The Larval Stage and Life Cycle of Diplodontus scapularis Dugès, 1834, and D. semiperforatus Walter, 1925 (Acari: Hydrachnidia), with Remarks on Their Habitat Preference and Distribution in the South West Palaearctic Region

The larva of D. scapularis Dugès, 1834, is redescribed from material collected in Morocco, and a comparison is made with larvae of the closely related D. semiperforatus (Walter, 1925) reared in the laboratory. Larvae of D. scapularis were found parasitic in the subelytral space of several beetle species of the families Hydraenidae, Hydrochidae and Hydrophilidae. New records are given of Diplodontus scapularis from Morocco and of D. semiperforatus from Spain. The two species differ in salinity tolerance, with D. semiperforatus obviously adapted to saline and hypersaline inland waters.     

Influence of toxic and non-toxic phytoplankton on feeding and survival of Dreissena polymorpha (Pallas) larvae

Grazing and survival of larvae of the zebra mussel, Dreissena polymorpha, on a green alga and cyanobacteria were studied in laboratory experiments. Clearance rates of the larvae were determined for Chlamydomonas reinhardtii (green alga), two non-toxic and two toxic Microcystis aeruginosa strains (Cyanobacteria). Clearance rates of larvae on non-toxic Microcystis were significantly higher than on toxic Microcystis. The clearance rate on Chlamydomonas reinhardtii was in between the clearance rates on toxic and non-toxic Microcystis strains and not significantly different from them. Effects of toxicity of Microcystis on the survival of zebra mussel larvae was investigated in a short-term experiment. Survival of larvae fed toxic Microcystis was lower than that of larvae fed non-toxic Microcystis, but higher than that of starved larvae. This may imply that, for survival of zebra mussel larvae, it is better to have bad quality (toxic) food than no food.     

Effects of insect damage on photosynthesis,transpiration and SO2 uptake by sycamore

Summary The bilberry (Vaccinium myrtillus L.), is the dominant plant in the field layer of many boreal forests in northern Sweden. It is utilized by several herbivorous insect larvae as food (i.e. Lepidoptera: Geometridae, Tortricidae and Hymenoptera: Symphyta). Total density of insect larvae was 63% lower where birds had access to larvae compared to exclosures. Larvae with a feeding behaviour which exposes them to birds (geometrids and sawflies) showed a pronounced reduction in density when exposed to bird predators. Density of larvae with a concealed feeding behaviour (tortricids) was unaffected by birds. The frequency of larval damage to bilberry annual shoots was significantly higher in areas where larval density was high due to exclusion of birds. I concluded that birds have the potential to heavily modify the interaction between bilberry and insect larvae.     

Insectivory and social digestion inDrosophila

It has long been believed thatDrosophila larvae feed almost entirely by ingesting yeast and possibly other microorganisms that are associated with fermenting fruits or other vegetable matter. However, we have discovered that the larvae of a number ofDrosophila species can consume such diverse substrates as insect tissues, including the exoskeleton. Experiments reported here, which include raising sterile dechorionated eggs to adulthood on adult carcasses under axenic conditions, show that larvae can consume complex chitinous substrates directly without the assistance of microorganisms. We show thatDrosophila larvae are able externally to digest amylose, cellulose, and chitin, without coming into physical contact with them. We conclude that not only doDrosophila larvae produce enzymes enabling them to digest a wide variety of substrates, but also these enzymes are egested onto the substrates so that at least some digestion, especially of large polymers, takes place externally. Finally, we suggest that the phenomenon of external digestion explains both the previously unexplained massiveness ofDrosophila salivary glands and their chromosomes and the tendency of larvae to cluster, which may also be true of other dipterans.This investigation was supported by NIH (AREA) Grant IR 15AG05897.     

The free swimming Pipa larvae,with a review of pipid larvae and pipid phylogeny (Anura: Pipidae)

This paper describes the morphology of the free swimming Pipa larvae, compares them with Xenopus, Hymenochirus, and to some extent, Rhinophrynus larvae, and presents a morphological diagnosis of pipid larvae. Pipa and Xenopus have very similar chondrocrania. Hymenochirus is superficially different but has the same diagnostic features. The differences appear related to its small size and predatory habitus. Other aspects of anatomy, especially the filter apparatus are very different in each genus. The filter apparatus of Pipa is somewhat reduced and seems modified for the retention of relatively large (20+microns) particles. Similar adaptations may have been annectant to predations in Hymenochirus, which lacks a filter apparatus. However, varying states of seven character complexes, which cut across the varying ecology, show that there are two basic pipid lineages, each currently confined to Africa or South America, respectively. Recent finds of fossil South American Xenopus indicate that these two lineages separated before the continents did. This does not warrant the recognition of two subfamilies because Xenopus and Hymenochirus are too different. Pseudhymenochirus is not an intermediate between them; it is a primitive Hymenochirus. Eight character states separate pipid and rhynophrynid larvae.     

Phylogeny of Hydrophiloidea (Coleoptera: Staphyliniformia) using characters from adult and preimaginal stages

A phylogenetic analysis using characters from egg cases, larvae, pupae and adults was conducted; the outgroups included the beetle families Silphidae, Hydraenidae and Histeridae. Characters from the immature stages were obtained mostly from material reared in the laboratory, those from the adults were obtained from Hansen's generic revision for the superfamily. The results support the position of Hydraenidae within the Staphylinoidea, and not as part of Hydrophiloidea; Histeroidea is proposed as the sister group of Hydrophiloidea. At family level two clades are distinguished; the relationships within the first clade are ((Georissidae Epimetopidae) Helophoridae), those within the second are ((Hydrophilidae Spercheidae) Hydrochidae). Larval characters were most informative at the base of the tree, especially those associated with the spiracular atrium; adult characters were most informative at the apex of the tree.     

Effects of spatio-temporal intervals between newly-hatched larvae on larval survival and development inMonochamus alternatus (Coleoptera: Cerambycidae)

The effects of distance between hatching larvae on survival and development were investigated inMonochamus alternatus. Two newly-hatched larvae were inoculated intoPinus densiflora bolts at a distance of 2.5 cm or 10 cm, simultaneously or at an interval of 2 weeks. Some larvae were inoculated singly as a control. When larvae were inoculated simultaneously, mortality of the closely-inoculated larvae was significantly higher than that of distantly-inoculated larvae. Such high mortality was identified as due to conspecific bites. When the two larvae were inoculated asynchronously, the first-inoculated larvae killed some second-inoculated larvae but were never killed by them. Consequently, mortality was higher in second-inoculated larvae than in first-inoculated larvae. In particular, there was a significant difference in mortality between them when the larvae had been inoculated closely. The mortality of second-inoculated larvae was higher in the closely-inoculated group than in the distantly-inoculated group although there was no significant difference between them. In the case of two simultaneously-inoculated larvae, the initial distance between them had no significant effect on the development and growth in the early larval stage. When the larvae were inoculated asynchronously, the first-inoculated larvae grew more quickly than singly-inoculated control larvae.     

Absence of cuticular alkenes allows lycaenid larvae to avoid predation by Formica japonica ants

Chemical mimicry and camouflage based on cuticular hydrocarbons (CHCs) are adaptive strategies that are frequently observed in myrmecophilous insects. The larvae of several lycaenid butterfly species that exhibit obligate associations with specific ant species have been reported to use chemical mimicry. However, little is known about the strategies used by the larvae of species that have facultative associations with multiple ant species. We attempted to reveal the effects of larval CHC profiles on interactions with Formica japonica workers, using three lycaenid species, two facultative ant‐associated (Lycaeides argyrognomon and Zizeeria maha) and one non‐ant‐associated (Lycaena phlaeas), which commonly possess n‐alkanes as the major CHCs. In field bioassays, the lycaenid larvae were attacked by ant workers less often than larvae of Papilio polytes (Papilionidae), the CHCs of which were rich in 7‐alkenes. Treating the lycaenid larvae with 7‐heptacosene and 9‐heptacosene significantly activated ant aggression (biting), whereas treating them with n‐heptacosane, n‐octacosane and 13‐methylheptacosane had little effect. Furthermore, larvae of Pieris rapae (Pieridae), possessing n‐alkanes as the dominant CHCs, suffered an intermediate level of ant biting between the lycaenid and Pa. polytes larvae. However, treatments of the P. rapae larvae with 7‐heptacosene and 9‐heptacosene significantly affected the frequency of ant biting. These findings suggest that the absence of alkenes in larval CHC profiles is an effective means of circumventing predation by ants and allows lycaenid larvae to inhabit the foraging territory of predaceous ants, at least to some extent.     

Predicting larval metamorphosis of Pacific lamprey Entosphenus tridentatus through measurements of length and mass

The utility of length and mass measurements to predict the larval metamorphosis of Pacific lamprey Entosphenus tridentatus was evaluated. During 2004–2008, larval E. tridentatus were collected from Cedar Creek (Washington, USA) in either the spring or autumn, measured for total length and total mass, reared in captivity and monitored for metamorphosis. The minimum total length, total mass and condition factor of larvae that were observed to go through metamorphosis were 102 mm, 2.0 g and 1.52, respectively. Logistic models indicated that total length and condition factor in both spring and autumn were the most significant variables for predicting metamorphosis of Pacific lamprey during the subsequent summer. Mass in the autumn also appeared important to predict whether metamorphosis occurred in the subsequent summer. Collectively, all models using specific minimums of total length, total mass or condition factor of larvae as criteria for them to metamorphose were sometimes (5 of 14 cases) able to predict the percentage of larvae that would metamorphose but rarely (1 of 12 cases) able to predict which individual larvae would metamorphose. Similar to other anadromous species of lampreys, the size and condition of larval E. tridentatus have utility for predicting metamorphic fate.     

A field study of potential ecological costs of resistance by ‘stem ducking’ in tall goldenrod,Solidago altissima

Culex quinquefasciatus Say (Diptera: Culicidae) is an abundant urban mosquito that is the vector of filariasis. Breeding in septic tanks, where there are very high levels of bacterial food, it is likely to have a different reaction to crowding compared with other mosquitoes. To test for the presence and type of crowding effects, four larval densities of C. quinquefasciatus varying from 0.4 to 3.2 larvae ml^?1 of water were reared in tubes. Mortality was found to greatly increase at densities above 0.8 larvae, whereas larval duration increased even above 0.4 larvae ml^?1. Changing the water in the tubes daily gave a small (but significant) response in reducing mortality and larval duration. However, when larvae kept at a low density shared the same water with larvae at high density, there was no chemical influence on their growth rate and mortality. The effect of crowding was primarily due to physical disturbances between larvae. When larvae were kept at a high density in the same volume of water, but in shallow trays with a large surface area and therefore much less contact between them, mortality was the same as for the lowest density. There was still, however, a significant increase in larval duration from 8.6 days at 0.4 larvae ml^?1 to 12.1 days at 3.2 larvae ml^?1. It is therefore concluded that the larvae respond to physical rather than chemical factors by prolonging larval development and having some increase in mortality.     

Dietary plant phenolic improves survival of bacterial infection in Manduca sexta caterpillars

Plant phenolics are generally thought to play significant roles in plant defense against herbivores and pathogens. Many plant taxa, including Solanaceae, are rich in phenolic compounds and some insect herbivores have been shown to acquire phenolics from their hosts to use them as protection against their natural enemies. Here, we demonstrate that larvae of an insect specialist on Solanaceae, the tobacco hornworm, Manduca sexta L. (Lepidoptera: Sphingidae), acquire the plant phenolic chlorogenic acid (CA), and other caffeic acid derivatives as they feed on one of their hosts, Nicotiana attenuata L. (Solanaceae), and on artificial diet supplemented with CA. We test the hypothesis that larvae fed on CA‐supplemented diet would have better resistance against bacterial infection than larvae fed on a standard CA‐free diet by injecting bacteria into the hemocoel of fourth instars. Larvae fed CA‐supplemented diet show significantly higher survival of infection with Enterococcus faecalis (Andrewes & Horder) Schleifer & Kilpper‐Bälz, but not of infection with the more virulent Pseudomonas aeruginosa (Schroeter) Migula. Larvae fed on CA‐supplemented diet possess a constitutively higher number of circulating hemocytes than larvae fed on the standard diet, but we found no other evidence of increased immune system activity, nor were larvae fed on CA‐supplemented diet better able to suppress bacterial proliferation early in the infection. Thus, our data suggest an additional defensive function of CA to the direct toxic inhibition of pathogen proliferation in the gut.     

Moving on hairy surfaces: modifications of Gratiana spadicea larval legs to attach on its host plant Solanum sisymbriifolium

Both larvae and adults of Gratiana spadicea (Klug) (Coleoptera: Chrysomelidae: Cassidinae) feed exclusively on leaves of Solanum sisymbriifolium Lamarck (Solanaceae), which have simple and stellate trichomes. The simple trichomes can be non-glandular or glandular. The stellate trichomes present a long central ray that secretes a viscous exudate when broken. Trichome effects on movement of G. spadicea larvae were evaluated in the laboratory. Larval speed on intact petioles of S. sisymbriifolium was compared to those where the exudates and/or stellate trichomes were removed. Exudates had no effect on larval speed. Stellate trichomes mechanically slowed down first instar movement. Gratiana spadicea larval legs have a modified distal portion, the tarsungulus, whose rounded aperture shape matches that of the cylindrical pointed rays of S. sisymbriifolium stellate trichomes. The first three instars anchor the tarsungulus to the trichome rays, and get both attachment to the leaf surface and support to body impulsion. Morphological comparisons showed that the legs in the first larval instars are shorter than the central ray of the stellate trichome, so larvae have to walk above them. Fifth instar larvae have longer legs and walk by inserting the sharp tip of their tarsungulus directly into the leaf epidermis. The dimensions of the tarsungulus aperture vary in such a way that it can clasp every thickness of trichome ray for any larval instar. Contrary to other tarsungulus portions and larval body features, where growth is allometric, growth of the tarsungulus aperture is isometric throughout the larval stage. Thus, it is suggested that this G. spadicea leg structure is adapted for moving on hairy leaf surface of S. sisymbriifolium.     

Concurrently burrowing host fly larvae reciprocally enhance pupation depth to minimise parasitism risk

1. Variation in parasitism risk among hosts is a key factor influencing host–parasitoid interactions; however, within-patch variation (as opposed to between-patch variation) in parasitism risk has hardly been studied. This study investigated the mechanisms of within-patch variation in parasitism risk in the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). 2. Bactrocera dorsalis pupates underground. Previous studies have shown that pupae at greater depths have a reduced risk of parasitism by a pupal parasitoid Dirhinus giffardii (Silvestri) (Hymenoptera: Chalcididae). When a pupa near the ground surface is parasitised, the victim may decrease the risk of parasitism risk for pupae located at deeper depths. It was hypothesised that larvae will pupate at greater depths when they perceive the presence of conspecifics because of the benefit of pupating deeper than other individuals. 3. In a series of laboratory experiments, the effects of three factors on pupation depth were examined, including: (i) the density of larvae during development; (ii) the density of concurrently burrowing larvae; and (iii) the presence of pre-existing pupae in the pupation substrate. Only the density of concurrently burrowing larvae influenced pupation depth, which suggests that when a burrowing larva perceives the presence of other burrowing larvae, it aims to burrow deeper than them. 4. This study shows not only that parasitism risk is variable among pupae within a patch, but also that it is density-dependent. A commonly made assumption (i.e. the absence of within-patch variability in parasitism risk) needs to be re-evaluated in a wide range of host–parasitoid systems.     

Intraguild predation, cannibalism, and microhabitat use in Calopteryx virgo and Somatochlora metallica larvae: a laboratory experiment

Intraguild predation (IGP) and cannibalism among co-occurring lotic odonate species was studied in Central Finland. A laboratory experiment was performed to assess the microhabitat use and cannibalism between intermediate and late instars of Calopteryx virgo larvae and predation by larger Somatochlora metallica larvae on the intermediate C. virgo instars. The experiment was run in small running-water aquaria where the larvae were able to divide their mutual habitat vertically by clinging onto artificial perches or crawling on the bottom. Life span of the small C. virgo larvae and attack rate on them were compared between the cannibalism and IGP treatments. The effect of predation on the activity, habitat use and spatial distribution of the small C. virgo larvae was examined. The IGP rate was 36%. The prey larvae spent the most of their time on the perches, whereas the S. metallica preferred the substrate. The large C.␣virgo larvae did not cannibalise smaller conspecifics. The presence of a predator (S. metallica) had no effect on the habitat use or activity of the prey (C. virgo) larvae. Habitat use differed more between those species than between conspecifics of different size classes of C. virgo. The spatial distribution between S.␣metallica and C. virgo showed a completely random pattern, whereas the two size classes of C. virgo aggregated in the vegetation. Absence of cannibalism and behavioural observations indicate that C. virgo may have a low tendency for intraspecific aggressions.     

Sites chosen by diapausing or quiescent stage quino checkerspot butterfly, <Emphasis Type="Italic">Euphydryas editha quino</Emphasis>, (Lepidoptera: Nymphalidae) larvae

This study examines whether in nature endangered quino checkerspot (Euphydryas editha quino) larvae will return to diapause and if so where they choose to hide. Multiple years of diapause probably help larvae survive drought years and sites chosen have high survival value to the species. Ninety square meters of habitat were created by removing non native plants and replacing them with natives found at checkerspot occupied sites. During the 2005–2006 winter 1,000 post-diapause larvae were released. From these larvae 31 adults (20 males and 11 females) developed over a 2.5 month period (March 20–June 6) from 41 pupae. One chrysalis was parasitized by a parasitic wasp Pteromalus puparum (L.) in the family Pteromalidae, one was partially eaten by an animal, while the remaining eight pupae died of unknown causes. Thirty quadrats (1 square meter each) were cleared of vegetation, leaf and branch litter, rocks, and checkerspot larvae from July 5 to August 1, 2006. Forty-nine larvae were found that returned to diapause. Most larvae (31) chose to make shelters on California buckwheat, which is not a checkerspot food plant, two to five cm above the ground. One shelter had 22, another had seven, and two others had single larvae. Five of 10 larvae found in leaf litter below California buckwheat were crawling and not associated with shelters suggesting they had been dislodged from shelters. California buckwheat may be important in habitat restoration for the checkerspot, particularly at sites below 900 meters elevation where summer conditions are hot and dry. No additional larvae were found the following spring, when they should have exited diapause. Therefore 910 (91%) larvae were lost to some undocumented form of mortality.     

Biology of epiphytic Chironomidae (Diptera:Nematocera) in chalk streams

Submerged macrophytes are a conspicuous feature of chalk stream ecosystems, supporting large populations of diatoms and invertebrates, including larvae of the nematocerous family Chironomidae. Close temporal and spatial association is evident between chironomid larvae and diatoms, and diatoms are frequently a major component of the food of midge larvae. Larvae provided with food that is rich in diatoms survive better and grow and develop more quickly than larvae that are supplied with food that is predominantly fine organic detritus. First instar larvae selectively feed on diatoms but it is likely that selection is on the basis of particle size, rather than for diatoms per se. Three species of epiphytic larvae (2 Cricotopus spp. and Eukiefferiella ilkleyensis) transfer to a diet that is predominantly of fresh plant material (Ranunculus calcareus leaves) in the third and fourth instar. The life cycle strategies of many species of Orthocladiinae equip them to exploit temporarily favourable environmental conditions very rapidly and effectively. This fact helps to explain the coexistence of species with similar strategies of resource utilization, as part of the normal epiphytic fauna of chalk streams.     

Indirect effects of the fairy shrimp,Branchipus schaefferi and two ostracod species on Bacillus thuringiensis var Israelensis-induced mortality in mosquito larvae

Bacillus thuringiensis var israelensis (BTI), when ingested by mosquito larvae, is highly toxic to them. Many other aquatic invertebrates feed on bacteria but, in general, BTI is not toxic to them. We tested in the laboratory the hypothesis that certain crustaceans indirectly benefit mosquito larvae by reducing mortality caused by BTI. We presumed the mechanism to be ingestion of the bacteria by the crustaceans resulting in a lower concentration available to the mosquito larvae.Mortality of Aedes aegypti larvae exposed to BTI was reduced in the presence of the fairy shrimp, Branchipus schaefferi and the ostracod, Cypridopsis vidua (only during summer trials and not autumn trials for the latter species) but was not reduced in the presence of the ostracod, Heterocypris incongruens. By contrast, H. incongruens preyed upon infected, though still-moving, larvae. Feeding on the bacteria by the crustaceans may not be an important mechanism; our data indicates that the crustaceans did not reduce BTI in the water. Moreover, B. schaefferi, introduced into water and then removed prior to the introduction of BTI and mosquitoes, also reduced mosquito mortality.The mechanism for the protective effect of B. schaefferi and C. vidua is unknown. However, these results suggest that the abundance of certain organisms co-occurring with mosquito larvae may partially explain why the effective concentration of BTI varies among habitats.     

Host selection by the autoparasitoid Encarsia pergandiella on primary (Bemisia tabaci) and secondary (Eretmocerus mundus) hosts

In autoparasitoids, females are generally primary endoparasitoids of Hemiptera, while males are hyperparasitoids developing in or on conspecific females or other primary parasitoids. Female‐host acceptance can be influenced by extrinsic and/or intrinsic factors. In this paper, we are concerned with intrinsic factors such as nutritional status, mating status, etc. We observed the behavior of Encarsia pergandiella Howard (Hymenoptera: Aphelinidae) females when parasitizing primary (3rd instar larvae of Bemisia tabaci Gennadius [Homoptera: Aleyrodidae]) and secondary hosts (3rd instar larvae and pupae of Eretmocerus mundus Mercet [Hymenoptera: Aphelinidae]) for a period of 1 h. Females had different reproductive (virgin or mated younger) and physiological (fed elder or mated elder) status. Virgin females killed a large number of secondary hosts while investing a long time per host. However, they did not feed upon them. Mated females killed a lower number of secondary hosts and host feeding was observed in both consuming primary and secondary hosts. It was common to observe host examining females of all physiological statues tested repeatedly stinging the same hosts when parasitizing, killing or rejecting them. Fed elder females parasitized more B. tabaci larvae than E. mundus larvae or pupae, while investing less time on the primary host than on the secondary host. They also parasitized more B. tabaci larvae than mated elder females, while investing less time per host. The access of females to honey allowed them to lay more eggs.     

Influence of aphid honeydew on the foraging behaviour of Hippodamia convergens larvae

1. Environmental cues associated with prey are known to increase predator foraging efficiency. Ladybird larvae are major predators of aphids. The sugary excretion of aphids (honeydew) has been proposed to serve as a prey‐associated cue for ladybird larvae. 2. Ladybird larvae are regularly found on the ground moving between plants or after falling off plants. The use of prey‐associated cues would be particularly beneficial for ladybird larvae on the ground in that such cues would help them to decide which plants to climb because aphids are patchily distributed within as well as amongst plants and, as a result, many plants are either not infested with aphids or do not host an aphid species of high nutritional value for ladybird larvae. 3. Laboratory experiments with larvae of Hippodamia convergens Guérin‐Méneville (Coleoptera: Coccinellidae) were carried out to explore whether honeydew accumulated on the ground is used as a foraging cue. The study also investigated whether, if honeydew is a foraging cue, larvae show differential responses to honeydew of high‐quality prey Acyrthosiphon pisum Harris compared with that of low‐quality prey Aphis fabae Scopoli (both: Homoptera: Aphididae). 4. Hippodamia convergens larvae stayed longer in areas containing honeydew but did not engage in longer bouts of searching. Furthermore, larvae did not distinguish between honeydew from high‐ and low‐quality aphid prey.     

Genetic diversity and population structure of a cyprinid fish (Ancherythroculter nigrocauda) in a highly fragmented river

In this study, samples of Ancherythroculter nigrocauda, an endemic fish in the upper Yangtze River, were collected above and below dams in the Longxi River, a tributary of the upper Yangtze River, China, to investigate the genetic impacts of dams. The mitochondrial cytochrome b gene (cyt b) and 13 microsatellite (SSR) loci were used to analyze whether dams have resulted in loss of genetic diversity of the two fragmented populations or caused genetic differentiation between them. The results showed that the haplotype diversity (0.488; 0.486), nucleotide diversity (0.084%; 0.082%) and average expected heterozygosity (0.652; 0.676) of the two populations were all at a low level, and recent bottlenecks were detected. However, there was no genetic differentiation detected by the low genetic differentiation index (F_st, cyt b: ?0.1677, p = 0.99707; SSR: 0.00259, p = 0.81427). Besides, 11 pairs of half‐sibling relationship were found between the two populations indicating that there were individual movements and gene flow between them. This could be the larvae moving from upstream to downstream when water spilled over dams in flooding season. Therefore, our analysis showed that the dams have caused a loss of genetic diversity of the populations of A. nigrocauda in the Longxi River, blocked the active upstream movement but allowing passive downstream drift of larvae.