Entomological Methods Research Articles

Environmental predictors impact microbial-based postmortem interval (PMI) estimation models within human decomposition soils.

Microbial succession has been suggested to supplement established postmortem interval (PMI) estimation methods for human remains. Due to limitations of entomological and morphological PMI methods, microbes are an intriguing target for forensic applications as they are present at all stages of decomposition. Previous machine learning models from soil necrobiome data have produced PMI error rates from two and a half to six days; however, these models are built solely on amplicon sequencing of biomarkers (e.g., 16S, 18S rRNA genes) and do not consider environmental factors that influence the presence and abundance of microbial decomposers. This study builds upon current research by evaluating the inclusion of environmental data on microbial-based PMI estimates from decomposition soil samples. Random forest regression models were built to predict PMI using relative taxon abundances obtained from different biological markers (bacterial 16S, fungal ITS, 16S-ITS combined) and taxonomic levels (phylum, class, order, OTU), both with and without environmental predictors (ambient temperature, soil pH, soil conductivity, and enzyme activities) from 19 deceased human individuals that decomposed on the soil surface (Tennessee, USA). Model performance was evaluated by calculating the mean absolute error (MAE). MAE ranged from 804 to 997 accumulated degree hours (ADH) across all models. 16S models outperformed ITS models (p = 0.006), while combining 16S and ITS did not improve upon 16S models alone (p = 0.47). Inclusion of environmental data in PMI prediction models had varied effects on MAE depending on the biological marker and taxonomic level conserved. Specifically, inclusion of the measured environmental features reduced MAE for all ITS models, but improved 16S models at higher taxonomic levels (phylum and class). Overall, we demonstrated some level of predictability in soil microbial succession during human decomposition, however error rates were high when considering a moderate population of donors.

An easier life to come for mosquito researchers: field-testing across Italy supports VECTRACK system for automatic counting, identification and absolute density estimation of Aedes albopictus and Culex pipiens adults

BackgroundDisease-vector mosquito monitoring is an essential prerequisite to optimize control interventions and evidence-based risk predictions. However, conventional entomological monitoring methods are labor- and time-consuming and do not allow high temporal/spatial resolution. In 2022, a novel system coupling an optical sensor with machine learning technologies (VECTRACK) proved effective in counting and identifying Aedes albopictus and Culex pipiens adult females and males. Here, we carried out the first extensive field evaluation of the VECTRACK system to assess: (i) whether the catching capacity of a commercial BG-Mosquitaire trap (BGM) for adult mosquito equipped with VECTRACK (BGM + VECT) was affected by the sensor; (ii) the accuracy of the VECTRACK algorithm in correctly classifying the target mosquito species genus and sex; (iii) Ae. albopictus capture rate of BGM with or without VECTRACK.MethodsThe same experimental design was implemented in four areas in northern (Bergamo and Padua districts), central (Rome) and southern (Procida Island, Naples) Italy. In each area, three types of traps—one BGM, one BGM + VECT and the combination of four sticky traps (STs)—were rotated each 48 h in three different sites. Each sampling scheme was replicated three times/area. Collected mosquitoes were counted and identified by both the VECTRACK algorithm and operator-mediated morphological examination. The performance of the VECTRACK system was assessed by generalized linear mixed and linear regression models. Aedes albopictus capture rates of BGMs were calculated based on the known capture rate of ST.ResultsA total of 3829 mosquitoes (90.2% Ae. albopictus) were captured in 18 collection-days/trap/site. BGM and BGM + VECT showed a similar performance in collecting target mosquitoes. Results show high correlation between visual and automatic identification methods (Spearman Ae. albopictus: females = 0.97; males = 0.89; P < 0.0001) and low count errors. Moreover, the results allowed quantifying the heterogeneous effectiveness associated with different trap types in collecting Ae. albopictus and predicting estimates of its absolute density.ConclusionsObtained results strongly support the VECTRACK system as a powerful tool for mosquito monitoring and research, and its applicability over a range of ecological conditions, accounting for its high potential for continuous monitoring with minimal human effort.Graphical

МЕТАГЕНОМНЫЙ АНАЛИЗ ВИРОМА КОМАРОВ AEDES (STEGOMYIA) ALBOPICTUS SKUS. ЧЕРНОМОРСКОГО ПОБЕРЕЖЬЯ КАВКАЗА

The aim of the study was to identify viruses contained in female Ae. albopictus mosquitoes using de novo metagenomic sequencing. Mosquitoes were collected using standard entomological methods in August 2019 in the Kuban in Tuapse, Adler, on the territory of the Adler Monkey Nursery and in the city of Gagra, Abkhazia. Adults were identified by morphological characteristics. A total of 1,336 mosquitoes were caught. We formed four pools at the collection site, with 300 to 400 specimens in each. The virus was considered detected if a specific nucleotide sequence of at least 1000 bases was detected in the mosquito sample. A total of 8 insect viruses were identified. Four viruses are classified and belong to the families Rhabdoviridae, Orthomyxoviridae, Chuviridae. The families of the other four viruses have not yet been determined, but two viruses are classified in the order Bunyavirales. No viruses pathogenic for humans were detected.

The effectiveness of plant protection products against Lobesia botrana Den. U Schift. in Zakarpattya

Goal. To establish the effectiveness of chemical protection of vineyards against pests, taking into account the peculiarities of their biology, distribution, harmfulness, seasonal dynamics of flight in Zakarpattya Ukraine. Methods. The research was conducted in accordance with the generally accepted methods in entomology and viticulture in 2021—2022 on the basis of Chateau Chizay LLC. The dynamics of the pest’s flight were studied in the field using the pheromone monitoring method using «Atrakon» pheromone traps and synthetic sex pheromones from the company «Csalomon» (Hungary). The research was conducted on Muscat, Chersegui, Sovignon Blanc, Pinot Noir, Cabernet Sovignon, Merlot, Rhine Riesling, and Pino Blan varieties. In 2022, in the vineyards of Chateau Chizay LLC, Zakarpattia Oblast, chemical plant protection agents were tested against grape leaf curl: Karate Zeon 050 CS, SK (liambda-tsyhalotryn, 50 h/l), 0.65 l/ha, Korahen 20, KS (khlorantraniliprol, 200 h/l), 0.15 l/ ha, Mavrik, EV (tau-fliuvalinat, 240 h/l), 0.2 l/ha, Voliam Fleksi 300 SC, KS (tiametoksam, 200 h/l + khlorantraniliprol, 100 h/l), 0.3 l/ha according to generally accepted methods. Results. Prolonged spring cooling in 2022 delayed the start of the flight of the first generation of cluster leafhopper, which began only in the first days of May. The second generation of this pest has been actively developing since the end of June. Grape damage on different varieties ranged from 2.1% on Muscat to 8.0% on Pino Blan. In the control (not treated with insecticides) plots, this indicator averaged 96.1%. The yield in the farm by variety averaged about 8 t/ha and ranged from 7.1 t/ha for the Muscat variety to 9.0 t/ha for the Merlot variety. In the control plot, the yield was low, only 1.4 t/ha. The effectiveness of protective measures on all grape varieties reached 90.5%. Conclusion. It was established that the protracted spring cooling caused the late start of flight of the grape leafhopper, which made adjustments to the calendar of protective measures, especially chemical ones. At the same time, thanks to pheromone monitoring, it was possible to establish the exact date of the start of Lobesia botrana’s flight and specify the timing of the application of chemical insecticides at the most favorable time, which made it possible to achieve an efficiency of more than 90%. Use of plant protection products against Lobesia botrana Den. et Schiff. in vineyards, LLC «Chateau Chizay», Zakarpattia Oblast contributed to the reduction of grape damage up to 94% and the preservation of the harvest up to 6.5 t/ha.

Species composition of phytophages on ornamental plants

Goal. Monitoring the phytosanitary status and determining the species composition of phytophages on decorative coniferous and deciduous plants. Methods. Field — determination of the species composition of pests and the dynamics of the number of main species; laboratory — determination of pests selected on experimental plots. Research was conducted in accordance with generally accepted methods in entomology and plant protection. Results. In the industrial plant nursery of open soil «Megaplant» located in the Kyiv region, Fastiv district, village The monitoring of the phytosanitary state of ornamental plant plantations was carried out in Trilys. The natural saturation of harmful organisms and the level of their harmfulness were studied. Conclusions. It has been established that plants in industrial nurseries in open ground are damaged by a large number of insects and mites. The most dangerous and widespread on conifer ornamental plants were 8 pests from the following families coccids (Coccidae), bark beetles (Scolytidae), adelgids (Adelgidae), snout beetles (Curculionidae) and spider mites (Tetranychidae); 9 most harmful species from the following families were noted on decorative leafy plants: crambid Snout Moths (Crambidae), mealybugs (Pseudococcidae) and аrmored scales or diaspidids (Diaspididae), eriophyidae mites (Eriophyidae), aphids (Aphididae), snout beetles (Curculionidae), scarab beetles (Scarabaeidae).

New and interesting records of rare caddisflies (Trichoptera, Insecta) from Slovakia with comments on their ecology and conservation status

The distribution and diversity of insects are crucial conservation issues. The order Trichoptera is comprised of numerous endangered species, but the rarity of some makes it challenging to determine their conservation priority. In recent decades, data based on caddisfly larvae have become more frequent in limnological research in Slovakia. Our research, focusing mainly on adult caddisflies, has led to more reliable and accurate identification of many species that cannot be identified with certainty in the larval stage.We combined several entomological methods, including sweeping, Malaise traps, and UV light attraction, to increase the likelihood of catching rare species. The text lists and discusses the discoveries of caddisfly species that are particularly important for faunistics or biodiversity in Slovakia, as well as in the Central European context. Enoicyla reichenbachii (Kolenati, 1848), Limnephilus incisus Curtis, 1834, Ptilocolepus granulatus (Pictet, 1834) and Oecetis testacea (Curtis, 1834) were recorded for the first time in Slovakia. The rare species Hagenella clathrata (Kolenati, 1848) and Adicella balcanica Botosaneanu and Novák, 1965, previously found in Slovakia, were also recorded. Additionally, a new locality of Adicella syriaca Ulmer, 1907 indicates a further northward shift of this Mediterranean species. We further discussed the faunistically important records of Beraeamyia hrabei Mayer, 1937, Allotrichia pallicornis (Eaton, 1873), Ceraclea riparia (Albarda, 1874), Grammotaulius nitidus (Müller, 1764) and Limnephilus fuscicornis Rambur, 1842 from Slovakia. The conservation status of each species is proposed and discussed on the basis of our faunistic records, in accordance with the forthcoming update of the Slovak Red Book.

Role of entomological evidence in criminal investigations

Domestic and foreign literary sources on the use of necrotic fauna in forensics are analyzed. The purpose of this article is to emphasize the importance of necrotic fauna research for practical forensics and highlight the influence of necrotic entomophiles on the time of death of a living object (depending on its species and environmental conditions) which biological and general scientific methods are applied for. A comparative analysis of the activity of necrotic fauna representatives is given. The need to use necrofauna in the systematic work of forensic biologists has been updated. The main application of forensic entomology as a branch of applied entomology is the study of the faunal complex of necrobiont arthropods on a discovered corpse to meet the needs of the investigation and the court. Entomological knowledge makes it possible to successfully perform classification, identification and diagnostic tasks that appear in forensic expert practice. Since the processes of corpse decay and the types of necrotic insects on it differ depending on the type of corpse and the type of biogeocenosis where this corpse was located, each of these features affects further forensic investigations. Obtained entomological data are a reliable source of information for finding out objective truth during pre-trial investigation and subsequent legal proceedings. At the same time, determining the time of death by cadaveric fauna can have only auxiliary value, since the appearance and development of insects is very variable. Prospects for further research are formulated, including improvement of entomological research methods (in particular, in different environmental conditions), the need for development of forensic biology as an integral component of forensic science is proven.

Hyperspectral lidar for monitoring high-resolution activity patterns of African stingless bee species

BackgroundStingless bees are vital pollinators and honey producers in the tropics. Research on stingless bees is generally underrepresented compared to the western honeybees, and while stingless bee studies from some regions are reported, there is a particular lack of reports on the species endemic to Sub-Saharan Africa. Since conventional entomological methods such as mark-recapture and radar harmonic tags suffer from limited observation counts and amount to a significant payload, fluorescent powder tagging offers a promising alternative to understanding their behavior. We deploy a hyperspectral fluorescence lidar monitors a 25-mm-wide transect in front of the hives.ResultsDuring a 1 day study at the International Stingless Bee Center, near Kakum National Park, Ghana, 17,862 insects were observed with the lidar, of which 7520 were tagged with fluorescent dyes. Approximately half of the bees from the selected hives were successfully tagged, with an estimated misclassification of 1%. According to our limited data, the observed species, Meliponula bocandei and the Dactylurina staudingeri exhibited different activity patterns. D. staudingeri displayed a half-hour longer active day, with clear crepuscular activity peaks. In contrast, M. bocandei activity was diurnal, with less pronounced crepuscular peaks.ConclusionsWe demonstrate how hyperspectral fluorescence lidar can monitor powder-tagged insects throughout the day. The monitored species revealed distinct activity patterns over the day. Our findings highlight the potential of this technology as a valuable tool for understanding insect behavior and environmental preferences of species, in situ, which could potentially give clues of response to climate changes of these critical species.

Moths are less attracted to light traps than they used to be

As evidence of global insect declines continues to mount, insect conservationists are becoming increasingly interested in modeling the demographic history of at-risk species from long-term survey data. However, certain entomological survey methods may be susceptible to temporal biases that will complicate these efforts. Entomological light traps, in particular, may catch fewer insects today than they once did due solely to increases in anthropogenic light pollution. Here we investigate this possibility by comparing the demographic histories of corn earworm moths (Helicoverpa zea) estimated from pairs of blacklight and pheromone traps monitored at the same farms. We find a stark decline in blacklight trap efficacy over 25 years of monitoring in Delaware, USA, mirrored over 10 years of monitoring in New Jersey, USA. While the precise causes of this decline remain a subject for discussion, the practical consequences are clear: insect conservationists cannot fully rely on long-term trends from entomological light traps.

Entomocomplex of Radish in Open-field Cultivation in the Central Part of the Right-Bank Forest-Steppe of Ukraine

Goal. To specify the species composition of insect pests affecting radish (Raphanus sativus (L.) convar. radicula (Pers) Sazon.) in open-field cultivation conditions in the central part of the Right-Bank Forest-Steppe of Ukraine.&#x0D; Methods. The study was conducted following commonly accepted methods in entomology and vegetable growing. From 2008 to 2022, we investigated the species diversity of insects, dominant species, their abundance, and the structure of the harmful entomocomplex in radish crops grown in open-field conditions on household plots and farms in the Cherkasy region.&#x0D; Results. In the entomological complex of radish, 59 species of phytophagous insects from 20 families and 8 orders were identified: Coleoptera — 29%, Lepidoptera — 27%, Homoptera — 15%, Diptera — 10%, Orthoptera — 9%, Thysanoptera — 5%, Hemiptera — 3%, Hymenoptera — 2%. Sixteen dominant species of phytophagous insects were found in radish crops, including diamondback moth (Plutella maculipennis Curt.), crucifer flea beetles (Phyllotreta cruciferae Goeze), striped flea beetle (Phyllotreta undulata Kutsch.), cabbage white butterfly (Pieris brassicae L.), turnip moth (Agrotis segetum Denis&amp;Schiff.) and owlet moth (Lacanobia oleracea L.), crucifer shield bug (Eurydema ventralis Kol.), cabbage aphid (Brevicoryne brassicae L.), spring root maggot (Delia brassicae Bouche) and summer root maggot (Delia floralis Fallen), seedcorn maggot (Delia platura Mg.), sawfly (Athalia rosae L.), stem weevil (Ceutorrhynchus quadridens Panz.), tobacco thrips (Thrips tabaci Lindeman), turnip sawfly (Entomoscelis adonidis Pallas), and crucifer webworm (Evergestis extimalis Scop.). Through multi-year studies, the periods of insect damage corresponding to radish developmental stages were determined. The most critical damage to radish plants by harmful insects occurred during the initial stages of development (ВВСН 0—9) and from emergence to the growth and formation of the root crop (BBCH 42—48).&#x0D; Conclusions. It was found that among the identified phytophagous insects in radish crops, the most economically significant damage was caused by the cabbage aphid, crucifer flea beetles, striped flea beetle, spring cabbage fly, and seedcorn maggot. On average over the research period (2008—2022), the largest areas of radish crops were infested by the diamondback moth, crucifer flea beetles, and striped flea beetle, cabbage white butterfly, winter and owlet moths, cabbage shield bug, and cabbage aphid, ranging from 5% to 80%, reaching a maximum in some years up to 60—100% of the area.

Monitoring of the species composition and number of insects and mites on the apple trees in the Right Bank Forest Steppe of Ukraine

Goal. To monitor the species composition and abundance of insects and mites on apple trees in the Right-Bank Forest-Steppe of Ukraine.&#x0D; Methods. Field, laboratory, microscopic, mathematical, statistical and calculation methods. The research was carried out according to generally accepted methods in entomology and plant protection, starting in early spring (late April — early May before harvest).&#x0D; Results. Monitoring of the number of insect pests and mites on apple trees in three types of plantations was carried out: a young orchard on stunted rootstocks with an intensive protection system; an old orchard on tall rootstocks with a protection system; an old orchard without treatments (long-term control).&#x0D; Conclusions. Monitoring of species composition and dynamics of pests number makes it possible to study and substantiate the patterns in the structure of insect and mite populations, to predict their harmfulness and to improve the protection system of apple trees or its individual components.

Plant viruses in the system of seed potato production

Solanum tuberosum L. is susceptible to 40 different virus species and 2 viroids. To prevent plant viruses from spreading in field conditions, it is necessary to have reliable data on the species composition of plant reservoirs of viral infection, the total activity of insect vectors, and possible ways of virus transmission in a particular territory of seed material production. Attention should be paid to the factors that facilitate and hinder the disease development in crops and to disease symptoms in different potato varieties. Manifestations of viral infections were monitored on every plant from the sample at the stages of initial growth, bud formation, and flowering and before the removal of potato haulms. Insects were collected using standard entomological method. The total RNA was isolated employing commercial kits for the extraction of nucleic acids from plant material “PhytoSorb” (Syntol Llc) and the benchtop automated extraction instrument KingFisher Flex (ThermoScientific) with magnetic particles. Plant viral infection was observed to accumulate if potato planting material was not renewed. The tested potato plants contained mixed viral infection, which consisted of viruses from mosaic group: PVY, PVX, PVM, PVS PVA, as well as PSTVd and PLRV. Without the renewal of seed potatoes, the concentration of plant viruses in an agroecosystem rises and causes secondary infections in potato plants. The research identified the main insect-vectors in the agroecosystem of potato fields: insects from genera Cicadella, Henosepilachna vigintioctomaculata, Dolycoris baccarum, Mythimna separata, Lygus pratensis, and Rhopalosiphum padi. Many wild weeds serve as fodder plants for insect vectors facilitating the accumulation of plant viruses in agroecosystems. It was established that perennial weeds were the main plant reservoirs of dangerous viral infections, e.g. Sonchus arvensis and Taraxacum officinale. We determined that Trifolium pratense typus L., Chenopodium album L., Plantago major L., Barbarea vulgaris W.T. Aiton, and Ambrosia artemisiifolia L. were the reservoirs of PVY. All these factors can lead to an epiphytotic situation.

Monitoring of the phytosanitary condition of Larix decidua Mill. plants in the Modryna tract of the Zviahel Forestry branch of the State Enterprise Forests of Ukraine

Phytosanitary monitoring of forest stands is currently a relevant and important area of scientific research that ensures observation, assessment, and forecasting of the dynamics (changes) of the quantitative and qualitative (including sanitary) state of forests and underlies the development of an effective system of tree protection. The purpose of this study was to identify the species composition and distribution of dominant phytopathogen species in the Modryna tract of the Zvyagelske Forestry branch and to identify the primary factors of Larix decidua Mill. decline. This study employed phytopathological, entomological, forest inventory, and comparative methods. The study identified and registered typical symptoms of damage and the consequences of the harmful effects of pests of L. decidua trees in the Modryna tract, leading to a general weakening of their sanitary condition. Symptoms of larch needle cast (meria needle blight) and browning were recorded on the needles; on the trunks – typical manifestations of cankers and fruiting bodies of a unique basidiomycete – Lariciphomes officinalis. Apart from infectious pathologies, sporadic wind and storm damaged larch trees, as well as other negative effects of wind exposure, were recorded in the study areas. The species composition of the pests of the surveyed larch trees included the following insect species: Cephalcia lariciphila, Sacchiphantes viridis Ratz., Coleophora laricella, Zeiraphera diniana Gn., Lasiomma laricicola Karl. It was generalised that the main threat to the centuries-old larch trees is the complex effect of anthropogenic, abiotic, and biotic (including parasitic) environmental factors that constantly and to varying degrees affect the sanitary and physiological state of ancient larch trees. The findings of this study provide information for predicting the risks of spreading pathogens and pests, which allows managing these risks and preventing their further spread

Updated Checklist of Butterfly Diversity (Lepidoptera: Papilionoidea) of Qatar

Butterflies (Lepidoptera: Papilionoidea) with their vibrant colours and ecological significance serve as crucial environmental indicators. This study documents the butterfly species inhabiting the challenging hot and arid climate of Qatar. Over a threeyear period from 2019 to 2022, a total of 96 specimens representing 15 species from four families were collected using standard entomological methods, including net sweeping and baiting across eight different locales. These families include Nymphalidae (3 species), Lycaenidae (4 species), Pieridae (6 species), and Papilionoidea (2 species). Notably, four species - Papilio demodocus, Belenois aurota, Colotis phisadia, and Colotis amata - are reported from Qatar for the first time, expanding the known butterfly fauna of the country. By amalgamating these novel findings with previously published results, this study presents an updated checklist of Qatar’s butterfly fauna, totalling 24 species spanning five families. This inventory sheds light on the diversity and distribution of butterflies in Qatar, emphasizing their resilience to extreme environmental conditions and underscoring the importance of conservation efforts amidst ongoing urbanization and agricultural expansion.

Phytosanitary state of open-field vegetable crop agroecosystems of the genus Solanum, Raphanus, Brassica in the central part of the Right-Bank Forest-Steppe of Ukraine

Goal. Phytosanitary assessment and determination of pest and disease prevalence and development in open-field vegetable crop plantings of the genus Solanum (tomato, pepper, eggplant), genus Raphanus (radish, oilseed radish, common radish, black radish, daikon), and genus Brassica (cabbage, cauliflower, broccoli) in the Central Part of the Right-Bank Forest-Steppe Region of Ukraine.&#x0D; Methods. The research was conducted using established entomological, phytopathological, and horticultural methods. From 2008 to 2022, the population, distribution, and development of pests and phytopathogens were assessed in households and small farms within the Cherkasy region of Ukraine.&#x0D; Results. Among the dominant pest species in the agroecosystems of vegetable crops from the genus Solanum, insects accounted for 80% (8 species), followed by mites at 10% (1 species), and slugs at 10% (1 species). In agroecosystems of Brassicaceae crops, insects caused significant damage, constituting 94% (16 species) of the total pest structure, along with naked snails (1 species). The area of infestation in tomato, sweet pepper, and eggplant crops by phytophagous insects ranged from 13% to 98%, with plant damage ranging from 14% to 65%. Common spider mites accounted for 26%. Exceedances of the Economic Pest Threshold (EPT) were observed, such as Leptinotarsa decemlineata Say at 2.6 times, Gryllotalpa gryllotalpa L., Trialeurodes vaporariorum Wstw., Agrotis segetum Denis &amp; Schiff., and Lacanobia oleracea at 2.2 times, Agriotes spp. larvae at 1.4 times, and Tetranychus urticae Koch. at 1.9 times the threshold levels. The highest area of infestation in Brassicaceae crops was found for Eurydema ventralis (46.7%), Pieris brassicae (42.4%), Agrotis segetum Denis &amp; Schiff. (41.6%), Phyllotreta crusiferae and Phyllotreta undulate (37.6—37.8%), and Brevicoryne brassicae (26.6%). Exceedances of the EPT were recorded at 1.1—2.5 times for these pests. In the structure of the phytopathogenic complex of vegetable crop agroecosystems, fungi were dominant, accounting for an average of 49—58%. Viral pathogens had the smallest share, approximately 6—8%, causing mosaic diseases. Bacterial and oomycete agents occupied an intermediate position in the pathogenic complex structure. Nevertheless, their negative impact on plant health could be significant.&#x0D; Conclusions. In the Cherkasy region, which encompasses the central part of the Right-Bank Forest-Steppe of Ukraine, approximately 75% of surveyed vegetable crop plantings, including tomatoes, peppers, eggplants, radishes, oilseed radishes, common radishes, black radishes, daikons, cabbages, cauliflowers, and broccolis, exhibit high pest infestations exceeding the EPT thresholds, indicating an ecologically hazardous phytosanitary state. In these agroecosystems, insects dominated, representing 80% and 94% of the pest structure in Solanum and Brassicaceae crops, respectively. Additionally, significant damage was attributed to naked snails (Kailie gliemeži) and, in the case of Solanum crops, mites (Tetranychus urticae Koch.). EPT exceedances ranged from 1.1 to 2.5 times, depending on the pest species. Fungi, causing mycoses, dominated the phytopathogenic complex structure, ranging from 14% to 40% (max 88—90%). The prevalence of diseases varied from 21% to 38%, with disease development rates ranging from 21% to 33%. Among the identified phytopathogens, five species are among the world’s most dangerous: Pseudomonas syringae and Xanthomonas campestris (bacteria), Botrytis cinerea and Fusarium oxysporum (fungi), and Phytophthora infestans (oomycete), which require strict control of their populations.

On the identification of the omnivorous humpback fly &lt;i&gt;Megaselia scalaris (Loew)&lt;/i&gt;

Ant-decapitating flies (lat. Megaselia scalaris (Loew)) is a quarantine object in the Russian Federation and is included in the Single list of quarantine objects of the Eurasian Economic Union, approved by the of the Council of the Eurasian Economic Commission dated 30.11.2016 No. 158, because it is a vector of dangerous diseases that cause great economic damage. The range covers large areas of North America, Africa, southern Europe, Australia, Southeast Asia, as well as the European part of Russia, namely the Southern Federal District and the Republic of Crimea. The list of regulated products subject to research to identify M. scalaris (Loew) is very extensive, and accurate identification to species of the received samples by entomological method by morphological features of copulatory organs is very labor-intensive and not accurate. Therefore, methods of molecular diagnostics were developed for identification, namely, comparison of the studied nucleotide sequence with the reference one using NCBI database and determination of genetic distances. The studies were carried out on the basis of the Department of Molecular Research of the North-West Testing Laboratory of FGBU «ARRIAH». Six samples of the genus Megaselia were used in this work. At the first stage, DNA was isolated using DNA-Extran 2 kits. Then genetic markers were obtained by classical PCR with subsequent detection by electrophoresis using the T100 Thermal Cycler, sequenced on a 3500 Genetic Analyzer. Identification was performed by comparing the obtained sequence in BioEdit program using NCBI database and by calculating genetic distances using two-parameter Kimura model and Tajima-Nei model. The study revealed advantages and disadvantages of the presented methods of molecular identification.

Design and Evaluation of a Sticky Attractant Trap for Intra-Domiciliary Surveillance of Aedes aegypti Populations in Mexico.

Surveillance consists of systematic data collection, analysis, and interpretation and is essential for planning and implementing control activities. The lack of success in the control and surveillance of the Ae. aegypti mosquito elsewhere demands the development of new accessible and effective strategies. This work aimed to develop and evaluate an adhesive lure trap for household indoor surveillance of Ae. aegypti. Based on a bibliographic review, four compounds that have significant attraction percentages for Ae. aegypti were considered. Our more effective blend was determined through preliminary bioassays using the high-throughput screening system (HITSS) and 90 × 90 cm mosquito cages. We designed a low-cost, pyramid-shaped, sticky cardboard trap to incorporate the selected blend. Semi-field 2 × 2 m cages and field tests were utilized to evaluate its effectiveness through mosquito capture percentages. In laboratory tests, blend number 2 presented an attraction percentage of 47.5 ± 4.8%; meanwhile, in semi-field cages, a 4-inch, 110 v powered fan was used to disperse the attractants, and then a similar capture percentage of 43.2 ± 4.0% was recorded. Results were recorded during the field evaluation of the at-house indoor environment and were compared with those recorded with the golden-standard BG-Sentinel trap, i.e., our prototype trapped an average of 6.0 ± 1.5 mosquitoes versus 10.0 ± 2.6. In most Latin American countries, there is a lack of formal and accessible strategies for monitoring adult populations of Ae. Aegypti; therefore, we must develop tools that reinforce entomological surveillance methods.

Field validation of post-mortem interval estimation based on insect development. Part 1: Accuracy gains from the laboratory rearing of insect evidence

There is a consensus that forensic methods must be valid. The high quality of the method may be fully demonstrated only through validation. Unfortunately, there are very few experimental or casework validations of entomological methods for estimating post-mortem interval (PMI). Here, we present the first part of the results from the field validation of minimum PMI (PMImin) estimates based on insect development. From eight pig carcasses (24–46.4 kg) exposed in a forest habitat of Western Poland, one every one or two months, we collected insect evidence with standard entomological techniques. Using weather station temperatures and the thermal summation method, PMImin was estimated based on insect life stages that were reared under controlled laboratory conditions. Through rearing an insect until the next developmental landmark (or until eclosion), its age at the time of collection (and eventually PMImin) may be estimated by subtracting thermal accumulation in the laboratory from the corresponding thermal constant. We hypothesized that rearing insect evidence significantly improves the accuracy of PMImin compared to the estimation based on non-reared insect evidence. The results clearly supported this hypothesis. However, the accuracy significantly increased only in the case of these insects that developed normally during rearing. When their development in the laboratory was prolonged, PMImin was significantly less accurate. For the normal development sample the accuracy improved in all species and life stages. The largest accuracy gains from rearing were recorded for Calliphora vomitoria (Diptera: Calliphoridae) and Stearibia nigriceps (Diptera: Piophilidae). Moreover, when puparia or third instar larvae were reared, gains were larger than in the case of earlier life stages. In conclusion, this study demonstrated that the method of rearing insect evidence to improve the accuracy of PMImin is valid. However, it needs to be used with caution, since substantial part of the evidence may die or slow down their development in the laboratory, which violates assumptions of the method.

Implication of agricultural practices in the micro-geographic heterogeneity of malaria transmission in Bouna, Côte d’Ivoire

BackgroundWetlands and irrigated agricultural crops create potential breeding sites for Anopheles mosquitoes, leading to a heterogeneity in malaria transmission. In agricultural areas, heterogeneity of malaria transmission is often associated with the presence of hotspots consisting of localized clusters of higher transmission intensity. This study aims to identify micro-geographic hotspots of malaria transmission in an agricultural setting using a multidisciplinary approach.MethodsTwo cross-sectional surveys were conducted at the end of the dry season and at the peak of the rainy season in rural and urban sites in Bouna, northeastern Côte d'Ivoire. A total of 296 individuals from 148 farming households were randomly selected and sociological, geographical, entomological, and clinical data as well as blood samples were collected during each visit. Parasitological data and Anopheles exposure (measured using entomological and immunological methods) were compared with demographic, agricultural, and geographic data to identify drivers of malaria transmission. Heat maps combining these data were used to identify households with ongoing malaria transmission throughout the year.ResultsIn rural areas, Plasmodium prevalence was consistent between the dry and the rainy seasons, with roughly half of the population infected. In urban areas, malaria transmission indicators were lower, with a parasite prevalence of less than 20%, which remained comparable between the dry and the rainy season. The presence of irrigated crops and proximity to wetlands were associated with increased Anopheles exposure. By mapping Plasmodium infection and Anopheles exposure, two different types of hotspots of malaria transmission were identified: micro-geographical scale and local scale hotspots.ConclusionsThe presence of wetlands in urban areas and irrigated agriculture in rural areas resulted in heterogeneity in malaria transmission on a micro-geographical scale. These specific households present particular risk of malaria transmission and could fuel malaria transmission in surrounding households. The identification of micro-geographical areas using heat maps combining several epidemiological parameters can help to identify hotspots of malaria transmission. The implementation of malaria control measures, such as seasonal chemoprophylaxis or vector control, in these areas could help to reduce the incidence of malaria and facilitate its elimination.

The Water‐Exclusion Trap (WET): A 3D printable window trap collector that prevents DNA degradation

Abstract Sampling is one of the most important and fundamental processes in field biology. Among many sampling methods in entomology, window traps have long been considered one of the most effective and standardized methods. Unfortunately, the DNA quality of samples collected by window traps is typically low due to incursion by water, mainly via rainfall. Water cannot be avoided in long‐term surveys, even with a roof, due to wind and condensation. As we move into the genomics era, this shortfall in DNA quality has become problematic for molecular biomonitoring, and solutions are urgently needed. In this study, we developed the Water‐Exclusion Trap (WET), a 3D printable long‐term storable bottom collector for window or pitfall traps. The collection medium in WETs does not dilute because water is excluded and evaporation is limited, maximizing DNA preservation. Our field tests show that the WET outperforms conventional traps in preserving DNA quality. Insect samples preserved in WETs in the field had significantly higher PCR success rates and DNA Integrity Numbers compared to conventional traps. The WET addresses a critical challenge in the use of window traps, DNA degradation caused by water intrusion, and represents a significant advancement in fieldwork for genomic research in entomology. We provide the 3D printing template of the WET, enabling immediate, free use for all. Furthermore, it can be easily customized for different environments and purposes, and can be adapted to a variety of other traps, including pitfall traps, funnel traps and more.