Analysis of X-ray irradiation effects on the mortality values and hemolymph immune cell composition of Apis mellifera and its parasite, Varroa destructor.

Varroa destructor is one of the most destructive enemies of the honey bee, Apis mellifera all around the world. Several control methods are known to control V. destructor, but the efficacy of several alternative control methods remains unexplored. Irradiation can be one of these unknown solutions but before practical application, the effectiveness, and the physiological effects of ionizing radiation on the host and the parasite are waiting to be tested. Therefore, the objective of our study was to investigate the effects of different doses (15, 50, 100, and 150 Gy) of high-energy X-ray irradiation through mortality rates and hemocyte composition changes in A. mellifera workers and record the mortality rates of the parasite. The mortality rate was recorded during short-term (12, 24, and 48 h) and long-term periods (3, 6, 12, 18, and 24d). The sensitivity of the host and the parasite in case of the higher doses of radiation tested (50, 100, and 150 Gy) been demonstrated by total mortality of the host and 90 % of its parasite has been observed on the 18th day after the irradiation. V. destructor showed higher sensitivity (1.52-times higher than the adult honey bee workers) at the lowest dose (15 Gy). A. mellifera hemocytes were influenced significantly by radiation dosage and the elapsed time after treatment. The higher radiation doses increased plasmatocyte numbers in parallel with the decrease in prohemocyte numbers. On the contrary, the numbers of granulocytes and oencoytes increased in the treated samples, but the putative effects of the different dosages on the recorded number of these hemocyte types could not be statistically proven. In summary, based on the outcome of our study X-ray irradiation can be deemed an effective tool for controlling phoretic V. destructor. However, further research is needed to understand the physiological response of the affected organisms.

Biophoton emission-based approach of the effects of systemic insecticides on the survival of Eurydema ventralis Kolenati, 1846 (Hemiptera: Pentatomidae) and on the photosynthetic activity of oilseed rape.

The choice of effective crop protection technologies is a key factors in the economical production of oilseed rape. Insecticides belonging to the group of active substances butenolides and diamides are active substances available as seed treatments in oilseed rape and promising control tools in the crop protection technologies. Our laboratory experiment demonstrated that the experimental insecticides flupyradifurone and cyantraniliprole are both effective against Eurydema ventralis (Hemiptera: Pentatomidae) when used as a seed and in-crop treatments, but there is a fundamental difference in their insect mortality inducing effects. Flupyradifurone was found to have a total mortality 96 h after application based on basipetal translocation. In the case of cyantraniliprole, the insecticidal effect of the same treatment was 27% less. The experiment showed that the acropetal translocation of the tested active substances after seed treatment did not induce efficacy comparable to that of the basipetal translocation. The study of the biophoton emission of the plants demonstrated a verifiable correlation between the different application methods of the insecticides and the photon emission intensity per unit plant surface area. In conclusion, the systematic insecticides tested, in addition to having the expected insecticidal effect, interfere with plant life processes by enhancing photosynthetic activity.

New insight into the Delia platura Meigen caused alteration in nutrient content of soybean (Glycine max L. Merill).

Climate change has brought about an increasing level of seedcorn maggot (Delia platura Meigen, 1826) (Diptera: Anthomyiidae) damage in Hungary. In order to have a more accurate understanding of the effects of these plant injuries induced by the larvae of D. platura, the nutrient content of soybean (Glycine max L. Merill.) was studied. Our results show that the moisture, raw fat, raw fibre, and raw ash content of the batches damaged by D. platura were significantly less in comparison with that of the control samples. In response to the deleterious effect of the insect, the infected soybean plants showed forced ripening (P = 0.004) (P > 0.05). The difference of moisture content between damaged and control samples was 2.30% on average. The fact of nutritional value loss was also reflected by the alteration of sugar mobilisation. As the result of this experiment the sucrose breakdown to glucose and fructose during the germination was significantly slower in the damaged seeds than that of the control ones. Overall, this late and surprising damage caused by D. platura led to the forced ripening of the affected soybean plants and a significant change in their nutritional values. Based on the herein reported results, it is presumable that in cases when the current climatic extremities, which are envisaged to occur more frequently in the future, and effects of agricultural practices will be coincided in the future a qualitative change of the produced soybean batches can be expected through the damage caused by this fly species.

Detection of damage caused by Nezara viridula on soybean using novel imaging approaches based on computed tomography and image color analysis.

Soybean (Glycine max L.) is an important leguminous plant, in which pests trigger significant damage every year. Important members of this community are insects with piercing-sucking mouthpart, especially the southern green stinkbug, Nezara viridula L.. This insect with its extraoral digestion causes visible alterations (morphological and color changes) in the seeds. We aimed to obtain precise information about the extent and nature of damage in soybeans caused by N. viridula using nondestructive imaging methods. Two infestation conditions were applied: one with controlled numbers of pests (six insects/15 pods) and another with naturally occurring pests (samples collected from the apical part of the plant and samples from whole plants). An intact control group was also included, resulting in four treatment groups. Seed samples were analyzed by computed tomography (CT) and image color analysis under laboratory conditions. According to our CT findings, the damage caused by N. viridula changed the radiodensity, volume, and shape (Solidity) of the soybean seeds during the pod-filling and maturing period. Radiodensity was significantly reduced in all three damaged categories compared to the intact sample; the mean radiodensity reduction range was 49-412 HU. The seed volume also decreased significantly (25%-80% decrease), with a threefold reduction for samples exposed to regulated damage compared to natural ones. The samples exposed to natural damage showed significant but minor reduction in solidity, while samples exposed to regulated damage showed a prominent decrease (~12%). Image color analysis showed that the damaged samples were well distinguishable, and the differences were statistically verifiable. The achieved data derived from our external and internal imaging approaches contribute to a better understanding of the internal chemical processes, and CT analysis helps to understand the alteration trends of the hidden structure of seeds caused by a pest. Our results can contribute to the development of a practically applicable system based on image analysis, which can identify lots damaged by insects.

Identification and investigation of barley powdery mildew (Blumeria graminis f. sp. tritici) infection in winter wheat with conventional stress reactions and non-invasive biophoton emission parameters.

The objective of this study was to characterize the effects of barley powdery mildew infection on wheat via the evolution and dynamics of chloroplasts and oxidative processes based on in vivo measurements of ultra-weak photon emission, parallel measurement of chlorophyll and ascorbic acid content, and molecular identification of the pathogen. The results showed the temporal dynamics of the evolution of ultra-weak photon emission signals that were evidently different for healthy and powdery mildew-infested wheat leaves. In the dark, the ceasing of delayed fluorescence signal made it possible to visualize the ultra-weak luminescence signal as well. Both delayed fluorescence and ultra-weak luminescence signals were characteristic of stress symptoms induced by powdery mildew that was further strengthened by the changes of chlorophyll and ascorbic acid content as typical stress analytical parameters. The presented data and parameterization enabled the identification of stress induction due to powdery mildew infestation in wheat, which should be investigated in detail in the future for fine-tuning our measurements, even by using other species and increasing the length of the measurement in order to increase its specificity. The changes in R2 values are suitable for monitoring the changes of plant stress response. The measurement of fluorescence and luminescence leads to a greater comprehension of the underlying photon emission-related processes, both in general and in the case of powdery mildew infestation.

Tracking Varroa Parasitism Using Handheld Infrared Cameras: Is Eusocial Fever the Key?

The Varroa destructor is the most significant bee parasite and the greatest threat to bee health all around the world. Due to its hidden lifestyle, detection within the brood cell is only possible through invasive techniques. Enhancing detection methods is essential for advancing research on population dynamics, spread, selection efforts, and control methodologies against the mite. In our study, we employed infrared imaging to measure the thermal differences in parasite and intact Apis mellifera worker broods. Experiments were conducted over two years at the MATE Kaposvár Campus in Hungary involving five beehives in 2022 and five beehives in 2023. A FLIR E5-XT WIFI handheld infrared camera was used to create a heat map of capped brood frames. Our results indicate that the resolution of these cameras is sufficient to provide detailed IR images of a bee colony, making them suitable to detect temperature differences in intact and Varroa parasitized capped brood cells. Mite parasitism causes a time-dependent and sustained temperature increase in developing bee pupae, observable regardless of mite number. Our work demonstrates two different heating patterns: hotspot heating and heating cells that are responsible for the elevated temperature of the Varroa-infested cells as a social fever response by the worker bees. Based on our results, future research combined with AI-based image evaluation software could offer beekeepers and researchers practical and valuable tools for high-throughput, non-invasive Varroa detection in the field.

Impact analysis of different applications of cyantraniliprole on control of horse chestnut leaf miner (Cameraria ohridella) larvae supported by biophoton emission.

Cameraria ohridella is one of the most invasive pests of horse chestnut. Cyantraniliprole is one of the most perspectively active insecticides, which can transport within the plant in several ways, and its efficacy against this pest has not yet been tested. All three modes of application were effective against the target pest, but there was a difference in the time of action between them. However, no demonstrable difference in the speed of action was detected between the doses used. A more intense rate of acropetal translocation was confirmed compared to basipetal translocation. A trend-like effect between the applied concentration of cyantraniliprole and the photon emission intensity per unit area of plant tissue was observed in the translaminar and acropetal treatment settings. In both cases, a clear increase in photon emission was observed, indicating metabolic upregulation. Therefore, we can conclude that biophoton emission measurements can be utilized to conduct efficient pesticide translocation investigations.

Physiological Responses Manifested by Some Conventional Stress Parameters and Biophoton Emission in Winter Wheat as a Consequence of Cereal Leaf Beetle Infestation.

Oulema melanopus L. (Coleoptera: Chrysomelidae) is one of the most serious pests of winter wheat that causes peeling of the epidermis and tissue loss. The complex mapping of the physiological responses triggered by O. melanopus as a biotic stressor in winter wheat has not been fully explored with the help of non-invasive imaging and analytical assays, yet. The aim of the present work was to study the effect of O. melanopus on the physiological processes of winter wheat, especially on the extent of lipid peroxidation and antioxidant activity derived from tissue destruction, as well as photosynthetic ability. The results of the measurements enabled the identification of the antioxidant and lipid-oxidation-related physiological reactions, and they were reflected in the dynamics of non-invasive biophoton emissions. Our non-invasive approach pointed out that in the case of O. melanopus infestation the damage is manifested in tissue loss and the systemic signaling of the biotic stress may have reached other plant parts as well, which was confirmed by the results of antioxidant capacity measurements. These results indicate that the plant reacts to the biotic stress at a whole organizational level. We identified that the antioxidant and lipid-oxidation-related physiological reactions were reflected in the dynamics of two aspects of biophoton emission: delayed fluorescence and ultra-weak bioluminescence. Our research further supported that the non-invasive approach to stress assessment may complete and detail the traditional stress indicators.

Analysis of the Destructive Effect of the Halyomorpha halys Saliva on Tomato by Computer Tomographical Imaging and Antioxidant Capacity Measurement.

Qualitative and quantitative parameters of tomatoes are impaired by Halyomorpha halys Stål (Hemiptera: Pentatomidae), which cause severe economic losses worldwide. Our aims were to assess H. halys-induced tissue damage in tomatoes via computer tomography and to confirm the results of imaging obtained by analytical methods. Our examination confirmed the intensification of the change in the inner structure of damaged tomatoes as a function of time. The tendency of this destruction triggered by bug saliva grew exponentially from the exocarp layer to the inner placenta. The destruction of the plant tissue was aggravated by an increase in the number of bugs, as it was unequivocally evinced by the shell thickness assays. The results of the assessment of the antioxidant capacity of tomato mesocarp showed a distinct decrease in the antioxidant capacity of the samples obtained from H. halys-infested tomatoes. This indicates that the ferric-reducing antioxidant power value was related to the degradation processes of the mesocarp tissue in tomato fruit caused by the watery saliva released by H. halys. The presented experimental method can be suitable for the qualitative control of the vegetable items intended for trade, which can help for the isolation of tomatoes damaged by bugs immediately after harvest.

Non-Invasive Evaluation of Different Soil Tillage and Seed Treatment Effects on the Microbial Originating Physiological Reactions of Developing Juvenile Maize.

The successful production of maize is fundamentally determined by a good choice of tillage type. Options include conventional tillage based on soil rotation, as well as a more recent conservation approach. Our aims were to determine the stress physiological effects of the plant remains left behind by different tillage procedures on the juvenile maize plants, combined with the effects of fungicide treatment on the seeds. These effects were followed and investigated by means of biophoton emission measurement, an in vivo and non-invasive imaging technique, along with chlorophyll content estimation, as well as microbial- and polymerase chain reaction-based identification of fungi presence. Our results confirmed the response reactions of maize triggered by a soil covering plant remains on the initial development and physiological involvement of maize. The positive effects of seed treatment on initial development are manifested only at the final stage of the experiment. The fungal microbiological analysis confirmed the dominant presence of necrotrophic parasites on plant residues, the stress-inducing properties of which were possible to monitor by biophoton emission. Furthermore, the presence of Fusarium spp. was confirmed by PCR analysis from samples treated with plant residues.

Effects of Different Infra-Red Irradiations on the Survival of Granary Weevil Sitophilus granarius: Bioefficacy and Sustainability.

Sitophilus granarius (L.) is an important pest of stored grain worldwide. In recent years, sustainable methods against it have received attention as grain stock protective means. Our aim was to obtain information about the efficacy of infrared irradiation (IR) against S. granarius in laboratory conditions. The change in adult-activity and median lethal dose (LD50) triggered by IR in S. granarius was examined. The insecticidal efficacy in the infested grains was also analyzed at 12, 24, 48, and 72h following exposure to IR (250W), and the progeny-production was assessed 45 days upon the treatment. Based on our findings, total mortality ensued in a grain stock of 50 g at 412 s and a for 100 g grain at 256 s. A significant increase in S. granarius mortality could be observed in the higher grain weight regime, which can be accounted for by the higher heat-absorbance of objects with higher weight. The activity of pests immediately after the beginning of IR increased and subsequently became moderated. The observation of activity-peak brought about by irradiation contribute to the optimization of chemical intervention. This treatment could provide an effective and sustainable technique in integrated pest management.

Study of Morphological Features in Pre-Imaginal Honey Bee Impaired by Varroa destructor by Means of Computer Tomography.

The honey bee (Apis mellifera L. 1778) is an essential element in maintaining the diversity of the biosphere and food production. One of its most important parasites is Varroa destructor, Anderson and Trueman, 2000, which plays a role in the vectoring of deformed wing virus (DWV) in honey bee colonies. Our aim was to measure the potential morphometric changes in the pre-imaginal stage of A. mellifera caused by varroosis by means of computed tomography, hence supplying evidence for the presumable role that V. destructor plays as a virus vector. Based on our results, the developmental disorders in honey bees that ensued during the pre-imaginal stages were evident. The total-body length and abdomen length of parasitized specimens were shorter than those of their intact companions. In addition, the calculated quotients of the total-body/abdomen, head/thorax, and head/abdomen in parasitized samples were significantly altered upon infestation. In our view, these phenotypical disorders can also be traced to viral infection mediated by parasitism, which was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Capitalizing on a non-destructive method, our study reveals the deformation of the honey bee due to mite parasitism and the intermediary role this pest plays in viral infection, inside the brood cell.

A Non-Invasive Approach in the Assessment of Stress Phenomena and Impairment Values in Pea Seeds Caused by Pea Weevil.

Pea (Pisum sativum L.) is an important leguminous plant worldwide, in which pests trigger significant damage every year. One of the most important pest is pea weevil (Bruchus pisorum, L) which causes covert damage in crops. In the present study, our aim was to obtain precise information pertaining to the extent and the nature of damage in pea caused by B. pisorum by means of non-invasive imaging methods. The infested pea samples were analysed by an infrared thermometer and a bioluminescence plant imaging system as well as a computer tomograph under laboratory conditions. The calculated weight of organic matter destroyed by the developing larvae was 36.46%. The changing of RGB (red, blue, green) codes obtained through thermal imaging and the CPS (counts per second) values originating from bioluminescence imaging in infested samples were statistically verifiable. According to our CT assay, the damage caused by B. pisorum changed the tissue density, volume and shape of the pea seeds by the end of the development of the pest. The results of thermal and bioluminescence imaging contribute to a better understanding of the internal chemical processes and the CT analysis helps to understand the alteration trends of the inner structure of seeds caused by this pest.