress in plants. Methods. In the present study, a total of 45 leaf samples were collected from mature 10-year-old vines of the cultivars 'Gara Shani' (red) and 'Ag Shani' (white). Sampling was conducted in three vineyards, where five vines were selected per vineyard, and three physiologically mature leaves were collected from each vine, resulting in 15 leaves per vineyard and 45 leaves in total (3 vineyards × 5 vines × 3 leaves). In each vineyard, one vine without visible symptoms was designated as a healthy control, while the remaining vines exhibited typical viral symptoms such as reddening of leaf margins and petioles, reduced vigor, leaf rolling, yellowing, and curling. Consequently, 36 leaves were obtained from symptomatic vines and 9 leaves from asymptomatic (healthy) vines. Grapevine leafroll-associated virus 3 (GLRaV-3) infection was confirmed in grapevine samples using AgriStrip rapid assay, double-antibody sandwich ELISA (DAS-ELISA), and reverse transcription-polymerase chain reaction (RT-PCR). Subsequently, virus-induced changes in non-enzymatic antioxidants were quantified by spectrophotometric determination of ascorbate and glutathione, tocopherols via the Emmerie-Engel reaction, and lipid peroxidation through MDA content using the thiobarbituric acid reactive substances (TBARS) assay. Enzymatic antioxidant activities, including ascorbate peroxidase and catalase, were measured spectrophotometrically, and soluble protein content was determined by the Sedmak method. Results. Viral screening using AgriStrip assays and DAS-ELISA, followed by RT-PCR targeting the coat protein gene, confirmed GLRaV-3 infection in 13 samples (approximately 36.1%), while symptomless vines tested negative, demonstrating consistency between visual assessment and molecular diagnostics. GLRaV-3 infection resulted in significant increases in non-enzymatic antioxidant levels compared with healthy controls. Tocopherol (TOC) content increased by 18-73% across infected samples 2-6, whereas sample 1 showed no significant change. Glutathione (GSH) levels were elevated by 20-60% in infected samples 1, 2, 4, 5, and 6, while sample 3 showing no significant difference. Ascorbic acid (ASA) content increased by 10-23% in infected samples 1, 3, 5, and 6, with sample 2 and 4 showing no significant difference. Lipid peroxidation, as indicated by malondialdehyde (MDA) content, was also significantly enhanced in infected leaves. MDA levels increased approximately 1.5-2.6-fold in samples 1, 2, 4, 5, and 6 compared with healthy controls, reaching absolute values of 0.85-1.75 µmol g-1 FW versus 0.45-0.9 µmol g-1 FW in controls. In parallel, the activities of key antioxidant enzymes were elevated: catalase (CAT) activity increased by 25-48%, and ascorbate peroxidase (APX) activity increased by 30-55% in infected leaves compared with corresponding controls. Conclusions. Surveys conducted in Salyan vineyards confirmed GLRaV-3 infection supporting the link between observed symptoms and biochemical oxidative stress responses. These biochemical changes demonstrate that GLRaV-3 infection triggers oxidative stress and concurrently activates both enzymatic and non-enzymatic antioxidant defense mechanisms in grapevine leaves. The coordinated increase in ASA, GSH, TOC, CAT, and APX, alongside elevated MDA, indicates a complex redox adjustment aimed at mitigating virus-induced oxidative damage. ```
Aim. The aim of this study was to investigate how viral infections, which severely impair plant growth and productivity, affect reactive oxygen species (ROS) accumulation, and to evaluate the roles of both enzymatic antioxidant defenses and non-enzymatic low-molecular weight antioxidants, including ascorbate (ASA), glutathione(-L-glutamyl-L-cysteinyl-glycine = GSH), and tocopherols (TOCs), in mitigating virus-induced oxidative stress in plants. Metods. In the present study, a total of 45 leaf samples were collected from mature 10-year-old vines of the cultivars `Gara Shani' (red) and `Ag Shani' (white). Sampling was conducted in three vineyards, where five vines were selected per vineyard, and three physiologically mature leaves were collected from each vine, resulting in 15 leaves per vineyard and 45 leaves in total (3 vineyards X 5 vines X 3 leaves). In each vineyard, one vine without visible symptoms was designated as a healthy control, while the remaining vines exhibited typical viral symptoms such as reddening of leaf margins and petioles, reduced vigor, leaf rolling, yellowing, and curling. Consequently, 36 leaves were obtained from symptomatic vines and 9 leaves from asymptomatic (healthy) vines. Grapevine leafroll-associated virus 3 (GLRaV-3) infection was confirmed in grapevine samples using AgriStrip rapid assay, double-antibody sandwich ELISA (DAS-ELISA), and reverse transcription-polymerase chain reaction (RT-PCR). Subsequently, virus-induced changes in non-enzymatic antioxidants were quantified by spectrophotometric determination of ascorbate and glutathione, tocopherols via the Emmerie-Engel reaction, and lipid peroxidation through MDA content using the thiobarbituric acid reactive substances (TBARS) assay. Enzymatic antioxidant activities, including ascorbate peroxidase and catalase, were measured spectrophotometrically, and soluble protein content was determined by the Sedmak method. Results. Viral screening using AgriStrip assays and DAS-ELISA, followed by RT-PCR targeting the coat protein gene, confirmed GLRaV-3 infection in 13 samples (approximately 36,1%), while symptomless vines tested negative, demonstrating consistency between visual assessment and molecular diagnostics. GLRaV-3 infection resulted in significant increases in non-enzymatic antioxidant levels compared with healthy controls. Tocopherol (TOC) content increased by 18-73% across infected samples 2-6, whereas sample 1 showed no significant change. Glutathione (GSH) levels were elevated by 20-60% in infected samples 1, 2, 4, 5, and 6, while sample 3 showing no significant difference. Ascorbic acid (ASA) content increased by 10-23% in infected samples 1, 3, 5, and 6, with sample 2 and 4 showing no significant difference. Lipid peroxidation, as indicated by malondialdehyde (MDA) content, was also significantly enhanced in infected leaves. MDA levels increased approximately 1.5-2.6-fold in samples 1, 2, 4, 5, and 6 compared with healthy controls, reaching absolute values of 0.85-1.75 & micro;mol g-1 FW versus 0.45-0.9 & micro;mol g-1 FW in controls. In parallel, the activities of key antioxidant enzymes were elevated: catalase (CAT) activity increased by 25-48%, and ascorbate peroxidase (APX) activity increased by 30-55% in infected leaves compared with corresponding controls. Conclusions. Surveys conducted in Salyan vineyards confirmed GLRaV-3 infection supporting the link between observed symptoms and biochemical oxidative stress responses. These biochemical changes demonstrate that GLRaV-3 infection triggers oxidative stress and concurrently activates both enzymatic and non-enzymatic antioxidant defense mechanisms in grapevine leaves. The coordinated increase in ASA, GSH, TOC, CAT, and APX, alongside elevated MDA, indicates a complex redox adjustment aimed at mitigating virus-induced oxidative damage.
Aim. In Cote d'Ivoire, cocoa pod brown rot caused by Phytophthora megakarya represents a major constraint to sustainable cocoa production due to the severe yield losses it induces. This study aimed to evaluate the antifungal activity of four Trichoderma species (T. harzianum, T. asperellum, T. virens and T. hamatum) against P. megakarya under in vitro and in vivo conditions using detached cocoa pods. Methods. In vitro assays were conducted using a dual culture technique on PDA medium to assess the inhibition of mycelial growth of the pathogen. In vivo tests consisted of preventive treatments followed by aggressive and non-aggressive inoculations on healthy cocoa pods. Results. The in vitro results showed a significant inhibition of P. megakarya mycelial growth by all Trichoderma species, with inhibition rates ranging from 77.27% to 89.27%. Trichoderma virens exhibited the highest antagonistic activity. Under in vivo conditions, T. virens and T. harzianum significantly reduced the severity and spread of cocoa pod brown rot, with inhibition rates exceeding 80% depending on the inoculation method. Conclusions. The in vitro and in vivo assays showed that all tested species exert a significant inhibitory effect on the development of the pathogen, although the efficacy varies according to the Trichoderma species. Among the evaluated species, Trichoderma virens stood out for its strongest ability to inhibit the mycelial growth of P. megakarya and for a marked reduction in disease severity on cocoa pods under both aggressive and mild inoculation conditions. These results confirm the strong potential of Trichoderma species, particularly T. virens, as effective biocontrol agents against P. megakarya and support their integration into sustainable management strategies for cocoa pod brown rot, however, further field studies are necessary.
Aim. To investigate the genetic structure of populations of the Ukrainian Red-and-White Dairy cattle (URW) and its crossbreeds based on SNP g.10153G>A at SCD1 locus and to evaluate the association of this polymorphism with traits of dairy performance and the fat and protein content in milk. Methods. The genetic structure of two groups of cows (purebred URW, n = 43; URM & times; M crossbreeds, n = 41) was studied by g.10153G>A polymorphism of SCD1 gene using the PCR-RFLP method. The performance was assessed using the indicators from the first completed lactation. The correspondence of the distribution of quantitative traits to the normal law was tested using the Shapiro-Wilk test. For normally distributed traits (milk yield), one-way analysis of variance (ANOVA) with Tukey's post-hoc test was used; for non-normally distributed traits (fat and protein content), the Kruskal-Wallis test with Dunn's post-hoc test adjusted for Bonferroni correction was used. Results. Certain intergroup differences in allele distribution were observed: in purebred animals, a slight predominance of allele G was observed (0.535 versus 0.465 for allele A), whereas in crossbreeds, a relative increase in the frequency of allele A was noted (0.524 versus 0.476 for allele G). In the crossbreed group, an increased frequency of the heterozygous AG genotype (0.512) was also observed. For milk yield indicators, opposite genotype-associated trends were identified between groups: in purebred animals, the highest mean values were observed in homozygous AA individuals, whereas in crossbreed animals, they were observed in heterozygous AG individuals; however, within individual groups, no statistically significant differences between genotypes were found (p > 0.05). No statistically significant associations with genotype were found for fat and protein content (p > 0.05). Conclusions. The results obtained indicate the presence of certain genetic differentiation between the study groups at the SCD1 locus and a possible dependence of the manifestations of the associations on the genetic background of the population. At the same time, no statistically confirmed effects of polymorphism on milk yield, fat content, or protein content were found. Further studies using larger sample sizes are needed to clarify the nature of the identified trends.
Aim: To estimate the seroprevalence of bovine brucellosis (bBR) and identify risk factors associated with sero-positivity to this disease. An additional aim was to evaluate heifer-related risk factors for abortion. Methods: A total of 3,848 Holstein heifers with a single lactation from a large commercial dairy herd in northern Mexico (25 degrees N) participated in this study. Multiple regression models were employed to analyze risk factors for brucellosis seropositivity rate and to calculate odds ratios, quantifying the strength of the risks. Results: The seroprevalence of bBR was 30% (95% CI = 29-32%) for lactating heifers (card test). Heifers weighing more than 680 kg at calving had half the risk of testing positive for bBR compared to those weighing less than 680 kg (22.7% vs. 36.1%). The risk of being seropositive for bBR increased in heifers with retained placenta compared to those without this reproductive disorder (OR = 2.46; 47.5% vs. 26.3%). Heifers vaccinated with two doses of a commercial brucellosis vaccine had 2.2 times higher odds of being seropositive for bBR compared with those vaccinated only once (P < 0.01). Seropositive heifers for bBR were 3.1 times more likely to abort than seronegative heifers (38.5% vs. 16.2%; P < 0.01). Heifers with a body condition score (BCS) of 3.5 or higher at calving were less likely to abort than heifers with BCS < 3.5. Conclusions: Brucellosis control programs should prioritize age at first calving, placenta retention, and the number of RB51 vaccinations administered to Holstein heifers.
Aim. To substantiate the approach towards the identification of zones, vulnerable to nitrate accumulation, based on the data of mineral nitrogen content in soil according to the requirements of the Nitrate Directive. Methods. The study was conducted on arable soils of humus-accumulative genesis on six land plots in the Forest-Steppe of Ukraine with the rate of applying nitrogen-containing fertilizers from 0 to 170 kg/ha. The content of mineral nitrogen (Nmin), nitrates and productive moisture reserves was determined layer-wise down to the depth of 120 cm, in dynamics during the vegetative periods of 2021, 2024, and 2025. Crops: grain corn, winter wheat, spring barley, oil flax. Results. A considerable variability in the content of Nmin and nitrates proper was determined in soil depending on terms, ways, and norms of applying nitrogen-containing fertilizers. The maximal concentrations were observed in the arable soil layer immediately after the application of nitrogen fertilizers, but these parameters gradually decreased with time and depth. In June-July, the highest accumulation of nitrates in deep soil layers was noted when, at the norm of the application of fertilizers of over N100, the content of nitric nitrogen at the depth from 80 to 120 cm was from 15.6 to 35.8 mg/kg. The accumulation of nitrates in deep layers of soil decreases in the course of the vegetation period, and at the time of harvesting, the content of nitric nitrogen at the depth of 80-120 cm did not exceed 10 mg/kg in a year with normal moisturization conditions and 4 mg/kg in a dry year. At the norm of applying fertilizers up to 100 kg/ha, the accumulation of Nmin and nitrates in deep soil layers is minimal. Conclusions. The use of Nmin as an indicator corresponds to the requirements of the Nitrate Directive regarding the identification of zones, vulnerable to the accumulation of nitrates. It is reasonable to take measurements after harvesting the crops. The application of nitrogen-containing fertilizers for the crops of rotations on arable soils of the humus-accumulative genesis in the plains of Ukraine with non-flushing or periodically flushing type of the water regime at the norm, not exceeding 100 kg/ha, does not pose any threat of leaching mineral nitrogen and its nitrate forms from soil and further pollution of the underground waters.
Spelt wheat (Triticum aestivum ssp. spelta (L.) Thell.) is a type of hulled wheat whose genome is the same as that of hexaploid common wheat (T. aestivum L.) and is characterized by many valuable properties, including plant resistance to pathogens, better adaptability to adverse environmental factors, increased protein content in the grain, and nutritional value of the grain. In recent years, the popularity and use of spelt in the world, and in Ukraine in particular, has been steadily growing, which is associated with the development of organic farming, as well as the need for high-quality food products provided by this crop. In this regard, the purpose of this review was to present the current state of research into the biological properties of spelt wheat, as well as the prospects for its use in breeding. The current hypotheses of the origin of this crop, its distribution in the world and Ukraine, as well as the biological characteristics of this cereal are briefly considered. The genetic features of spelt are highlighted in detail, in particular the genes of morphological traits, vernalization and photoperiod, the variability of waxy genes. The polymorphism in the genes of storage proteins and the prevalence of their alleles, which can be used as marker systems and a source of enriching the common wheat gene pool, are described. The modern data on the quality of spelt grain, including its physical properties, chemical composition, functional-technological and baking properties of flour are presented. The physiological effect of spelt consumption is highlighted. This cereal is considered as a source of useful genes for common wheat breeding. The information on modern strategies and new scientific directions of genetic improvement of this crop is presented.
The aim of the study was to investigate the impact of a paracetamol (PCM)-and additive-containing pharmaceutical (Infulgan) on growth characteristics of the seeds and seedlings of Triticum aestivum L. and Lepidium sativum L., phytotoxicity for L. sativum and on microbial contamination of seeds for T. aestivum. Methods. Winter soft wheat (T. aestivum) variety Bogdana and garden cress (L. sativum) variety Afrodita were used as test plants in laboratory conditions (Problematic Research Laboratory of "Ecological Biochemistry, Ichthyology and Biocorrosion" of T.H. Shevchenko National University "Chernihiv Colehium"). Aqueous solutions of Infulgan were used, which contained from 0.1x10-5 (10 mg/L) to 0.2% w/v (2,000 mg/L) PCM. In a growth test, the germination energy (%), germination (%) of T. aestivum and L. sativum seeds (3rd day), and some biometric and morphometric indices of seedlings were evaluated: T. aestivum: coleoptile length, number of roots on the 7th day; L. sativum - length of roots and shoot on the 5th day, simplified vitality index, relative root growth percentage. Phytotoxic indices were estimated for L. sativum only. The seed contamination incidence percentage (CIP) was also estimated. The results were analyzed using mathematical and statistical methods. Results. It was established that: 1) germination energy and seed germination rate did not change in all treatments with all concentrations of PCM investigated, when using the pharmaceutical Infulgan; 2) the growth indices of wheat and garden cress were statistically significantly impaired by a solution containing 0.05-0.2% PCM. Moreover, wheat seed was affected considerably by unde-termined microbial contamination. A statistically significant decrease (1.2-2.5-fold) in the length of the wheat coleoptile compared to the control was recorded, along with an increase in the number of roots per seed (from 5.9 +/- 0.1 to 6.0 +/- 0.4 compared to 4.4 +/- 0.2 in the control). A significant increase in wheat root formation (5.2 +/- 0.2 roots per seed) was also recorded for PCM at 0.025%. PCM concentrations of 0.025%, 0.002%, 0.25x10-4 % and 0.1x10-5 % did not negatively impact plant growth indices. Conclusions. The germination energy and seed germination rate did not change under the influence of PCM as formulated in the pharmaceutical Infulgan at any concentration tested, however, the biometric indices of seedlings changed significantly. A substantial toxicity of Infulgan solutions was observed for L. sativum, at concentrations higher than 0.025%. Infulgan at a 0.05-0.2% PCM concentration enhanced root formation with 36%, where not only PCM, but also the excipients of the drug could have played a (so far unexplored) role. In wheat seed with higher PCM concentrations a substantial (so far undefined) microbial contamination of 77-100% was observed. Therefore, the ecotoxicity of PCM and its formulations should be assessed more comprehensively, taking PCM and its excipients (as formulation and as separate compounds) into account. At present known PCM concentrations characteristic of wastewater and environment appear not to pose a risk to the germination and growth of wheat and water cress.
Aim. To forecast the monthly percentage of Holstein cows and heifers at a high-input dairy farm experiencing retained placenta (RP), puerperal metritis (PM), and clinical ketosis (CK). Methods. An autoregressive integrated moving average (ARIMA) model was employed to predict future monthly cases of these diseases using time series data. These puerperal diseases were observed on a single dairy farm with 2560 to 3300 milking cows over seven years, from 2014 to 2020. Results. The highest predicted RP incidence in cows was in May (11.3%; 95% CI=6.3-16.4), while the lowest was in November (5.4%; 95% CI=0.5-10.4). For heifers, the peak predicted RP occurrence was in August (20.6%; 95% Cl=11.0-23.1), and the lowest was in December (10.5%; 95% CI=7.9-13.0). The highest projected CK occurrence in cows was in June (3.0%; 95% CI=1.8-4.3) and the lowest was in November (1.1%; 95% CI=-0.1-2.4). For heifers, CK was most likely in May (2.7%; 95% CI=0.9-4.5) and least in December (0.7%; 95% CI=-1.1-2.5). Conclusions. Both cows and heifers showed an increasing trend in RP, PM, and CK during summer months; ARIMA models effectively tracked disease trends throughout the year and can aid in health management decisions for dairy cows.
Aim. To investigate the effect of using different time parameters of starting in vitro activation of oocyte maturation (activation onset) on the viability and further development of oocyte-cumulus complexes (OCC) of cows, the efficiency of oocyte fertilization, and the production and further in vitro development of embryos. Methods. Biotechnological, embryological, morphological, and cytogenetic methods, as well as methods of variation statistics were used in the research. The ovaries of the Ukrainian black-and-white dairy cows (n = 28) were kept at the temperature of +38 degrees & Scy; in the physiological solution. The OCC were obtained by incising visible antral follicles with a blade. The period from the moment of incising the antral follicles to placing the OCC for cultivation was 30-40 min in all the groups. The embryos with more than 16 cells were classified as morulae and the embryos with blastocoel - as blastocysts. All the experimental groups were analyzed in three repeats (n = 3). The statistical processing of the data was conducted using the reliability criterion chi 2, and the diffe-rences were deemed statistically significant at p < 0.05. Results. The study was conducted from 2021 to 2024. The effect of the activation procedure and different terms of its start on the biological completeness and further development of cow oocytes of Ukrainian black-and-white dairy breed (n-28) after meiotic maturation in vitro was studied. The coefficient and index of the fragmentation of the obtained in vitro embryos were analyzed for different time parameters of the activation onset. It was shown that the efficiency of obtaining bovine cattle embryos in vitro and their further development depended on the time of activation onset. The least number of gametes at the metaphase-2 stage (48. 8%) and the most cells with degenerated chromatin (44.0%) were obtained when using the most delayed time of activation onset of in vitro maturation of oocytes (>= 8 hours). In case of activation 2, 4, 6, and 8 hours after the start of cultivation, the rate of morula-blastocyst formation was 11.1, 9.0, 2.9, and 0.0%, respectively. Conclusions. With an increase in the time interval between the start of culture and the activation of oocytes up to 8 hours, the number of the formed in vitro bovine embryos decreased.
Aim. Detection of spatial and temporal specificities of the frequency, duration, intensity, and magnitude of cold waves in Ukraine and determination of tendencies in their changes in 1981-2020. Methods. The study used modern climatological and statistical tools, including the methods of objective identification of extreme events based on daily percentiles of minimum air temperature, probability distribution analysis, and regression approaches to assess the dynamics of changes in indicators. The climate and geographical analysis of seasonal variability of indicators allowed for the detection of their spatial and temporal characteristics in different agroclimatic zones of Ukraine. Results. For the first time, a systematic analysis of cold waves in 1981-2020 was conducted for the entire territory of Ukraine using a unified methodology based on the threshold values of the 32nd, 5th and 0.3rd percentiles of daily minimum air temperatures at 187 meteorological stations. Based on these indicators, a classification of cold waves of varying intensity was proposed: strong (32nd-5th percentile), very strong (5th-0.3rd percentile), and extreme (below 0.3rd percentile). It has been established that an average of 16-17 cold waves are observed in Ukraine each year, 84% of which are strong, 14% are very strong, and about 1% are extreme. A set of spatial and temporal patterns of frequency, duration, intensity, and magnitude of cold waves has been determined for all agroclimatic zones, which allows for a highly detailed characterisation of climatic risks. It has been established that the most intense and powerful cold waves are characteristic of the eastern regions (especially Luhansk, Sumy, and Kharkiv), where the minimum temperature during very strong cold waves in the winter season can drop to -20°C and below, and the magnitude of such waves can reach maximum values. The least frequency and poorly manifested cold waves are observed in western regions, in particular in the Chernivtsi region. Winter cold spells are the most intense and last the longest, while spring cold waves pose the greatest risks to agriculture. Statistically significant trends have been identified in the decrease in the frequency and intensity of cold waves in Ukraine, which correlate with current manifestations of global warming in the mid-latitude of the Northern Hemisphere. A significant weakening of cold waves in Ukraine has been observed over the last four decades. A statistically significant decrease in the recurrence of both short-term (2-5 days) and long-term (≥6 days) cold waves has been recorded throughout the country. At the same time, the decrease in long-term cold spells is more intense and more uniform in space, indicating a weakening of persistent advective intrusions of cold air. Regional anomalies in the change in cold wave characteristics have been identified, in particular, an increase in short-term spring cold spells in the Eastern Forest-Steppe, Steppe zone, and some areas of Polissia and Prykarpattia. This is a new key finding that contradicts the general trend towards weaker cold waves. It has been shown that extreme cold waves have different dynamics than strong and very strong ones - their frequency does not change significantly, and their spatial distribution is clearly associated with local and regional circulation features. Conclusions. Cold waves remain a frequent phenomenon in Ukraine, despite the general warming of the climate. In 1981-2020, Ukraine experienced a statistically significant decrease in the frequency of cold waves, especially in summer and winter, which is consistent with trends in the mid-latitude of the Northern Hemisphere. The number of long-term and short-term cold spells is decreasing, with the frequency of prolonged (≥6 days) cold waves decreasing most rapidly, indicating a weakening of persistent Arctic air intrusions. The intensity and magnitude of cold waves tend to decrease, most noticeably in spring and autumn. Winter and summer changes are regionally heterogeneous, with local areas of intensified cold spells in eastern Ukraine. The greatest risks remain during the transitional seasons, especially in spring, when there is an increase in the recurrence of short-term but sharp cold spells in a number of regions (Eastern Forest-Steppe, Northern and Southern Steppe). This poses a threat to the early stages of crop development. Extreme cold waves in Ukraine are rare and do not change significantly in their frequency, but remain the most dangerous ones in terms of consequences. The most vulnerable regions are Polissia, Forest-Steppe, and the eastern regions. The identified spatial and temporal trends indicate a milder climate regime during the cold season, but high variability and local risks of extreme cold spells remain.
Aim. To study how variations in the antioxidant activity of diets formed exclusively from basic ingredients with different natural antioxidant potential affect the antioxidant status, oxidative stress indicators, biochemical and hematological blood indices, as well as the main productivity indicators of laying hens. Methods. The experiment, conducted from December 2023 to June 2024 at the experimental farm of the State Poultry Research Sta-tion of the Institute of Animal Husbandry of the NAAS (Kharkiv region), involved the use of laying hens of the Birkivska barvysta breed (2 similar groups of 40 birds each), which were kept in group cages. The birds were fed isonutrient diets with the same levels of crude protein (17%) and metabolizable energy (270 kcal/100 g), but contrasting in terms of total antioxidant activity: 12.55 and 23.34 mg/g as recalculated per ascorbic acid. The antioxidant activity of the feeds was modeled based on the actual values of the antioxidant activity of the ingredients (corn, wheat, bran, soybean, and sunflower meal, yeast), which were previously determined by the authors' titrimetric method (Kotyk et al., 2023), expressed as recalculated per ascorbic acid (mg/g). The dura-tion of feeding was 30 weeks. At the end of the experiment, the following indicators of antioxidant status and oxidative stress were determined in hens using common methods: the antioxidant activity of serum, egg white and yolk, the activity of superoxide dismutase and catalase, the level of malondialdehyde in serum; metabolic and hematological indices - total protein, total lipids, cholesterol, alanine aminotransferase, aspartate aminotransferase, hemoglobin, erythrocytes, leukocytes; the markers of non-specific resistance - phagocytic activity of neutrophils and bactericidal activity of serum and egg white regarding Escherichia coli. The antioxidant ac-tivity of excrements was additionally analyzed. The productivity was assessed by egg laying intensity, average egg weight, and survival of the hens; the incubation properties were evaluated by egg hatchability. Results. Higher antioxidant activity of the diet was accompanied by a significant improvement in antioxidant indicators: the antioxidant activity of serum increased by 12.1%, that of egg white by 19.3%, of yolk by 32.0%, superoxide dismutase and catalase activity by 7.4% and 5.5%, respectively, while the level of malondialdehyde in serum decreased by 11.0%. At the metabolic level, an increase in total blood serum protein (by 12.0%) was detected with a simultaneous decrease in total lipids (by 17.4%) and cholesterol (by 14.5%) without changes in the activity of liver enzymes; hematological indices (hemoglobin, erythrocytes, leukocytes) remained within the physiologi-cal norm and did not differ between groups. Among the indicators of non-specific resistance, an increase in the bactericidal activity of egg white (by 3.3%) was noted with unchanged phagocytic activity of neutrophils and bactericidal activity of serum. The antioxidant activity of excrements was higher for the birds that consumed a diet with higher antioxidant activity. The productivity indicators did not differ significantly (egg-laying in-tensity - 52.8% and 52.6%; average egg weight - 58.8 and 59.0 g; survival - 95.6% and 100.0%), and egg hatchability was 90.7% and 91.8% in groups with lower and higher antioxidant activity of the diet, respectively.Conclusions. The optimal combination of ingredients containing natural antioxidants provides an increase in the antioxidant status of laying hens and egg components, a decrease in the intensity of lipid peroxidation, and an improvement in individual metabolic indices without any deterioration in the productivity and incubation indicators. This approach is an effective strategy for increasing the antioxidant protection of both hens and products obtained from them, which is especially important during periods of higher oxidative stress (heat stress, vaccinations, technological stressors).
Aim. To investigate the impact of weather conditions in the Forest-Steppe of Ukraine and genetic specificities of cultivars on the content of vitamin C, polyphenols, flavonoids, and anthocyanins in strawberry fruits. Methods. The study was conducted in 2023-2025; strawberry fruits of 7 cultivars of different ripening periods were selected in the experimental plots of the Institute of Horticulture of the NAAS of Ukraine. The weather indices of the vegetation period prior to collecting strawberry fruits were taken from the meteorological station Vantage Pro2 Plus, located in the experimental plots of the strawberry. The analytical study was conducted by the method of determining the quality of fruit and berry products (kondratenko et al., 2008), in particular, using spectrophotometry methods, the content of biologically active substances was determined and expressed in mg/100 g FW. The amount of vitamin C was determined using titration with the solution of 2,6-dichlorophenolindophenol, the content of polyphenolic substances was determined using the Folin-Denis reagent, the content of flavonoids - by measuring the absorption of the complex of flavonoids by aluminum chloride in the ethanol medium, the content of anthocyanins - by the difference method in two buffer systems: with potassium chloride, & rcy;& Ncy; 1.0 (0.025 M) and sodium acetate, & rcy;& Ncy; 4.5 (0.4 & Mcy;). The statistical analysis of the study data was conducted in STATISTICA 13/1 (StatSoft, Inc., USA). Results. The weather indices of the period of growth and development of strawberry fruits had a considerable impact on the content of vitamin C, polyphenols, flavonoids, and anthocyanins in them. The content of vitamin C in the fruits of cultivar Herkules was the most stable and independent from weather conditions of the cultivation year, with a minimum of 50.4, a maximum of 57.6, and an average of 53.5 mg/100 g FW. The content of vitamin C in strawberry cultivars Florence and Vibrant was the most variable and dependent on the weather conditions of the cultivation year, the average content was 52.0 and 50.9 mg/100 g FW. The amount of vitamin C in strawberry fruits considerably depended on the average daily air temperatures; the correlation coefficients for the investigated group of cultivars were high, over 0.720, the increase in the average daily air temperatures promoted a decrease in the content of vitamin C. The amount of polyphenols in cultivars Herkules and Prezent had a significant correlation with the average daily air temperatures and the HTI for the period of their growth and development, but in the first case, the correlation was direct, and in the second one, - indirect. Among the investigated cultivars, the fruits of cultivars Veselka, Vibrant, and Florence were capable of accumulating the maximal amount of flavonoids, over 70 mg/100 g FW. The fruits of Prezent cultivar were capable of accumulating more than 20 mg/100 g FW of anthocyanins. The content of flavonoids in Prezent cultivar and that of anthocyanins in Veselka cultivar was homeostatic. Conclusions. It was determined that the content of biologically active substances in strawberry fruits was determined by genetic specificities of the cultivar and corrected by weather conditions for the period of their growth and development.
The aim of the study. To demonstrate the main regularities in the formation of moisturization regime and to determine the impact on the implementation of agropotential and restoration of fertility of chornozem under the effect of crop rotations, fertilization, and tillage in conditions of current climate changes in the central Forest-Steppe. Methods of studies. Field method (to select samples in the depth of chornozem in different study periods), laboratory method (for thermogravimetric determination of the productive moisture reserves during the study period), mathematical, statistical, and comparative evaluative methods (to substantiate the reliability of the obtained moisture reserves in soil and elaboration of statistical models of interactions). Results of studies. On average, in 75 years of observations, the average moisture reserves in a one-meter-deep layer were 61.6 mm in November, and 51.5 mm by the median, that tended to the lower typical value which demonstrated a decrease in autumn moisture reserves. The oscillation coefficients (& Kcy;& ocy;s) and & Kcy;var were 40% and 54.4%. The moisture reserves in April, on average and by the median value, were 156-155 mm at & Kcy;& ocy;s=37% and & Kcy;var=10.3%. The average accumulation of moisture in the one-meter-deep layer in the cold period of the year was 94 mm with the median value of 99 mm, which tended more to the higher typical value (L0.75) at & Kcy;& ocy;s=98% at & Kcy;var=25.3%. In July, the moisture reserves in the one-meter-deep layer were 50.1 mm and by the median value - 45.5 mm which demonstrated tending to the lower typical value (L0.25) and the intensification of drought conditions, when the index of moisturization (Im) was within 0.77-0.88 (weakly arid conditions), and at the maximal typical value of Im=1.13-1.22 (wet conditions) at & Kcy;os=65% and & Kcy;var= 18.6%. The average loss of moisture from soil in April-July was-105 mm, which corresponded to the median value and tended to the higher typical values of losses (-108...-122 mm) at & Kcy;os=33% and & Kcy;var=9.3%, which is a stabilized loss, tending to increase. Between the parameters of productive moisture content and climate indices, there were direct and indirect correlational relations of high correlation rate (R=+/- 0.61-0.95 +/- 0.02, R2=37-0.90), and the relations between Im and Dm increased to strong correlation (R=+/- 0.68-0.96 +/- 0.03, R2=0.46-0.65). It was found that in the 0-30 cm layer of podzolic chornozem, the average humus content after crop rotation with plowing was 2.34%, under surface tillage, the humus content increased by 0.15% (10 years of surface tillage), and in case of N & ocy;-till, the humus content increased by 0.07-0.08%. In the crop rotation on typical chornozem, the average reserves of & Scy;-& Scy;& Ocy;2 in the 0-30 cm layer of chornozem were 273 t/ha, and in the 0-20 cm soil layer - 182 t/ha. The average reserves of & Scy;-& Scy;& Ocy;2 under No-till were 19.8 t/ha higher compared to plowing, and the reserves of & Scy;-& Scy;& Ocy;2 in the 0-30 cm layer of soil started from 296 t/ha, which was 21-23 t/ha higher compared to plowing. Under surface tillage, the average reserves of & Scy;-& Scy;& Ocy;2 were 14 t/ha higher than under plowing. The reserves of sequestered & Scy;-& Scy;& Ocy;2 in the 0-30 cm layer corresponded to the interval of values of 285-300 t/ha and was higher than under plowing but tended to decrease compared to No-till. The reserves of & Scy;-& Scy;& Ocy;2 in the 0-20 cm chornozem layer were at the 5 t/ha) and exceeded the reserves of carbon oxide under plowing by 21 t/ha, which demonstrated the increase in the sequestered ability of the 0-20 cm layer under surface tillage or No-till. In modern climatic conditions, the agropotential of chornozem is maximally implemented in the central Forest-Steppe, and a type of crop rotation is a relevant factor in increasing agrocenosis performance. In a crop rotation with peas, saturated with cereals by 80%, including 40% of grain corn, the yield of cereals was 5.56-6.15 t/ha, which exceeded the crop rotation with perennial grasses and 20% corn 1.06-1.05 times, and the yield of cereals - 1.31-1.4 times. The content of digestible protein was 0.67 t/ha or 19.7% higher in the crop rotation with grasses. Long-term (10 years) application of 8-10 cm surface tillage in a cereal crop rotation ensured the yield of cereals at the plowing level, whereas in case of No-till for 5 years, the yield of cereals was considerably lower compared to plowing and surface tillage. Conclusions. The relation, determined between climate parameters and the response of a one-meter-deep chornozem layer in the form of normalized parameters of productive moisture reserves, allows for parameterizing the moisturization regime within the periodic washing water regime of the central Forest-Steppe of Ukraine. The analysis of weather and climate parameters in the complex with the formation of productive moisture reserves in one-meter-deep layer in spring and summer periods in 1947-2022 demonstrated that in the central part of the Left-Bank Forest-Steppe, water regime is formed which corresponds to the periodically washing type, but in terms of the moisturization regime (time-wise), there is a clear stable tendency towards the manifestation of features of non-washing water regime, which mostly indicates the aridization of soil conditions for the vegetation of crops in modern conditions of global climate change in the Forest-Steppe zone. Therefore, the typization of moisture change in the one-meter-deep chornozem layer in the seasonal cycle (moisturization regime) can be used as an indicator of response to synoptic changes within climatic factors in the formation of the periodic washing regime of the central Forest-Steppe of Ukraine in modern climatic conditions.
Aim. This study aimed to develop a specific and sensitive multiplex real-time reverse transcription PCR (RTqPCR) method for the detection and differentiation of rabbit hemorrhagic disease virus (RHDV) genotypes GI.1 and GI.2 circulating in Ukraine, using primers and probes based on the ORF2 gene, encoding the minor capsid protein VP10. Furthermore, to perform an initial validation of this PCR. Methods. The assay was designed to amplify a conserved 101-bp ORF2 sequence, encoding the minor capsid protein VP10, assuming that it would be a less variable region compared to that of the traditionally targeted ORF1 sequence encoding the major capsid protein VP60. Sequence alignment was performed using 38 full-genome sequences of RHDV isolates of various geographic origins present in GenBank. Specific primers to RHDV and two genotype-specific hydrolysis probes (FAM for genotype GI.1 and HEX for genotype GI.2) were designed and partially validated both in silico (BLAST) and in vitro. RNA was extracted from 6 tissue samples (contaminated with RHDV and negative control), and two virus reference strains using the IndiSpin Pathogen Kit, followed by RT-qPCR using a one-step protocol. This protocol and primer and probe sequences are detailed in the main text. Analytical sensitivity and specificity were assessed by a 10-fold RNA serial dilution (from 106 to 100 copies). For specificity testing, in addition to RHDV isolates, non-target organisms were included: Myxoma virus (strain B-82, IVM NAAS collection), Staphylococcus aureus subsp. aureus ATCC 25923, Pasteurella multocida subsp. multocida ATCC 12945, Escherichia coli ATCC 25922, and Streptococcus agalactiae ATCC 13813. The data were statistically analysed using R software and the detection limit (LOD) and sensitivity were determined. Results. After optimization, the developed RTqPCR assay demonstrated in a partial validation a high analytical sensitivity, detecting as few as 100 RNA copies/ reaction with consistent amplification across three replicates. The limit of detection (LOD) was established at 1.0x102 RNA copies for both genotypes in a dilution series of virus RNA. Standard curves based on Ct values versus log10 of RNA concentration yielded slopes of-3.44 (GI.1) and-3.38 (GI.2), corresponding to amplification efficiencies of 95.4% and 97.5%, respectively. The assay showed excellent linearity (R2 = 0.925 for GI.1, R2 = 0.881 for GI.2) and intra-assay variability (%CV) below 3.5% across all tested dilutions. No cross-reactivity was observed with rabbit myxomatosis virus and four bacterial pathogens. In a sample containing both genotypes, the assay successfully detected and differentiated both GI.1 and GI. 2 targets. Conclusions. In this study, we designed a multiplex real-time RT-qPCR assay targeting the VP10 gene (ORF2) for the detection and differentiation of rabbit hemorrhagic disease virus (RHDV) genotypes GI.1 and GI.2 circulating in Ukraine. The assay demonstrated promising analytical performance, with an estimated limit of detection of 100 RNA copies/mL and no observed cross-reactivity with selected non-target pathogens. These results provide a partial validation of the method, indicating its potential applicability for laboratory diagnosis and epidemiological investigations. However, several limitations must be acknowledged. The current validation was restricted to a limited number of samples and two genotypes, while GI.3, GI.4, GII.1, GII.2, and recombinant variants were not included. No internal amplification control was applied, and (inter-laboratory) comparison with the WOAH-recommended VP60-based RT-PCR has yet to be conducted. Future studies should complete the validation by expanding the range of tested genotypes, incorporating an internal control, and performing multi-center validation. Additional testing on portable diagnostic platforms and under field conditions will further determine the assay's suitability for routine veterinary practice. Pending such validation, the VP10-based multiplex RT-qPCR assay then may complement existing VP60-based diagnostics, or serve as a (preliminary) control of those classical diagnostics, when surveying RHDV in Ukraine.
The aim of the study was to evaluate the bioecological properties and reproductive potential of egg parasitoid Uscana senex (Grese, 1923) (Hymenoptera: Trichogrammatidae) with the purpose of using it within the programs of biological control over pea weevil Bruchus pisorum (Linnaeus, 1758) (Coleoptera: Chrysomelidae). Methods. The belonging of a parasite to a particular species was determined by morphological traits and a detailed analysis of the structure of the genitalia and the form of the imago club. Common entomological methods were used in laboratory and field conditions to study the impact of various ratios of parasite:host (from 1:1 to 1:45) on the reproductive characteristics of entomophages. The main viability indices of the laboratory and natural populations of the parasite were compared: motor activity, searching capacity, restoration rate, and the share of deformed insects. The impact of the periodic passage of a laboratory culture via the natural host on the quality of biomaterial was also evaluated. The statistical results were processed by the dispersion analysis (ANOVA), and the data were presented as x +/- SD. Results. The results demonstrated that the fecundity of female entomophages was increasing considerably with the increase in the host number; however, the efficiency of the parasite utilization decreased with the following generations. The highest biological efficiency in field conditions on the plots with Oplot pea cultivar was registered at the 1 : 5 ratio, where the parasitism rate was 39.8%, and the seed damage was minimal - 5.59%. Statistically significant differences between the native and laboratory populations of U. senex were found, and the native population demonstrated higher viability indices as compared to the laboratory population. The periodic passaging of the laboratory population via the natural host partially compensated for the viability loss and reduced reproductive potential. Conclusions. The obtained results demonstrated high efficiency of Uscana senex as a biological agent in the integrated systems of protecting pea from Bruchus pisorum. It was proven that maintaining the reproductive potential and viability of the parasite was ensured on condition of periodic contacts between the laboratory population and the natural host, and adherence to the optimal parasite:host ratio, which promotes the preservation of stable quality of the biomaterial.
Aim. To identify the criteria and select cyanobacterial species that may be used as biofertilizers for agricultural soils. Methods. Specific methods of synthesis, logical analysis, and correlation of scientific results, comparison, systematization, explanation, and deduction were used in the work. Field studies were conducted on chornozem (typical, common, and leached) under various crops (sunflower, wheat, barley, corn, rape, and pea) in the central and southern regions of the Republic of Moldova from 2010 to 2025. Results of studies. The criteria for selecting bacteria to be applied as biofertilizers on agricultural lands, used for the cultivation of crop plants, were established as follows: dominance, spreading, high tolerance to environmental factors (without intermediate accommodation stages), a short generation time, and potential for industrial cultivation. The cyanobacteria recommended as biofertilizers for agricultural soils were identified, and their taxonomic position, from genus to species, was presented. The most representative genera of cyanobacteria recommended as biofertilizers for various soil types are as follows: Microcystis, Nostoc, Anabaena, Cylindrospermum, Tolypothrix, Calothrix, Oscillatoria, Phormidium, Lyngbya, Schizothrix, and Microcoleus. Also, 65 species of cyanobacteria, representatives of the dominant and globally widespread genera, recommended for use as biofertilizers, were determined. The optimal dose of cyanobacterial fertilizers (ranging from 5 to 258 kg/ha) and the form of their administration to ensure the sustainability of the health of soils and crops were established. Conclusions. Cyanobacteria are promising biofertilizers with proven impact on soil health and crop productivity. The study defines the criteria for selecting effective species and highlights the recommended genera and species for various soil types. Their application in the living form may support the fertility and sustainability of agroecosystems.
Aim. To screen the Ukrainian collection of hop breeding material for the level of field resistance to Pseudoperonospora humuli at the early stages of the hop plant development and growth, and to identify valuable genotypes for further breeding. Methods. The research was conducted in years 2019-2022 in small experimental plots in a field where natural infection of P. humuli was present (Zhytomyr district, the Polissia zone, Ukraine). Sixtyfive newly created hop genotypes and four standard Ukrainian varieties, Alta, Sl & ocy;vyanka, Zahrava, and Ruslan (as controls), were evaluated for the field resistance against P. humuli on base of disease index using a 9-point scale during the initial growth phase of the plant at two stages: in stage I, emergence of sprouts and stage II, shoot growth. Results. For the first time a descriptive analysis of field resistance to P. humuli of hop plants in two stages of its development and growth was performed for 65 new genotypes from the Ukrainian collection of genetic hop resources and the most resistant ones were selected. The disease development was less intense in stage I than during stage II, which was possibly associated with increased humidity and temperature during the latter stage. No genotype showed immunity (full resistance) to the disease. The four standard varieties used showed moderate susceptibility. Eight genotypes (7667, 8156, 8195, 8367, 8382, 8388, 8604, and 8555) demonstrated high resistance during stage I, while a slightly different set of eight genotypes (7667, 7886, 8156, 8195, 8367, 8438, 8605, and 8555) showed high resistance in stage II. The differences observed across evaluation stages and genotypes were statistically significant (F=13.4; & rcy; < 0.001). Conclusions. Evaluation of the Ukrainian collection of hop breeding material for field resistance to Pseudoperonospora humuli during early stages of plant development and growth showed that 25% of the genotypes were classified as resistant, 39% as moderate resistant, 32% as moderate susceptible, and 4% as susceptible. Six genotypes (7667, 8156, 8195, 8367, 8382, and 8555) with the highest total field resistance to P. humuli were advised for inclusion into further breeding crossings and investigations for the presence of useful genetic resistance markers.
Aim. Single-nucleotide polymorphism analysis of maize inbreds with different rate of sensitivity to the head smut caused by Sporisorium reilianum, assessment of genetic relationships between them, and search for statistically significant associations between SNP markers and phenotypic manifestation of pathogen resistance. Methods. The single-nucleotide polymorphism analysis using BDI-III panel with 384 SNP markers, statistical methods. Results. The SNP analysis of seven maize inbreds with different sensitivity to S. reilianum infection was carried out. The key indicators of single-nucleotide polymorphism markers with BDI-III panel were assessed: the fraction of dimorphic markers was 85.8%, the average minor allele frequency was 0.2984, the fraction of dimorphic markers with the minor allele frequency >0.2 was 68.2%, the average polymorphism information content (PIC) was 0.2770. The main indicators of the maize inbreds were determined by SNP markers. It was established that the average homozygosity of the inbreds was 98.5%, the average gene diversity index was 0.1723, and the average genetic distance was 0.4975. It was found the allelic state of SNP markers of the BDI-III panel — BDI-III-270C and BDI-III-292A, which is characteristic of maize inbreds resistant to head smut, as well as BDI-III-270A and BDI-III-292G for maize inbreds sensitive to head smut. The dendrogram of genetic relationships for the studied maize inbreds was drawn. Conclusions. The presence of statistically significant associations between the allelic state of two SNP markers and the phenotypic manifestation of the maize resistance to head smut was established. The random nature of the location of maize inbreds sensitive to head smut on the dendrogram of genetic relationships was revealed. In the future, the identified single-nucleotide polymorphism markers, which were associated with head smut resistance, will be verified on a larger number of maize genotypes of different origin to confirm the possibility of their practical application.
The aim of the study was to analyze the parameters of the reproductive ability of Chinchilla doe rabbits with different genotypes by locus GH (mutation c.-78C>T). Methods. The genotypes of animals were determined by PCR-RFLP. The amplification was conducted in the programmed thermocycler Applied Biosystems MiniAmp (Thermo Scientific, the USA) using specific primers. The amplicons were cleaved with the endonuclease BstUI (Thermo Scientific, the USA), and the fragments were separated in 2% agarose gel supplemented with ethidium bromide. The following indices were used to evaluate the reproductive ability of doe rabbits: milk production (kg), number of litter kits, birth and weaning weight of litter (kg), and average daily gain of infant rabbits (g). The population and genetic indices were assessed using common methods. The impact of the genotype of GH locus on the parameters of the reproductive function of doe rabbits with different genotypes by locus GH was analyzed using ANOVA. Results. The investigated sampling of Chinchilla doe rabbits (34 animals) was found to have BstUI-polymorphism in position -78 of the start codon of exon I of gene GH (c.-78C>T). The analysis of associations demonstrated that the doe rabbits with the heterozygous genotype CT in locus GH had higher indices of milk production, number of litter kits and live weight of the litter, average daily gain, and live weight of one animal at the age of 35 days. Statistically significant differences were found between genotypes CT and TT in terms of milk production (p<0.001) and live birth and weaning weight of the litter (p<0.05). Conclusions. The results obtained demonstrated the promising perspectives of the use of BstUI-polymorphism of locus GH (c.-78C>T) as a genetic marker in the breeding programs, aimed at enhancing the fecundity of rabbits.
