{"title":"Toxicity of spray adjuvants and tank mix combinations used in almond orchards to adult honey bees (Apis mellifera).","authors":"Brandon Shannon, Emily Walker, Reed M Johnson","doi":"10.1093/jee/toad161","DOIUrl":"10.1093/jee/toad161","url":null,"abstract":"<p><p>Commercial beekeepers transporting honey bees across the United States to provide almond pollination services have reported honey bee deaths, possibly due to pesticide applications made during crop bloom. Pesticides are often applied as \"tank mixes\", or mixtures of fungicides and insecticides combined into a single application. Spray adjuvants are often added to tank mixes to improve the application characteristics of a pesticide and include spreaders, stickers, or surfactants. The goal of this research was to determine toxicity of adjuvants to adult worker honey bees, both when applied alone and in adjuvant-pesticide tank mixtures. Field-relevant combinations of formulated products were applied to 3-day-old adult worker honey bees using a Potter Spray Tower, and mortality was assessed 48 h following exposure. Adjuvants tested included Activator-90, Attach, Choice Weather Master, Cohere, Dyne-Amic, Induce, Kinetic, LI 700, Liberate, Nu-Film P, PHT Latron B-1956, and Surf-90; fungicides tested include Luna Sensation (Fluopyram and Trifloxystrobin), Pristine (Pyraclostrobin and Boscalid), Tilt (Propiconazole), and Vangard (Cyprodinil), and insecticides tested include Altacor (Chlorantraniliprole), Intrepid 2F (Methoxyfenozide), and a positive control Mustang Maxx (Zeta-cypermethrin). Results demonstrated that exposure to some adjuvants causes acute honey bee mortality at near-field application rates, both when applied alone and in combination with pesticides. Some adjuvant-pesticide combinations demonstrated increased toxicity compared with the adjuvant alone, while others demonstrated decreased toxicity. A better understanding of adjuvant and adjuvant-pesticide tank mixture toxicity to honey bees will play a key role in informing \"Best Management Practices\" for pesticide applicators using spray adjuvants during bloom when honey bee exposure is likely.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1467-1480"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10564267/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10137258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sebastián Yánez-Segovia, Claudio C Ramírez, Richard L Lindroth, Eduardo Fuentes-Contreras
{"title":"Resistance against Leucoptera sinuella (Lepidoptera: Lyonetiidae) among hybrid clones of Populus spp. in central Chile.","authors":"Sebastián Yánez-Segovia, Claudio C Ramírez, Richard L Lindroth, Eduardo Fuentes-Contreras","doi":"10.1093/jee/toad129","DOIUrl":"10.1093/jee/toad129","url":null,"abstract":"<p><p>Leucoptera sinuella (Reutti) (Lepidoptera: Lyonetiidae) is a leaf miner specialist on Salicaceae recently introduced to Chile and Argentina, where it is causing economic damage to poplar plantations. We report a field survey in a poplar nursery naturally infested showing that regardless of the poplar hybrid taxon, high variability in resistance was observed among clones within families for oviposition and leaf-mining damage. A group of susceptible and resistant hybrid poplar clones was then selected for a laboratory evaluation of oviposition (antixenosis) and leaf-mining damage (antibiosis) on potted, rooted shoot cuttings. The concentration of condensed tannins (CTs) and salicinoid phenolic glucosides (SPGs) of the leaves of the selected clones from the laboratory study was also measured. Total oviposited eggs were positively correlated with leaf area, with the lowest oviposition on TMxT 11372 clone. The lowest percentage of mined leaf area was obtained for clones TMxT 11372, TMxT 11463, and TDxD 17574, but surprisingly no correlation between the percentage of mined leaf area and concentration of CTs and SPGs was found. Resistant poplar hybrids of our study could be suitable for breeding programs aimed for L. sinuella integrated pest management.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1662-1670"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9773057","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tim B Bryant, Jeremy K Greene, Francis P F Reay-Jones
{"title":"Within-field spatial patterns of Helicoverpa zea (Lepidoptera: Noctuidae) and spatial associations with stink bugs and their injury in field corn.","authors":"Tim B Bryant, Jeremy K Greene, Francis P F Reay-Jones","doi":"10.1093/jee/toad164","DOIUrl":"10.1093/jee/toad164","url":null,"abstract":"<p><p>The corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is a cosmopolitan pest in the field crop landscape in the southeastern United States. Field corn (Zea mays L.) is the most important midseason host for H. zea where intensive selection pressure occurs for resistance to insecticidal toxins from Bacillus thuringiensis (Bt). Because spatial patterns of H. zea in field corn have not been extensively studied, field corn was sampled for H. zea larvae and injury in 2021 and 2022. Patterns of spatial aggregation were identified in a number of fields in both larval populations and injury. Aggregation of H. zea larvae was less common at R5 than at R2. Associations between the spatial patterns of H. zea and the variability in crop phenology were identified in some fields, with positive associations between plant height and H. zea larvae, indicating that ovipositing H. zea moths avoid areas with reduced plant height and delayed reproductive maturity. Additionally, negative spatial associations between stink bug ear injury and H. zea larvae and their injury were found in a small number of cases, indicating some spatial interactions between the two pest complexes and their injury. Results from these studies provide valuable insight into the spatial patterns of H. zea in field corn. An understanding of the local dispersal and population dynamics of H. zea can be used to help further improve integrated pest management and insect resistance management programs for this major polyphagous pest.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1649-1661"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10395523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The effect of sunflower pollen age and origin on pathogen infection in the common eastern bumble bee (Apidae: Hymenoptera).","authors":"Alison E Fowler, Elisa Kola, Lynn S Adler","doi":"10.1093/jee/toad154","DOIUrl":"10.1093/jee/toad154","url":null,"abstract":"<p><p>Bumble bees are globally important pollinators, contributing hundreds of millions of dollars annually in crop pollination services. Several species are in decline, making it paramount to understand how pathogens and nutrition shape bee health. Previous work has shown that consuming sunflower pollen (Helianthus annuus) dramatically reduces infection by the trypanosomatid gut pathogen, Crithidia bombi, in the common eastern bumble bee (Bombus impatiens). Sunflower pollen may therefore be useful as a dietary supplement for reducing this pathogen in managed bumble bee colonies. Here, we assessed the efficacy of freezer-stored sunflower pollen that was collected in different years and locations for reducing pathogen infection. We tested sunflower pollen that was 1, 3, 4, or 5 yr old and from sunflowers grown in the United States or China against a control of 1-yr-old buckwheat pollen from China, since buckwheat pollen results in high infection. We hypothesized that older pollen would have weaker medicinal effects due to degradation of pollen quality. We found that all sunflower pollen treatments significantly decreased Crithidia infection compared to controls. These results suggest that sunflower pollen can be freezer-stored for up to 5 yr and sourced from a wide range of geographic areas and still maintain its medicinal effects against Crithidia in the common eastern bumble bee. This is helpful information for stakeholders who might administer sunflower pollen as a dietary supplement to manage Crithidia in commercial bumble bee colonies.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1939-1942"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9922938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lang Liu, Suji Wang, Junfeng Zuo, Xiaohe Zhang, Xiong Peng, Kang Wang, Maohua Chen
{"title":"Characterization and fitness cost of bifenthrin resistance in Rhopalosiphum padi (Hemiptera: Aphididae).","authors":"Lang Liu, Suji Wang, Junfeng Zuo, Xiaohe Zhang, Xiong Peng, Kang Wang, Maohua Chen","doi":"10.1093/jee/toad143","DOIUrl":"10.1093/jee/toad143","url":null,"abstract":"<p><p>Rhopalosiphum padi is an important global wheat pest. The pyrethroid insecticide bifenthrin is widely used in the control R. padi. We explored the resistance potential, cross-resistance, adaptive costs, and resistance mechanism of R. padi to bifenthrin using a bifenthrin-resistant strain (Rp-BIF) established in laboratory. The Rp-BIF strain developed extremely high resistance against bifenthrin (1033.036-fold). Cross-resistance analyses showed that the Rp-BIF strain had an extremely high level of cross-resistance to deltamethrin (974.483-fold), moderate levels of cross-resistance to chlorfenapyr (34.051-fold), isoprocarb (27.415-fold), imidacloprid (14.819-fold), and thiamethoxam (11.228-fold), whereas negative cross-resistance was observed to chlorpyrifos (0.379-fold). The enzymatic activity results suggested that P450 played an important role in bifenthrin resistance. A super-kdr mutation (M918L) of voltage-gated sodium channel (VGSC) was found in the bifenthrin-resistant individuals. When compared with the susceptible strain (Rp-SS), the Rp-BIF strain was significantly inferior in multiple life table parameters, exhibiting a relative fitness of 0.69. Our toxicological and biochemical studies indicated that multiple mechanisms of resistance might be involved in the resistance trait. Our results provide insight into the bifenthrin resistance of R. padi and can contribute to improve management of bifenthrin-resistant R. padi in the field.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1795-1803"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9851684","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Differential development and survival of Blattella asahinai and Blattella germanica (Blattodea: Ectobiidae) at six constant temperatures.","authors":"Madison K Peterson, Xing Ping Hu, Arthur G Appel","doi":"10.1093/jee/toad141","DOIUrl":"10.1093/jee/toad141","url":null,"abstract":"<p><p>The Asian cockroach, Blattella asahinai Mizukubo, is a peridomestic nuisance pest in the southeastern United States. Blattella asahinai is the closest relative to Blattella germanica (L.), the German cockroach, one of the most prolific and widespread domestic pests. Because these two species live in different habitats, they are expected to have differential development patterns reflecting environmental adaptations. Development of B. asahinai and B. germanica cockroach nymphs were observed at six constant temperatures ranging from 10 to 35 °C. At 10 °C and 15 °C, all nymphs died in the first instar, but B. germanica nymphs survived longer (10 °C: 12.9 d; 15 °C: 42.9 d) than B. asahinai nymphs (10 °C 8.2 d; 15 °C 18.4 d) at both temperatures. At 20 °C, 25 °C, and 30 °C, B. asahinai consistently had more instars and longer stadia than B. germanica. At 35 °C, only B. germanica was able to complete nymphal development; cannibalism among B. asahinai nymphs during molting was often observed at this temperature. The results for B. asahinai corroborated previously estimated growth patterns. The lower nymphal development threshold was 14.1 °C for B. germanica and 13.7 °C for B. asahinai. Comparing the development of B. germanica directly with its closest relative reveals specific physiological adaptations that B. germanica has developed for the indoor biome.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1776-1786"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9851689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yves Carrière, Ben Degain, Gopalan C Unnithan, Bruce E Tabashnik
{"title":"Inheritance and fitness cost of laboratory-selected resistance to Vip3Aa in Helicoverpa zea (Lepidoptera: Noctuidae).","authors":"Yves Carrière, Ben Degain, Gopalan C Unnithan, Bruce E Tabashnik","doi":"10.1093/jee/toad145","DOIUrl":"10.1093/jee/toad145","url":null,"abstract":"<p><p>The polyphagous pest Helicoverpa zea (Lepidoptera: Noctuidae) has evolved practical resistance to transgenic corn and cotton producing Cry1 and Cry2 crystal proteins from Bacillus thuringiensis (Bt) in several regions of the United States. However, the Bt vegetative insecticidal protein Vip3Aa produced by Bt corn and cotton remains effective against this pest. To advance knowledge of resistance to Vip3Aa, we selected a strain of H. zea for resistance to Vip3Aa in the laboratory. After 28 generations of continuous selection, the resistance ratio was 267 for the selected strain (GA-R3) relative to a strain not selected with Vip3Aa (GA). Resistance was autosomal and almost completely recessive at a concentration killing all individuals from GA. Declines in resistance in heterogeneous strains containing a mixture of susceptible and resistant individuals reared in the absence of Vip3Aa indicate a fitness cost was associated with resistance. Previously reported cases of laboratory-selected resistance to Vip3Aa in lepidopteran pests often show partially or completely recessive resistance at high concentrations and fitness costs. Abundant refuges of non-Bt host plants can maximize the benefits of such costs for sustaining the efficacy of Vip3Aa against target pests.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1804-1811"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10014108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yang Shi, Li He, Wenbing Ding, Hong Huang, Hualiang He, Jin Xue, Qiao Gao, Zhixiang Zhang, Youzhi Li, Lin Qiu
{"title":"Function analysis of CYP321A9 from Spodoptera frugiperda (Lepidoptera: Noctuidae) associated with emamectin benzoate, and a novel insecticide, cyproflanilide detoxification.","authors":"Yang Shi, Li He, Wenbing Ding, Hong Huang, Hualiang He, Jin Xue, Qiao Gao, Zhixiang Zhang, Youzhi Li, Lin Qiu","doi":"10.1093/jee/toad168","DOIUrl":"10.1093/jee/toad168","url":null,"abstract":"<p><p>The fall armyworm, Spodoptera frugiperda, is an invasive agricultural pest that is a serious threat to agricultural production and global food security. Chemical control is the most effective method for preventing outbreaks of S. frugiperda. However, insecticide resistance often develops as a result of prolonged pesticide use, and the molecular mechanisms involved in insecticide resistance remain unclear. Insect cytochrome P450 monooxygenases play an important role in the detoxification of insecticides and insecticide resistance in Lepidoptera. In our study, the LC50 of a novel insecticide (cyproflanilide) and a conventional insecticide (emamectin benzoate) for S. frugiperda second-instar larvae were 7.04 and 1.61 mg/L, respectively. Furthermore, CYP321A9 expression was upregulated in larvae exposed to these insecticides. Additionally, knockdown of CYP321A9 by feeding larvae with dsRNA for 72 h significantly increased the mortality of S. frugiperda exposed to emamectin benzoate and cyproflanilide by 23.33% and 7.78%, respectively. Our results indicate that CYP321A9 may play an important role in the detoxification of emamectin benzoate and cyproflanilide in S. frugiperda. Our findings provide a basis to better understand the mechanisms of insecticide resistance and contribute to the control of S. frugiperda.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1812-1819"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10119256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhengyan Wang, Haisheng Hu, Zhenzhen Chang, Shan Zhang, Yujie Lu
{"title":"Molecular mechanism of Enterococcus faecalis-induced phosphine sensitivity in Tribolium castaneum (Coleoptera: Tenebrionidae).","authors":"Zhengyan Wang, Haisheng Hu, Zhenzhen Chang, Shan Zhang, Yujie Lu","doi":"10.1093/jee/toad171","DOIUrl":"10.1093/jee/toad171","url":null,"abstract":"<p><p>Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) has developed extensive resistance to the fumigant phosphine. Knowledge of the resistance mechanisms offers insight into resistance management. Although several studies have highlighted the positive or negative impacts of symbiotic microbiota on host pesticide resistance, little is known about the association between gut symbionts and host phosphine resistance. To reveal the effect of the gut bacterium, Enterococcus faecalis (Andrewes & Horder) (Lactobacillales: Enterococcaceae), on host phosphine resistance and its underlying mechanism, we investigated mortality, fitness, redox responses, and immune responses of adult T. castaneum when challenged with E. faecalis inoculation and/or phosphine exposure. When T. castaneum was exposed to phosphine, E. faecalis inoculation decreased its survival and female fecundity and aggravated its oxidative stress. Furthermore, E. faecalis inoculation suppressed the expression and activity of superoxide dismutase, catalase, and peroxidase in phosphine-exposed T. castaneum. Enterococcus faecalis inoculation also triggered excessive host immune responses, including the immune deficiency signaling pathway and the dual oxidase-reactive oxygen species system. These findings suggest that E. faecalis likely modulates host phosphine resistance by interfering with the redox system. This provides information for examining the symbiotic function in the insect-microorganism relationship and new avenues for pesticide resistance management.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1820-1829"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10123456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Genetic differentiation and species diversification of the Adoxophyes orana complex (Lepidoptera: Tortricidae) in East Asia.","authors":"Cheng-Lung Tsai, Li-Cheng Shih, Wen-Bin Yeh, Bong-Kyu Byun, Utsugi Jinbo, Fang-Yu Ning, I-Hsin Sung","doi":"10.1093/jee/toad155","DOIUrl":"10.1093/jee/toad155","url":null,"abstract":"<p><p>The Adoxophyes tea tortrix (Lepidoptera: Tortricidae) is a group of leaf rollers that cause enormous economic losses on tea and apple crops. In East Asia, taxonomic ambiguity of the Adoxophyes orana complex (AOC), which consists of A. orana, A. dubia, A. honmai, and A. paraorana, has persisted for decades because of vague diagnostic characters. In this study, differences in the AOC were examined to improve species identification, determine genetic variations, and develop control strategies. Analyses revealed that A. orana comprised 2 lineages, a widely distributed Palearctic lineage and an East Asian lineage that was nested with other Asian species. Genetic divergence of >3% is proposed to confirm the AOC species that would benefit subsequent taxonomic revision. The monophyletic Taiwanese A. sp. with 2.8-4% from other AOC species appeared to suggest it as an independent taxon, and low interspecific divergence between A. honmai and A. dubia of 0.3% indicated possibility of recent divergence or intraspecific variations. Our result further suggested that the Z9-14:Ac ratio of semiochemicals could be a reference for the reblending of pheromone attractants in Taiwanese tea plantations. Moreover, the AOC species appeared to have a tendency of specific geographic distributions, with A. dubia and A. honmai in Japan and China, A. paraorana in Korea, and A. sp. in Taiwan. Maintaining the unique genetic composition of Adoxophyes species in each geographic region and preventing the possible invasions into those AOC-free countries through the transportation of host plants are essential in managing the AOC in East Asia.</p>","PeriodicalId":15632,"journal":{"name":"Journal of Economic Entomology","volume":" ","pages":"1885-1893"},"PeriodicalIF":2.2,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10297738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}