Polyandry may mitigate the negative impact of reproductive interference among bumblebees in Japan.

In social hymenopterans, monandry of the queen is an ancestral trait, and polyandry is a derived trait. Polyandry of the queen is the norm in a limited number of lineages, such as honeybees, leaf-cutting ants, Pogonomyrmex ants, and Vespula wasps, which presumably provide fitness advantages for the whole colony. The queen of the introduced bumblebee, Bombus terrestris, is polyandrous in Japan, whereas it is monandrous in native regions. We hypothesize that polyandry can evolve in a process that avoids the negative impacts of reproductive interference caused by interspecific mating and conducted genetic studies of the invasive species B. terrestris and two native subspecies, Bombus hypocrita sapporoensis and Bombus hypocrita hypocrita, in Japan. Our results revealed that although the native queens of B. hypocrita hypocrita allopatric with B. terrestris were strictly monandrous, the native queens of B. hypocrita sapporoensis sympatric with B. terrestris were polyandrous. These results suggested that the queens of native B. hypocrita sapporoensis do not experience negative impacts on interspecific mating from the invasive B. terrestris. We discuss the possibility that reproductive interference is a driving force in selection for multiple mating through an arms race between sympatric species.

Novel approach for detecting classical swine fever virus from swabs of wild boar cut tails using nested real-time PCR.

We devised a method to detect the classical swine fever virus (CSFV) in tail-wiped swabs from wild boars. The CSFV gene in swabs was detected with high sensitivity using nested real-time polymerase chain reaction (PCR), which is a combination of reverse transcription-PCR (RT-PCR) and real-time PCR. We compared CSFV gene detection from boar tissue using the conventional and our tail-wiped swab method. The tail-wiped swab method showed sensitivity and specificity of 100% (26/26) and 98.8% (172/174), respectively compared to the conventional method. Thus, the swab-based CSFV detection method was considered to have detection sensitivity comparable to that of conventional methods. Additionally, we conducted surveillance for CSFV in wild boars on Awaji Island. CSFV was detected in 10.7% (45/420) of samples.

Sequencing methods for HA and NA genes of avian influenza viruses from wild bird feces using Oxford Nanopore sequencing.

We developed a method to determine the sequences of hemagglutinin (HA) and neuraminidase (NA) from RNA extracted directly from wild bird fecal samples, using Nanopore Flongle. We determined the nucleotide sequences and subtypes of HA and NA in 16 and 15 samples respectively, using Flongle. The results of HA and NA subtyping determined by the conventional method were consistent with their subtypes determined by our method, thereby the applicability of this method in the identification of HA and NA subtypes. In addition, the homology between the HA fragments in this and the Sanger methods ranged from 98.5 % to 100 %. Compared with conventional PCR with the Sanger method, this method can easily determine HA and NA subtypes and sequences directly from the fecal samples. It is easier to implement and has lower running costs (USD100$) than other NGS-based methods, making it a useful tool for avian influenza surveillance in wild birds.

Insecticide Residues Associated with Apple Orchard Treatments in the Mason Bee, Osmia cornifrons, and their Nests.

Declines in bee populations, in part due to pesticides, especially insecticides, are of global concern. Although most studies have investigated insecticide residues in honeybees and bumblebees, few have focused on non-Apis solitary bees, which are considered essential pollinators in the field. Most non-Apis bees are solitary and build their nests in the ground or in tree holes, therefore insecticide exposure pathways would differ from those of honeybees and bumblebees. We analyzed the residues of 20 insecticides in Osmia cornifrons bees and their pollen-provisions and nesting materials, along with adult honeybees, soil, and wildflowers collected in four apple orchards in two regions in Japan. Few insecticides were common among adult bees, pollen-provisions, and wildflowers. Insecticides applied in orchards were detected at high frequency: chlorantraniliprole, flubendiamide, and diazinon were detected in almost all samples. Insecticides without a known history of application were detected from various samples at frequencies ranging from 0% to 100%. Even in orchards without a known history of insecticide application, clothianidin was detected in many sample types and at high concentration. A purple deadnettle sample had the highest concentration at 17.5 mg/kg. These results highlight the complexity of pathways of insecticide exposure to O. cornifrons in the environment because insecticides may remain in the environment for long periods and wild bees may forage farther than previously known distances. Furthermore, each sample type contained different insecticides, so wild bees may have been exposed to different insecticides at different life stages. Future research should prioritize wide-scale measurements of insecticide residues in field conditions and toxicity testing with multiple exposures at different life stages of target species. Environ Toxicol Chem 2023;42:1564-1574. © 2023 SETAC.

Acute toxicity data of common agricultural insecticides to Japanese wild bees.

Although numerous ecotoxicological assessments of European honeybee (Apis mellifera L.) have been performed, Japanese wild bees are less well studied in this regard. To address this knowledge gap, we assessed the mortality and acute toxicity (LD50) of 3 common agricultural insecticides (clothianidin, fipronil, and diazinon) on as many as 6 species of Japanese wild bees (Andrena prostomias Perez, Apis cerana japonica Radoszkowski, Bombus deuteronymus Perez, Bombus honshuensis Tkalcu, Bombus hypocrita Perez, and Eucera spp.; all or any of them). The datasets were obtained via standard acute toxicity testing, with assessment of mortality at 24 and 48 h after exposure to the insecticides. These data provide important information regarding the effects of insecticides on Japanese wild bees and their conservation.

Efficiency of ant-control agents in colony-level oral toxicity tests using Tetramorium tsushimae (Hymenoptera: Formicidae) for post-establishment control of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae).

The red imported fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) causes serious damage worldwide as an invasive alien species. The species has expanded its range to the Pacific Rim since 2000s and Japan has faced its multiple introductions since 2017. While colony-level control methods are urgently needed, testing living colonies of the unestablished species is challenging especially due to various restrictions under the COVID-19 pandemic. Here, we proposed alternative long-term toxicity assays using artificial colonies of Tetramorium tsushimae Emery (Hymenoptera: Formicidae), a Japanese native species belonging to the same subfamily (Myrmicinae) as S. invicta. We conducted an acute toxicity test to determine if T. tsushimae is a suitable substitute for S. invicta using fipronil and found the LD50 value in T. tsushimae was close to that in S. invicta. Then, we conducted the long-term toxicity test with fipronil and two insect growth regulators (pyriproxyfen and etoxazole) using artificial colonies of T. tsushimae. All workers and larvae in the fipronil-treated colonies died within 3 days of treatment initiation. Emergence of new workers was observed after 18 days in the etoxazole-treated and control colonies, but not in the pyriproxyfen-treated colonies. We concluded that fipronil was the most promising insecticide for post-establishment control, and pyriproxyfen was effective as a toxic-bait agent for colony-level control. Supplementary Information: The online version contains supplementary material available at 10.1007/s13355-022-00800-x.

Effects of neonicotinoids on honey bee autogrooming behavior against the tracheal mite Acarapis woodi.

The European honey bee, Apis mellifera, is the most common and important pollinator of crops worldwide. Honey bees are damaged by destructive parasitic mites, but they also have evolved a behavioral immune system to remove them. Exposures to neonicotinoids, however, can cause significant behavioral effects because these compounds alter the central role of nicotinic acetylcholine receptor in insect brains. In this study, we assessed the effects of three neonicotinoids that have a high toxicity to bees-imidacloprid, thiamethoxam, and clothianidin-on the behavioral immune system of honey bees. We used A. mellifera and the endoparasitic mite Acarapis woodi as a behavioral immune system model because A. mellifera can effectively remove the mite by autogrooming. Our results did not demonstrate an effect of neonicotinoid application on whether bees show autogrooming or on mite removal, but the time to initial autogrooming became shorter and the number of autogrooming attempts increased. As opposed to previous studies, our findings indicate that the honey bee response to parasitic mites becomes more sensitive after exposure to neonicotinoids.Clinical Trials Registration: Not applicable.

Distribution and Genetic Diversity of the Amphibian Chytrid in Japan.

While research on frog chytrid fungus Batrachochytrium dendrobatidis (Bd), an infectious disease that threatens amphibian diversity, continues to advance worldwide, little progress has been made in Japan since around 2010. The reason for this is, which we pointed out in 2009, that the origin of frog chytrid fungus may be in the East Asian region, including Japan based on the Bd ITS-DNA variation, and as few cases of mass mortality caused by this fungus have been observed in wild amphibian populations in Japan, the interest of the Japanese government and the general public in Bd has waned. However, we believe that organizing the data obtained so far in Japan and distributing the status of frog chytrid fungus in Japan to the world will provide useful insight for future risk management of this pathogen. We collected more than 5500 swab samples from wild amphibians throughout Japan from 2009 to 2010. Then, we investigated the infection status using the Nested-PCR method. We sequenced the obtained DNA samples and constructed a maximum-parsimony (MP) tree to clarify the phylogenetic diversity of Bd. We detected Bd infection in 11 (nine native and two alien) amphibian species in Japan and obtained 44 haplotypes of Bd ITS-DNA. The MP tree showed a high diversity of Bd strains in Japan, suggesting that some strains belong to Bd-GPL and Bd-Brazil. Except for local populations of the Japanese giant salamanders Andrias japonicus in Honshu Island and the sword tail newts Cynops ensicauda in Okinawa Island, the Bd infection prevalence in native amphibian species was very low. The alien bullfrog Aquarana catesbeiana had high Bd infection rates in all areas where they were sampled. No Bd infection was detected in other native amphibians in the areas where giant salamanders, sword tail newts, and bullfrogs were collected, suggesting that many native amphibians are resistant to Bd infection. The sword tail newt of Okinawa Island had both the highest infectious incidence and greatest number of haplotypes. The giant salamanders also showed relatively high infection prevalence, but the infected strains were limited to those specific to this species. These two Caudata species are endemic to a limited area of Japan, and it was thought that they may have been refugia for Bd, which had been distributed in Japan Islands for a long time.

Epidemiologic Survey of Avian Influenza Virus Infection in Shorebirds Captured in Hokkaido, Japan.

There is limited information about virus epidemiology of shorebirds (family Charadriidae and Scolopacidae) in the East Asia-Australasia flyway. We investigated the prevalence of avian influenza viruses (AIVs) in shorebirds in Hokkaido, Japan, the stopover site of the flyway, to understand the ecology of AIV translocation in the flyway from 2006 to 2010. In total, 1,698 shorebirds belonging to 26 species were captured and released into two different sites using mist nets. Tracheal and cloacal swabs were collected from each bird using cotton swabs. The RNA of influenza A viruses was detected using reverse transcription loop-mediated isothermal amplification. One AIV-positive sample was obtained from a Lesser Sand Plover (Charadrius mongolus) captured in September 2010 at Lake Komuke. Full lengths of hemagglutinin (HA), neuraminidase (NA), polymerase acidic protein, nucleoprotein, matrix protein 1, and nuclear export protein genes were successfully amplified from the AIV-positive sample. All sequences showed the highest identity with sequences obtained from virus strains from Anseriformes species. Shorebirds migrated to Japan 1 mo earlier than did Anseriformes species. Therefore, the Lesser Sand Plover could have been infected by the virus from Anseriformes species on the breeding grounds. The HA sequence showed the highest identity with the H10 sequence whereas the NA sequence exhibited the highest identity with the N7 sequence. Phylogenic analysis showed that the detected subtype H10N7 belongs to the Eurasia lineage and the related strain might have widely spread in Asia in 2009.

Effects of a herbicide on paddy predatory insects depend on their microhabitat use and an insecticide application.

Indirect effects of agrochemicals on organisms via biotic interactions are less studied than direct chemical toxicity despite their potential relevance in agricultural landscapes. In particular, the role of species traits in characterizing indirect effects of pesticides has been largely overlooked. Moreover, it is still unclear whether such indirect effects on organisms are prevalent even when the organisms are exposed to direct toxicity. We conducted a mesocosm experiment to examine indirect effects of a herbicide (pentoxazone) on aquatic predatory insects of rice paddies. Because the herbicide selectively controls photosynthetic organisms, we assumed that the effects of the herbicide on predatory insects would be indirect. We hypothesized that phytophilous predators such as some Odonata larvae, which cling to aquatic macrophytes, would be more subject to negative indirect effects of the herbicide through a decrease in abundance of aquatic macrophytes than benthic, nektonic, and neustonic predators. Also, we crossed-applied an insecticide (fipronil) with herbicide application to examine whether the indirect effects of the herbicide on the assembling predators act additively with direct adverse effects of the insecticide. The herbicide application did not decrease the abundance of phytoplankton constitutively, and there were no clear negative impacts of the herbicide on zooplankton and prey insects (detritivores and herbivores). However, the abundance of aquatic macrophytes was significantly decreased by the herbicide application. Although indirect effects of the herbicide were not so strong on most predators, their magnitude and sign differed markedly among predator species. In particular, the abundance of phytophilous predators was more likely to decrease than that of benthic, nektonic, and neustonic predators when the herbicide was applied. However, these indirect effects of the herbicide could not be detected when the insecticide was also applied, seemingly due to fipronil's high lethal toxicity. Our study highlights the importance of species traits such as microhabitat use, which characterize biotic interactions, for predicting indirect effects of agrochemicals. Given that indirect effects of the chemicals vary in response to species traits and direct toxicity of other chemicals, efforts to explain this variation are needed to predict the realistic risks of indirect effects of agrochemicals in nature.

Ecological risks of insecticide contamination in water and sediment around off-farm irrigated rice paddy fields.

The ecological impacts of insecticides in aquatic areas around agricultural lands have long been ignored in the regulation scheme of pesticides in Japan. Upon the scheme, the predicted concentration of an insecticide in the main stream of a river is the only parameter considered, suggesting that the ecological impacts of insecticides on local biodiversity around agricultural fields are underestimated. To fill this knowledge gap, we measured insecticide concentrations in surface water and sediment in aquatic areas around paddy fields at 35 locations across Japan. Among the 18 insecticides considered, 15 were detected somewhere in Japan and their concentrations were generally higher in the southwestern region in Japan (e.g. Hiroshima, Saga, or Kagoshima prefectures). Most insecticides were accumulated at higher concentrations in sediment than in surface water, consistent with previous studies. We also detected insecticides applied to nursery boxes at high concentrations in surrounding aquatic areas, although such application is generally considered to have low environmental risks. In addition, derivatives of fipronil, which have similar toxicity as that of fipronil, were often detected in sediment at higher concentrations than fipronil itself. Concentrations of dinotefuran in water at two sampling points were higher than the 5% hazardous concentration (HC5), indicating a possibility of a risk of acute toxicity to aquatic organisms. Our findings indicate that ecological risk assessments of insecticides and their derivatives should be expanded to include concentrations in sediment and water around paddy fields as well.

Reproductive Interference in an Introduced Bumblebee: Polyandry may Mitigate Negative Reproductive Impact.

As a signature of reproductive interference (RI), we reviewed hybrid production in eusocial bumblebees in Japan, by comparing introduced Bombus terrestris with native B. ignitus in Honshu (main island of Japan) and with native B. hypocrita sapporoensis in Hokkaido (northern island of Japan). In this review, we present additional new data showing hybrid production between introduced B. terrestris and native B. ignitus in Honshu. Interspecific mating with introduced B. terrestris disrupts the reproduction of native B. h. sapporoensis and B. ignitus, which belong to the same subgenus of Bombus, through inviable egg production. This interference appears to facilitate species replacement on Hokkaido. Simultaneously, the mating frequencies for queens of B. terrestris have increased, suggesting that polyandry might evolve in response to the extent of RI between B. terrestris and B. h. sapporoensis. To suppress the population size of B. terrestris in Hokkaido, two methods have been proposed: the mass release of B. h. sapporoensis males to induce RI between the two species and the spraying of insecticides against foraging workers so that the workers will carry the insecticides back to their colonies, killing the immature bees within the colonies. A candidate insecticide type is insect growth regulator, which may disrupt larval development without any apparent effect on foraging workers.

Effects of short-term, sublethal fipronil and its metabolite on dragonfly feeding activity.

Dragonflies, Sympetrum spp., are indispensable to agriculture and are a central element of culture in Japan. However, S. frequens populations in rice paddy fields have declined in recent decades. Dragonfly larvae are predatory aquatic insects that feed on other organisms found in habitats with slow-moving or standing water. The increasing use of fipronil and neonicotinoid insecticides in agriculture is also increasing exposure to Sympetrum spp. in larval stages through paddy soil and water. The role of fipronil insecticides in the decline of dragonflies is of concern, and we here examine the sublethal effects of this insecticide on the feeding behaviors of two Sympetrum spp. Based on the quantity of prey items consumed and the time to capture prey items, feeding inhibition was determined to be a potential mechanism of the decline of Sympetrum spp. following 48-h exposure to fipronil and fipronil sulfone. Prey consumption by S. infuscatum was significantly reduced for fipronil sulfone at all concentrations (0.01-1000 µg/L). S. frequens exposed to 1, 10, 100 and 1000 µg/L fipronil sulfone had significantly longer prey capture times. Fipronil sulfone was 2.8, 9.7 and 10.5 times more toxic to S. infuscatum than fipronil in terms of acute toxicity, feeding inhibition and delayed toxicity, respectively. In addition, fipronil sulfone was 6.6, 2.9 and 9.1 times more toxic, respectively, to S. frequens than fipronil. Our findings suggest that sublethal effects on feeding inhibition lead to severe mortality at realistic paddy soil and water concentrations. Our results provide the first demonstration that short-term exposure to fipronil and fipronil sulfone can consequently cause significant harm to dragonfly larvae survival due to feeding inhibition. These findings have implications for current pesticide risk assessment and dragonfly protection.

Author Correction: Declaration of local chemical eradication of the Argentine ant: Bayesian estimation with a multinomial-mixture model.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Declaration of local chemical eradication of the Argentine ant: Bayesian estimation with a multinomial-mixture model.

Determining the success of eradication of an invasive species requires a way to decide when its risk of reoccurrence has become acceptably low. In Japan, the area populated by the Argentine ant, Linepithema humile (Mayr), is expanding, and eradication via chemical treatment is ongoing at various locations. One such program in Tokyo was apparently successful, because the ant population decreased to undetectable levels within a short time. However, construction of a population model for management purposes was difficult because the probability of detecting ants decreases rapidly as the population collapses. To predict the time when the ant was eradicated, we developed a multinomial-mixture model for chemical eradication based on monthly trapping data and the history of pesticide applications. We decided when to declare that eradication had been successful by considering both 'eradication' times, which we associated with eradication probabilities of 95% and 99%, and an optimal stopping time based on a 'minimum expected economic cost' that considered the possibility that surveys were stopped too soon. By applying these criteria, we retroactively declared that Argentine ants had been eradicated 38-42 months after the start of treatments (16-17 months after the last sighting).

Characterizing the temporal patterns of avian influenza virus introduction into Japan by migratory birds.

The objectives of the present study were to observe the temporal pattern of avian influenza virus (AIV) introduction into Japan and to determine which migratory birds play an important role in introducing AIV. In total, 19,407 fecal samples from migratory birds were collected at 52 sites between October 2008 and May 2015. Total nucleic acids extracted from the fecal samples were subjected to reverse transcription loop-mediated isothermal amplification to detect viral RNA. Species identification of host migratory birds was conducted by DNA barcoding for positive fecal samples. The total number of positive samples was 352 (prevalence, 1.8%). The highest prevalence was observed in autumn migration, and a decrease in prevalence was observed. During autumn migration, central to southern Japan showed a prevalence higher than the overall prevalence. Thus, the main AIV entry routes may involve crossing the Sea of Japan and entry through the Korean Peninsula. Species identification was successful in 221 of the 352 positive samples. Two major species sequences were identified: the Mallard/Eastern Spot-billed duck group (115 samples; 52.0%) and the Northern pintail (61 samples; 27.6%). To gain a better understanding of the ecology of AIV in Japan and the introduction pattern of highly pathogenic avian influenza viruses, information regarding AIV prevalence by species, the prevalence of hatch-year migratory birds, migration patterns and viral subtypes in fecal samples using egg inoculation and molecular-based methods in combination is required.

Insecticide Susceptibility in Asian Honey Bees (Apis cerana (Hymenoptera: Apidae)) and Implications for Wild Honey Bees in Asia.

To conserve local biodiversity and ensure the provision of pollination services, it is essential to understand the impact of pesticides on wild honey bees. Most studies that have investigated the effects of pesticides on honey bees have focused on the European honey bee (Apis mellifera (Hymenoptera: Apidae)), which is commonly domesticated worldwide. However, the Asian honey bee (Apis cerana) is widely distributed throughout Asia, and toxicity data are lacking for this species. This study aimed to fill this important knowledge gap. In this study, we determined the acute contact toxicity in A. cerana to various pesticides, including neonicotinoids, fipronil, organophosphorus, synthetic pyrethroids, carbamate, and anthranilic diamide. Based on the test duration of 48 h of contact LD50 tests, A. cerana was most sensitive to dinotefuran (0.0014 µg/bee), followed by thiamethoxam (0.0024 µg/bee) and fipronil (0.0025 µg/bee). Dinotefuran is used extensively in Asia, thereby potentially creating a substantial hazard. More generally, A. cerana was approximately one order of magnitude more sensitive than was A. mellifera to most of the pesticides evaluated. The results of our study suggest that neonicotinoid pesticides should not be considered as a single group that acts uniformly on all honey bees, and that more careful management strategies are required to conserve A. cerana populations than A. mellifera.

Genetic Structure and Potential Environmental Determinants of Local Genetic Diversity in Japanese Honeybees (Apis cerana japonica).

Declines in honeybee populations have been a recent concern. Although causes of the declines remain unclear, environmental factors may be responsible. We focused on the potential environmental determinants of local populations of wild honeybees, Apis cerana japonica, in Japan. This subspecies has little genetic variation in terms of its mitochondrial DNA sequences, and genetic variations at nuclear loci are as yet unknown. We estimated the genetic structure and environmental determinants of local genetic diversity in nuclear microsatellite genotypes of fathers and mothers, inferred from workers collected at 139 sites. The genotypes of fathers and mothers showed weak isolation by distance and negligible genetic structure. The local genetic diversity was high in central Japan, decreasing toward the peripheries, and depended on the climate and land use characteristics of the sites. The local genetic diversity decreased as the annual precipitation increased, and increased as the proportion of urban and paddy field areas increased. Positive effects of natural forest area, which have also been observed in terms of forager abundance in farms, were not detected with respect to the local genetic diversity. The findings suggest that A. cerana japonica forms a single population connected by gene flow in its main distributional range, and that climate and landscape properties potentially affect its local genetic diversity.

Spatial assessment of the potential risk of avian influenza A virus infection in three raptor species in Japan.

Avian influenza A, a highly pathogenic avian influenza, is a lethal infection in certain species of wild birds, including some endangered species. Raptors are susceptible to avian influenza, and spatial risk assessment of such species may be valuable for conservation planning. We used the maximum entropy approach to generate potential distribution models of three raptor species from presence-only data for the mountain hawk-eagle Nisaetus nipalensis, northern goshawk Accipiter gentilis and peregrine falcon Falco peregrinus, surveyed during the winter from 1996 to 2001. These potential distribution maps for raptors were superimposed on avian influenza A risk maps of Japan, created from data on incidence of the virus in wild birds throughout Japan from October 2010 to March 2011. The avian influenza A risk map for the mountain hawk-eagle showed that most regions of Japan had a low risk for avian influenza A. In contrast, the maps for the northern goshawk and peregrine falcon showed that their high-risk areas were distributed on the plains along the Sea of Japan and Pacific coast. We recommend enhanced surveillance for each raptor species in high-risk areas and immediate establishment of inspection systems. At the same time, ecological risk assessments that determine factors, such as the composition of prey species, and differential sensitivity of avian influenza A virus between bird species should provide multifaceted insights into the total risk assessment of endangered species.

Fipronil application on rice paddy fields reduces densities of common skimmer and scarlet skimmer.

Several reports suggested that rice seedling nursery-box application of some systemic insecticides (neonicotinoids and fipronil) is the cause of the decline in dragonfly species noted since the 1990s in Japan. We conducted paddy mesocosm experiments to investigate the effect of the systemic insecticides clothianidin, fipronil and chlorantraniliprole on rice paddy field biological communities. Concentrations of all insecticides in the paddy water were reduced to the limit of detection within 3 months after application. However, residuals of these insecticides in the paddy soil were detected throughout the experimental period. Plankton species were affected by clothianidin and chlorantraniliprole right after the applications, but they recovered after the concentrations decreased. On the other hand, the effects of fipronil treatment, especially on Odonata, were larger than those of any other treatment. The number of adult dragonflies completing eclosion was severely decreased in the fipronil treatment. These results suggest that the accumulation of these insecticides in paddy soil reduces biodiversity by eliminating dragonfly nymphs, which occupy a high trophic level in paddy fields.