Entomology Commons^™

Parameters For Successful Parental Rnai As An Insect Pest Management Tool In Western Corn Rootworm, Diabrotica Virgifera Virgifera, Ana Maria Vélez, Elane Fishilevich, Natalie Matz, Nicholas P. Storer, Kenneth E. Narva, Blair D. Siegfried

Department of Entomology: Faculty Publications

Parental RNAi (pRNAi) is an RNA interference response where the gene knockdown phenotype is observed in the progeny of the treated organism. pRNAi has been demonstrated in female western corn rootworms (WCR) via diet applications and has been described as a potential approach for rootworm pest management. However, it is not clear if plant-expressed pRNAi can provide effective control of next generation WCR larvae in the field. In this study, we evaluated parameters required to generate a successful pRNAi response in WCR for the genes brahma and hunchback. The parameters tested included a concentration response, duration of the dsRNA …

Insect And Plant-Derived Mirnas In Greenbug (Schizaphis Graminum) And Yellow Sugarcane Aphid (Sipha Flava) Revealed By Deep Sequencing, Haichuan Wang, Chi Zhang, Yongchao Dou, Bin Yu, Yunfeng Liu, Tiffany Heng-Moss, Guoqing Lu, Michael Wachholtz, Jeffery D. Bradshaw, Paul Twigg, Erin Scully, Nathan Palmer, Gautam Sarath

Department of Entomology: Faculty Publications

Schizaphis graminum (green bug; GB) and Sipha flava (yellowsugarcane aphid; YSA) are two cereal aphid species with broad host ranges capable of establishing on sorghum (Sorghum bicolor) and several switchgrass (Panicum virgatum) cultivars. Switchgrass and sorghum are staple renewable bioenergy crops that are vulnerable to damage by aphids, therefore, identifying novel targets to control aphids has the potential to drastically improve yields and reduce losses in these bioenergy crops. Despite thewealth of genomic and transcriptomic information available from a closely related model aphid species, the pea aphid (Acyrthosiphon pisum), similar genomic information, including the …

Fitness Costs Associated With Cry1f Resistance In The European Corn Borer, J. L. Petzold-Maxwell, Blair D. Siegfried, R. L. Hellmich, C. A. Abel, B. S. Coates, Terrence A. Spencer, R. J. Horikoshi, Aaron J. Gassmann

Department of Entomology: Faculty Publications

Crops producing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely planted to manage insect pests. Bt crops can provide an effective tool for pest management; however, the evolution of Bt resistance can diminish this benefit. The European corn borer, Ostrinia nubilalis H€ubner, is a significant pest of maize and is widely managed with Bt maize in the Midwest of the United States. When Bt crops are grown in conjunction with non-Bt refuges, fitness costs of Bt resistance can delay the evolution of resistance. Importantly, fitness costs often vary with ecological factors, including host-plant genotype and diapause. In …

Farmers’ Perspectives On Resistance In Western Corn Rootworm To Crw-Bt Corn In Midwest Usa, David A. Andow, Robert Wright, Erin W. Hodgson, Thomas E. Hunt, Ken Ostlie

Department of Entomology: Faculty Publications

Resistance in western corn rootworm to transgenic corn hybrids was first confirmed in 2011 in Midwestern USA, and threatens their continued use. Farmers are often the first line of resistance detection, so their understanding and attitudes toward this issue are critical for improving resistance management. We conducted telephone focus groups during 2013 with farmers who had experienced rootworm resistance. There were four stages in dealing with unexpected rootworm injury: Awareness of a problem, diagnosis, confirmation, and recommendations. Most farmers discovered the problem themselves, but this usually happened too late in the growing season to limit yield loss. Once aware of …

Obituary And Bibliography Of Norm Penny (1946-2016), An “Entomologist’S Entomologist”, Robert L. Zuparko, Brett C. Ratcliffe

Department of Entomology: Faculty Publications

The section “Remembrances of Norm Penny” was authored by BCR, and the remaining sections by RLZ. Fabio Penny and Rachel Diaz-Bastin made valuable additions to the text.

Norman Dale Penny, collection manager emeritus in the Department of Entomology at the California Academy of Sciences, San Francisco, California, and past president of the Pacifi c Coast Entomological Society, died of a stroke on July 10, 2016. That weekend he and his wife were near Quincy, in Plumas County, California, collecting Mecoptera for DNA retrieval to help elucidate the phylogeny of the group. After collecting his targeted species, he suffered a severe …

Using Publicly Available Data To Quantify Plant–Pollinator Interactions And Evaluate Conservation Seeding Mixes In The Northern Great Plains, Clint R.V. Otto, S. O'Dell, R. B. Bryant, N. H. Euliss Jr., R. M. Bush, Matthew D. Smart

Department of Entomology: Faculty Publications

Concern over declining pollinators has led to multiple conservation initiatives for improving forage for bees in agroecosystems. Using data available through the Pollinator Library (npwrc.usgs.gov/pollinator/), we summarize plant–pollinator interaction data collected from 2012–2015 on lands managed by the U.S. Fish and Wildlife Service and private lands enrolled in U.S. Department of Agriculture conservation programs in eastern North Dakota (ND). Furthermore, we demonstrate how plant–pollinator interaction data from the Pollinator Library and seed cost information can be used to evaluate hypothetical seeding mixes for pollinator habitat enhancements. We summarize records of 314 wild bee and 849 honey bee (Apis mellifera …

Thiamethoxam Toxicity And Effects On Consumption Behavior In Orius Insidiosus (Hemiptera: Anthocoridae) On Soybean, Carolina Camargo, Thomas Hunt, Loren J. Giesler, Blair D. Siegfried

Department of Entomology: Faculty Publications

Neonicotinoid residues can be present in soybean vegetative tissue, prey insects, and flower tissues, possibly making them toxic to pollinators and natural enemies. Baseline information on the toxicity of neonicotinoids to beneficial insects other than pollinators through multiple routes of insecticide exposure is limited. The objectives of this study were 1) to evaluate the toxicity of thiamethoxam to the hemipteran predator, Orius insidiosus Say, exposed to residues through treated vegetative tissue and insect prey, and 2) to evaluate the effect of thiamethoxam on the abundance of this predator species in soybean fields. Predators were exposed to thiamethoxam in soybean leaves …

Atp-Sensitive Inwardly Rectifying Potassium Channel Modulators Alter Cardiac Function In Honey Bees, Scott T. O'Neal, Daniel R. Swale, Jeffrey R. Bloomquist, Troy D. Anderson

Department of Entomology: Faculty Publications

ATP-sensitive inwardly rectifying potassium (KATP) channels couple cellular metabolism to the membrane potential of the cell and play an important role in a variety of tissue types, including the insect dorsal vessel, making them a subject of interest not only for understanding invertebrate physiology, but also as a potential target for novel insecticides. Most of what is known about these ion channels is the result of work performed in mammalian systems, with insect studies being limited to only a few species and physiological systems. The goal of this study was to investigate the role that K_ATP channels play in …

Atp-Sensitive Inwardly Rectifying Potassium Channel Regulation Of Viral Infections In Honey Bees, Scott T. O'Neal, Daniel R. Swale, Troy D. Anderson

Department of Entomology: Faculty Publications

Honey bees are economically important pollinators of a wide variety of crops that have attracted the attention of both researchers and the public alike due to unusual declines in the numbers of managed colonies in some parts of the world. Viral infections are thought to be a significant factor contributing to these declines, but viruses have proven a challenging pathogen to study in a bee model and interactions between viruses and the bee antiviral immune response remain poorly understood. In the work described here, we have demonstrated the use of flock house virus (FHV) as a model system for virus …

Weight Of Evidence Evaluation Of A Network Of Adverse Outcome Pathways Linking Activation Of The Nicotinic Acetylcholine Receptor In Honey Bees To Colony Death, Carlie A. Lalone, Daniel L. Villeneuve, Judy Wu-Smart, Rebecca Y. Milsk, Keith Sappington, Kristina V. Garber, Justin Housenger, Gerald T. Ankley

Department of Entomology: Faculty Publications

Ongoing honey bee (Apis mellifera) colony losses are of significant international concern because of the essential role these insects play in pollinating crops. Both chemical and non-chemical stressors have been implicated as possible contributors to colony failure; however, the potential role(s) of commonly-used neonicotinoid insecticides has emerged as particularly concerning. Neonicotinoids act on the nicotinic acetylcholine receptors (nAChRs) in the central nervous system to eliminate pest insects. However, mounting evidence indicates that neonicotinoids also may adversely affect beneficial pollinators, such as the honey bee, via impairments on learning and memory, and ultimately foraging success. The specific mechanisms linking …

Agronomic Potential Of Using Precipitated Calcium Carbonate On Early Plant Growth And Soil Quality In The Intermountain West - Greenhouse Studies, Gary W. Hergert, Murali K. Darapuneni, Abdullah M. Aqeel, Robert G. Wilson, Robert M. Harveson, Jeff D. Bradshaw, Rex A. Nielsen

Department of Entomology: Faculty Publications

Storage and management of large piles of precipitated calcium carbonate (PCC) from sugarbeet processing are a challenge in the western US. Potential uses of this product on surrounding agricultural lands in western NE, eastern WY and northeast CO requires an evaluation of chemical and agronomic impacts of PCC on soils and crop growth. A preliminary greenhouse study was conducted in Scottsbluff, NE using 10 soils from the 3 states. Soils were mixed with 11, 22, 33 and 44 Mg ha^-1 rates of PCC to test the early plant growth of sugarbeet, corn, and dry bean in addition to determining …

Wheat Streak Mosaic Virus Coat Protein Is A Determinant For Vector Transmission By The Wheat Curl Mite, Satyanarayana Tatineni, Anthony J. Mcmechan, Gary L. Hein

Department of Entomology: Faculty Publications

Wheat streak mosaic virus (WSMV; genus Tritimovirus; family Potyviridae), is transmitted by the wheat curl mite (Aceria tosichella Keifer). The requirement of coat protein (CP) for WSMV transmission by the wheat curl mite was examined using a series of viable deletion and point mutations. Mite transmission of WSMV was completely abolished with deletions comprising CP amino acids 58–100. In contrast, the amino-proximal (amino acids 6–27 and 36–57) and carboxy-terminal (14 amino acids) regions of CP were expendable for mite transmission. Mutation of aspartic acid residues at amino acid positions 289 or 326 (D289A or D326A) at the …

Acute-Contact And Chronic-Systemic In Vivo Bioassays: Regional Monitoring Of Susceptibility To Thiamethoxam In Soybean Aphid (Hemiptera: Aphididae) Populations From The North Central United States, Matheus G.P.M. Ribeiro, Thomas E. Hunt, Blair Siegfried

Department of Entomology: Faculty Publications

The risks associated with soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), in the North Central soybean systems has fostered the adoption of prophylactic chemical control practices, such as planting seeds treated with neonicotinoid insecticides, especially thiamethoxam. Consequently, there is a concern that increased selection pressure imposed on the arthropod–pest complex by this insecticide may lead to resistance. Therefore, in vivo bioassays were conducted to determine the susceptibility of soybean aphid to thiamethoxam among North Central U.S. populations. Concentration-mortality data were collected using contact glass-vial and detached-leaf systemic bioassays. The results of these experiments indicate that both bioassays were reliable to …

A New Species Of Golofa Hope, 1837 (Coleoptera: Scarabaeidae: Dynastinae) From Peru, Brett C. Ratcliffe, Stéphane Le Tirant

Department of Entomology: Faculty Publications

A new species of Golofa Hope is described from Peru along with supporting illustrations and a diagnosis.

Se describe una nueva especie de Golofa Hope de Peru soportado por ilustraciones y diagnosis.

Sublethal Effects Of Vatpase-A And Snf7 Dsrnas On Biology Of Southern Corn Rootworm, Diabrotica Undecimpunctata Howardi Barber, Adriano E. Pereira, Ana María Vélez, Lance J. Meinke, Blair D. Siegfried

Department of Entomology: Faculty Publications

RNA interference is a powerful tool against corn rootworm. Adults and neonates of southern corn rootworm, Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae), were exposed to the LC₅₀ of vATPase-A and Snf7 double-stranded RNAs (dsRNAs), and the effects on female fecundity, egg viability, male fitness as measured by sperm viability and mating capacity, larval recovery along with dry weight, and instar determination 10 d after exposure to dsRNA, were determined. Significant reductions were observed for a number of parameters in dsRNA-exposed rootworms relative to control treatments. Female fecundity and larval recovery were significantly reduced after exposure to both dsRNAs. In …

Evaluation Of Smartstax And Smartstax Pro Maize Against Western Corn Rootworm And Northern Corn Rootworm: Efficacy And Resistance Management, Graham P. Head, Matthew W. Carroll, Sean P. Evans, Dwain M. Rule, Alan R. Willse, Thomas L. Clark, Nicholas P. Storer, Ronald D. Flannagan, Luke W. Samuel, Lance Meinke

Department of Entomology: Faculty Publications

Background: Cases of western corn rootworm (WCR) field-evolved resistance to Cry3Bb1 and other corn rootworm (CRW) control traits have been reported. Pyramid products expressing multiple CRW traits can delay resistance compared to single trait products. We used field studies to assess the pyramid CRW corn products, SmartStax (expressing Cry3Bb1 and Cry34Ab1/Cry35Ab1) and SmartStax PRO (expressing Cry3Bb1, Cry34Ab1/Cry35Ab1 and DvSnf7), at locations with high WCR densities and possible Cry3Bb1 resistance, and to assess the reduction in adult emergence attributable to DvSnf7 and other traits. Insect resistance models were used to assess durability of SmartStax and SmartStax PRO to WCR resistance.

Results: …

Identification, Biology, Impacts, And Management Of Stink Bugs (Hemiptera: Heteroptera: Pentatomidae) Of Soybean And Corn In The Midwestern United States, Robert L. Koch, Daniela T. Pezzini, Andrew P. Michel, Thomas Hunt

Department of Entomology: Faculty Publications

Stink bugs (Hemiptera: Heteroptera: Pentatomidae) are an emerging threat to soybean and corn production in the midwestern United States. An invasive species, the brown marmorated stink bug, Halyomorpha halys (Sta° l), is spreading through the region. However, little is known about the complex of stink bug species associated with corn and soybean in the midwestern United States. In this region, particularly in the more northern states, stink bugs have historically caused only infrequent impacts to these crops. To prepare growers and agricultural professionals to contend with this new threat, we provide a review of stink bugs associated with soybean and …

Evaluation Of Foliar Insecticides For The Control Of Western Bean Cutworm In Field Corn, 2015, Débora G. Montezano, Kayla A. Mollet, Grace E. Hirzel, Julie A. Peterson

Department of Entomology: Faculty Publications

The western bean cutworm (WBC) is an important pest of corn and dry beans. This study was conducted within the historic range of WBC in western Nebraska; however, it has undergone a rapid range expansion into the eastern Corn Belt within the last 16 years. This field trial was established to evaluate the efficacy of a single application of foliar insecticides against this pest to prevent feeding damage to non-Bt corn ears. The trial was located at the University of Nebraska-Lincoln’s West Central Water Resources Field Laboratory in Keith County, Nebraska, USA (41.160246° N, –102.035695° W). A RCB design with …

Evaluation Of Foliar Insecticides For The Control Of Western Bean Cutworm In Field Corn, 2016, Débora G. Montezano, Kayla A. Mollet, Grace E. Hirzel, Julie A. Peterson

Department of Entomology: Faculty Publications

The western bean cutworm (WBC) is an important pest of corn and dry beans. In addition to yield loss due to direct feeding on developing kernels in the ear, WBC infestation can also lead to secondary fungal infections. This study was conducted within the historic range of WBC in western Nebraska; however, it has undergone a rapid range expansion into the eastern Corn Belt within the last 16 years. This field trial was established to evaluate the efficacy of a single application of foliar insecticides against this pest to prevent feeding damage to non-Bt corn ears. The trial was located …

Rainbow Peacock Spiders Inspire Miniature Superiridescent Optics, Bor-Kai Hsiung, Radwanul Hasan Siddique, Doekele G. Stavenga, Jürgen C. Otto, Michael C. Allen, Ying Liu, Yongfeng Lu, Dimitri D. Deheyn, Matthew D. Shawkey, Todd A. Blackledge

Department of Electrical and Computer Engineering: Faculty Publications

Colour produced by wavelength-dependent light scattering is a key component of visual communication in nature and acts particularly strongly in visual signalling by structurallycoloured animals during courtship. Two miniature peacock spiders (Maratus robinsoni and M. chrysomelas) court females using tiny structured scales (~ 40 × 10 μm²) that reflect the full visual spectrum. Using TEM and optical modelling, we show that the spiders’ scales have 2D nanogratings on microscale 3D convex surfaces with at least twice the resolving power of a conventional 2D diffraction grating of the same period. Whereas the long optical path lengths required …

Importance Of The Antenniform Legs, But Not Vision, For Homing By The Neotropical Whip Spider Paraphrynus Laevifrons, Verner P. Bingman, Jacob M. Graving, Eileen Hebets, Daniel D. Wiegmann

Eileen Hebets Publications

Amblypygids, or whip spiders, are nocturnal, predatory arthropods that display a robust ability to navigate to their home refuge. Prior field observations and displacement studies in amblypygids demonstrated an ability to home from distances as far away as 10 m. In the current study, micro-transmitters were used to take morning position fixes of individual Paraphrynus laevifrons following an experimental displacement of 10 m from their home refuge. The intention was to assess the relative importance of vision compared with sensory input acquired from the antenniform legs for navigation as well as other aspects of their spatial behavior. Displaced individuals were …

Phytophagous Insect Oviposition Shifts In Response To Probability Of Flower Abortion Owing To The Presence Of Basal Fruits, Shivani Jadeja, Brigitte Tenhumberg

School of Biological Sciences: Faculty Publications

Phytophagous insects use a wide range of indicators or associated cues to avoid laying eggs in sites where offspring survival is low. For insects that lay eggs in flowers, these unsuitable sites may be created by the host plant’s resource allocation to flowers. In the sequentially flowering host plant, Yucca glauca, late-opening distal flowers are more likely to be aborted in the presence of already-initiated basal fruits because they are strong resource sinks. If flowers are aborted, all eggs of the phytophagous insect, Tegeticula yuccasella, within the flower die. We used the phytophagous insect T. yuccasella that lays …

Distinct Fitness Costs Associated With The Knockdown Of Rnai Pathway Genes In Western Corn Rootworm Adults, Ke Wu, Carolina Camargo, Elane Fishilevich, Kenneth E. Narva, Xiuping Chen, Caitlin E. Taylor, Blair D. Siegfried

Department of Entomology: Faculty Publications

RNA interference (RNAi) based approaches can potentially be used to control insect pests. These approaches may depend on the usage of microRNA (miRNA) or double stranded RNA (dsRNA) mediated gene knockdown, which likely involves proteins that regulate these pathways, such as Argonaute 1 (Ago1), Argonaute 2 (Ago2), Dicer 1 (Dcr1), Dicer 2 (Dcr2), and Drosha in insects. We previously performed functional characterization of Ago2 and Dcr2 of western corn rootworm (WCR), Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) and observed that knockdown of Ago2 and Dcr2 ameliorated the lethal effect induced by the dsRNA-mediated knockdown of an essential gene in WCR, thereby …

“Hummingbird” Floral Traits Interact Synergistically To Discourage Visitation By Bumble Bee Foragers, Robert J. Gegear, Rebecca Burns, Katharine A. Swoboda-Bhattarai

Department of Entomology: Faculty Publications

Pollination syndromes are suites of floral traits presumed to reflect adaptations to attract and utilize a “primary” type of animal pollinator. However, syndrome traits may also function to deter “secondary” flower visitors that reduce plant fitness through their foraging activities. Here we use the hummingbird-pollinated plant species Mimulus cardinalis as a model to investigate the potential deterrent effects of classic bird syndrome traits on bumble bee foragers. To establish that M. cardinalis flowers elicit an avoidance response in bees, we assessed the choice behavior of individual foragers on a mixed experimental array of M. cardinalis and its bee-pollinated sister species …

Examination Of Civil Forensic Entomology Through Case Studies, Alicia Thuis

Department of Entomology: Distance Master of Science Projects

Brief History of Forensic Entomology 2 Civil Forensic Case Studies 11 The Raisin Ant 11 Maggots in My Potato! 13 Extra Protein for Fido 15 Health Food Invasion 16 Swimming Spiders 18 Extras in the Steak 19 In or Out? 23 Buttery Parasites 24 Laminated Flies 26 Chicken Fried Maggots 27 Process of Elimination 29 Bed Bugs Everywhere! 30 Not a “Bug” 32 Not a Cockroach 34 Ethical Considerations 36 Best Practice Recommendations 43 Future Research Directions 47 References 50

Biology And Management Of Key Rice Pest In Asia, Airene Millsap

Department of Entomology: Distance Master of Science Projects

1. Importance of rice

2. Origin and history of rice as a crop

3. Rice cultivation 3.1 Lowland or Irrigated rice 3.2 Upland rice 3.3 Deep water rice 3.4 Floating rice

4. Parts of rice plants 4.1 Vegetative stage 4.2 Reproductive stage 4.3 Ripening stage

5. Rice Pests

5.1 Vegetative stage • Stem Borer • Gall Midge • Rice Hispa • Rice Leaf Folder • Rice Armyworms • Rice Whorl Maggots • Rice Caseworm • Rice Mealybugs • Short Horned Grasshopper • Rice Stem Maggot • Rice Thrips • Oriental Migratory Locust • Black Bug • Green Stink Bug

5.2 …

Invasive Species Impacts On Infrastructure, Invasive Species Advisory Committee

National Invasive Species Council

Executive Summary

Invasive species represent one of the most significant threats to ecosystems, human and animal health, infrastructure, the economy, and cultural resources. Because potentially invasive, non-native species typically enter the United States through ports of entry in urban environments, some of the first observable impacts may be to infrastructure, yet little is known about the economic costs associated with these impacts to the “built” environment. In addition, federal agencies currently lack the authority necessary to effectively prevent, eradicate, and control invasive species that impact the human-built environment. This lack of authority prevents rapid response to some of the most …

Insecticide Sensitivity Of Native Chloride And Sodium Channels In A Mosquito Cell Line, Lacey J. Jenson, Troy D. Anderson, Jeffrey R. Bloomquist

Department of Entomology: Faculty Publications

The aim of this study was to investigate the utility of cultured Anopheles gambiae Sua1B cells for insecticide screening applications without genetic engineering or other treatments. Sua1B cells were exposed to the known insecticidal compounds lindane and DIDS, which inhibited cell growth at micromolar concentrations. In patch clamp studies, DIDS produced partial inhibition (69%) of chloride current amplitudes, and an IC50 of 5.1 μM was determined for Sua1B cells. A sub-set of chloride currents showed no response to DIDS; however, inhibition (64%) of these currents was achieved using a low chloride saline solution, confirming their identity as chloride channels. In …

Resistance Of Rice Varieties To Sitophilus Oryzae (Coleoptera: Curculionidae), Diana Cristina Da Silva Costa, André Cirilo De Sousa Almeida, Marcio Da Silva Araújo, Elvis Arden Heinrichs, Mabio Chrisley Lacerda, José Alexandre Freitas Barrigossi, Flávio Gonçalves De Jesus

Department of Entomology: Faculty Publications

Abstract

Rice, Oryza sativa L. (Poaceae), is one of the world’s most important food crops. Among the insects that damage rice grains, the rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), is the most important species. The objective of this study was to identify rice varieties with resistance to S. oryzae as measured by grain feeding damage and S. oryzae reproduction. The rice varieties evaluated were obtained from the germplasm bank of the National Center of Research Rice and Beans, Embrapa, Brazil. The experimental design was completely randomized with 3 replications and 22 treatments (varieties). The varieties Bonança, Esmeralda, and Rio …

Integration Of Plant Defense Traits With Biological Control Of Arthropod Pests: Challenges And Opportunities, Julie A. Peterson, Paul J. Ode, Camila Oliveira-Hofman, James D. Harwood

Department of Entomology: Faculty Publications

Crop plants exhibit a wide diversity of defensive traits and strategies to protect themselves from damage by herbivorous pests and disease. These defensive traits may be naturally occurring or artificially selected through crop breeding, including introduction via genetic engineering. While these traits can have obvious and direct impacts on herbivorous pests, many have profound effects on higher trophic levels, including the natural enemies of herbivores. Multi-trophic effects of host plant resistance have the potential to influence, both positively and negatively, biological control. Plant defense traits can influence both the numerical and functional responses of natural enemies; these interactions can be …