A new species of Megaglena Peters amp; Edmunds, 1970 (Ephemeroptera: Leptophlebiidae) from Tamil Nadu, India.

Megaglena sivarubani sp. nov. is described based on the nymphs from the Puliyuthu falls of Tamil Nadu, Southern India. The number of species of Megaglena Peters Edmunds is now expanded to three and it is the first record of the genus from the Tamil Nadu part of Southern Western Ghats. Megaglena sivarubani sp. nov. can be distinguished from Indian species M. agasthiya Vasanth, Subramanian Selvakumar by the body size of the nymphs, shape of the labial palp segment III, and by the setation of foretibia. The morphological variations between all the known species of Megaglena are discussed and summarized.

Regulation of metamorphosis in neopteran insects is conserved in the paleopteran Cloeon dipterum (Ephemeroptera).

In the Paleozoic era, more than 400 Ma, a number of insect groups continued molting after forming functional wings. Today, however, flying insects stop molting after metamorphosis when they become fully winged. The only exception is the mayflies (Paleoptera, Ephemeroptera), which molt in the subimago, a flying stage between the nymph and the adult. However, the identity and homology of the subimago still is underexplored. Debate remains regarding whether this stage represents a modified nymph, an adult, or a pupa like that of butterflies. Another relevant question is why mayflies have the subimago stage despite the risk of molting fragile membranous wings. These questions have intrigued numerous authors, but nonetheless, clear answers have not yet been found. By combining morphological studies, hormonal treatments, and molecular analysis in the mayfly Cloeon dipterum, we found answers to these old questions. We observed that treatment with a juvenile hormone analog in the last nymphal instar stimulated the expression of the Kr-h1 gene and reduced that of E93, which suppress and trigger metamorphosis, respectively. The regulation of metamorphosis thus follows the MEKRE93 pathway, as in neopteran insects. Moreover, the treatment prevented the formation of the subimago. These findings suggest that the subimago must be considered an instar of the adult mayfly. We also observed that the forelegs dramatically grow between the last nymphal instar, the subimago, and the adult. This necessary growth spread over the last two stages could explain, at least in part, the adaptive sense of the subimago.

Genomic adaptations to aquatic and aerial life in mayflies and the origin of insect wings.

The evolution of winged insects revolutionized terrestrial ecosystems and led to the largest animal radiation on Earth. However, we still have an incomplete picture of the genomic changes that underlay this diversification. Mayflies, as one of the sister groups of all other winged insects, are key to understanding this radiation. Here, we describe the genome of the mayfly Cloeon dipterum and its gene expression throughout its aquatic and aerial life cycle and specific organs. We discover an expansion of odorant-binding-protein genes, some expressed specifically in breathing gills of aquatic nymphs, suggesting a novel sensory role for this organ. In contrast, flying adults use an enlarged opsin set in a sexually dimorphic manner, with some expressed only in males. Finally, we identify a set of wing-associated genes deeply conserved in the pterygote insects and find transcriptomic similarities between gills and wings, suggesting a common genetic program. Globally, this comprehensive genomic and transcriptomic study uncovers the genetic basis of key evolutionary adaptations in mayflies and winged insects.

Acute and Chronic Toxicity of Nickel and Zinc to a Laboratory Cultured Mayfly (Neocloeon triangulifer) in Aqueous but Fed Exposures.

Aquatic insects are poorly represented in water quality criteria, and previous studies have suggested a lack of sensitivity in acute toxicity tests despite observational studies demonstrating the contrary. Our objectives were to determine the toxicity of nickel (Ni) and zinc (Zn) to the mayfly Neocloeon triangulifer in fed acute (96-h) and chronic exposures to estimate aqueous effect concentrations while acknowledging the importance of dietary exposure for these insects. For the chronic tests, we conducted preliminary full-life cycle (~25-30 d) and subchronic (14 d) exposures to compare the relative sensitivity of the 2 test durations under similar conditions (i.e., feeding rates). Observing similar sensitivity, we settled on 14 d as the definitive test duration. Furthermore, we conducted experiments to determine how much food could be added to a given volume of water while minimally impacting dissolved metal recovery; a ratio of food dry mass to water volume (<0.005) achieved this. In the 14-d tests, we obtained a median lethal concentration and most sensitive chronic endpoint of 147 and 23 µg/L dissolved Ni (acute to chronic ratio [ACR] = 6.4), respectively, and 81 (mean value) and 10 µg/L dissolved Zn (ACR = 8.1), respectively. The acute values are orders of magnitude lower than previously published values for mayflies, probably most importantly due to the presence of dietary exposure but also potentially with some influence of organism age and test temperature. Environ Toxicol Chem 2020;39:1196-1206. © 2020 SETAC.

Declines in an abundant aquatic insect, the burrowing mayfly, across major North American waterways.

Seasonal animal movement among disparate habitats is a fundamental mechanism by which energy, nutrients, and biomass are transported across ecotones. A dramatic example of such exchange is the annual emergence of mayfly swarms from freshwater benthic habitats, but their characterization at macroscales has remained impossible. We analyzed radar observations of mayfly emergence flights to quantify long-term changes in annual biomass transport along the Upper Mississippi River and Western Lake Erie Basin. A single emergence event can produce 87.9 billion mayflies, releasing 3,078.6 tons of biomass into the airspace over several hours, but in recent years, production across both waterways has declined by over 50%. As a primary prey source in aquatic and terrestrial ecosystems, these declines will impact higher trophic levels and environmental nutrient cycling.

Bioaccumulation and Toxicity of Cadmium, Copper, Nickel, and Zinc and Their Mixtures to Aquatic Insect Communities.

We describe 2 artificial stream experiments that exposed aquatic insect communities to zinc (Zn), copper (Cu), and cadmium (year 2014) and to Zn, Cu, and nickel (year 2015). The testing strategy was to concurrently expose insect communities to single metals and mixtures. Single-metal tests were repeated to evaluate the reproducibility of the methods and year-to-year variability. Metals were strongly accumulated in sediments, periphyton, and insect (caddisfly) tissues, with the highest concentrations occurring in periphyton. Sensitive mayflies declined in metal treatments, and effect concentrations could be predicted effectively from metal concentrations in either periphyton or water. Most responses were similar in the replicated tests, but median effect concentration values for the mayfly Rhithrogena sp. varied 20-fold between the tests, emphasizing the difficulty comparing sensitivities across studies and the value of repeated testing. Relative to the single-metal responses, the toxicity of the mixtures was either approximately additive or less than additive when calculated as the product of individual responses (response addition). However, even less-than-additive relative responses were sometimes greater than responses to similar concentrations tested singly. The ternary mixtures resulted in mayfly declines at concentrations that caused no declines in the concurrent single-metal tests. When updating species-sensitivity distributions (SSDs) with these results, the mayfly responses were among the most sensitive 10th percentile of available data for all 4 metals, refuting older literature placing mayflies in the insensitive portion of metal SSDs. Testing translocated aquatic insect communities in 30-d artificial streams is an efficient approach to generate multiple species effect values under quasi-natural conditions that are relevant to natural streams. Environ Toxicol Chem 2020;39:812-833. Published 2020 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US government work, and as such, is in the public domain in the United States of America.

Assessing the toxicity of cell-bound microcystins on freshwater pelagic and benthic invertebrates.

Cyanobacterial harmful algal blooms dominated by Microcystis frequently produce microcystins, a family of toxins capable of inflicting harm to pelagic and benthic freshwater invertebrates. Research on the effect of microcystins on invertebrates is inconclusive; from one perspective, studies suggest invertebrates can coexist in toxic blooms; however, studies have also measured negative food-associated effects from microcystins. To test the latter perspective, we examined the reproduction, growth, and survival of laboratory-cultured Ceriodaphnia dubia, Daphnia magna, and Hexagenia spp. exposed to cell-bound microcystins through a series of life-cycle bioassays. Test organisms were exposed to a concentration gradient ranging from 0.5 µg L-1 to 300 µg L-1 microcystins, which corresponds to values typically found in freshwaters during bloom season. Lethal concentrations in C. dubia (LC50 = 5.53 µg L-1) and D. magna (LC50 = 85.72 µg L-1) exposed to microcystins were among the lowest recorded to date, and reproductive effects were observed at concentrations as low as 2.5 µg L-1. Length of D. magna was significantly impacted in microcystin treatments great than 2.5 µg L-1. No lethality or growth impairments were observed in Hexagenia. This information will improve our understanding of the risks posed by microcystins to food webs in freshwaters.

Effects of oil sands process water mixtures on the mayfly Hexagenia and field-collected aquatic macroinvertebrate communities.

Extraction of Canada's oil sands has created 1 billion m3 of tailings, which are stored in on-site tailings ponds. Due to limited storage capacity, the planned release of tailings into the surrounding environment may be required. This represents an environmental management challenge, as the tailings contain contaminants that are known toxins to aquatic communities. Of particular concern are naphthenic acids and their metallic counterparts, as they are the principal toxic components of tailings, are relatively soluble, and are persistent in aquatic environments. This study examines the acute toxicity of environmentally relevant 10:1 mixtures of two process water components: naphthenic acid and sodium naphthenate. We assess the effects of these simplified oil sands process water (OSPW) mixtures under planned and unplanned tailings release scenarios, using traditional and cutting-edge bioindicators for aquatic invertebrate taxa. We found that safe concentrations for mayflies and other aquatic macroinvertebrates were less than 1 mg/l, as no mayfly taxa survived repeated exposure to this dose in either the 48-h or 72-h acute toxicity test. In the 72-h test, no mayflies survived treatment levels greater than 0.5 mg sodium naphthenate/l. In the mesocosm study, even a 90% dilution of the OSPW mixture was not sufficient to protect sensitive macroinvertebrate communities. The results of this study highlight the potential environmental damage that will occur if OSPW is not carefully managed. This information will aid with the development of a management plan for oil sands tailings ponds, which will provide insight into the potential for process water release into the surrounding environment while conserving unique ecosystems downstream of development in the oil sands region.

Are sulfate effects in the mayfly Neocloeon triangulifer driven by the cost of ion regulation?

Elevated major ion concentrations in streams are commonly observed as a consequence of resource extraction, de-icing and other anthropogenic activities. Ecologists report biodiversity losses associated with increasing salinity, with mayflies typically being highly responsive to increases of different major ions. In this study, we evaluated the performance of the mayfly Neocloeon triangulifer reared for its entire larval phase in a gradient of sulfate concentrations. Two natural waters were amended with SO4 as a blend of CaSO4 and MgSO4 and exposures ranged from 5 to 1500 mg l-1 SO4. Survival (per cent successful emergence to the subimago stage) was significantly reduced at the highest SO4 concentration in both waters, while development was significantly delayed at 667 mg l-1 SO4 Final sub-adult body weights were consistent across treatments, except at the highest treatment concentration. Despite evidence for sulfate uptake rates increasing with exposure concentrations and not being saturated at even extremely high SO4 concentrations, total body sulfur changed little in subimagos. Together, these results suggest that elevated SO4 imposes an energetic demand associated with maintaining homeostasis that is manifested primarily as reduced growth rates and associated developmental delays. We identified two genes related to sulfate transport in N. trianguliferThis article is part of the theme issue 'Salt in freshwaters: causes, ecological consequences and future prospects'.

Dietary nutrient allocation to somatic tissue synthesis in emerging subimago freshwater mayfly Ephemera danica.

BACKGROUND: The relative importance of nutrients derived from different sources for tissue synthesis is crucial for predicting a species responds to changes in food availability. The ecological and physiological strategies that govern the incorporation and routing of nutrients for reproduction are often well understood. However, the role and adaptive value of both species and individual variation during early life-stage remain elusive. In freshwater systems, dietary nutrient allocation to somatic tissue should be favoured when dietary source peaks and resource limitation may hinder flexible resource allocation. We used carbon and nitrogen stable isotopes (δ13C and δ15N) to examine metabolic nutrient routing and resource allocation from four dietary sources used to biosynthesize three somatic tissues of emerging subimago Ephemera danica. Aquatic emerging insects, such as the mayfly E. danica, are well suited for such studies. This is because, while burrowing nymph phase is a detritivores feeders with several early life-stages of metamorphosis, adult insects do not feed during this period but do utilize energy. RESULTS: Constructed models to predict percent proportional contribution of source to tissue showed that terrestrial detritus was the dominant nutrient source for abdomen, head and wing with mean values of 57%, 65% and 73%, respectively. There was evidence for differential resource allocation, as insect partitioned periphyton and sediment (but also seston) elements for tissue synthesis. Utilizing individual-specimen based relationship in isotope value; we derived tissue specific isotopic niche estimates, for the different tissue-source combinations. CONCLUSIONS: Results indicate that tissue selection is crucial for isotopic ecological measurements in arthropods. Mayfly has long been used as bio-indicator of freshwater ecosystems and their larvae show rapid response to environmental changes. In light of the recent evidence of drastic reduction in flying insect mass in Germany, developing a system using isotopic tools to trace nutrient flow in this important taxon will assist conservation and management efforts.

Effects of macroconsumers on benthic communities: Rapid increases in dry-season accrual of calcium in a tropical karst stream.

Species loss from upper trophic levels can result in some major changes in community structure and ecosystem functions. Here, we experimentally excluded macroconsumers (e.g., fish and shrimp) in a Brazilian karst tropical stream during the dry season to investigate if their loss affected the accrual of calcium, dry mass (DM) and ash-free dry mass (AFDM) of sediment, benthic invertebrates, and chlorophyll-a. We found that the exclusion of macroconsumers decreased accrual of calcium. The absence of fish and shrimp may have promoted increased grazing by mayflies and snails in the electrified treatment as expressed by the depressed calcium accrual and shift in periphyton community composition. However, the exclusion of macroconsumers had no effect on DM and AFDM, chlorophyll-a, or total abundance of invertebrates. Our findings shed new light on the impact of macroconsumer loss and consequences for calcium accrual in karstic streams.

Reservoir longitudinal gradient promotes ordered losses on diversity and density of Ephemeroptera community / O gradiente longitudinal de um reservatório promove perdas ordenadas na diversidade e densidade da comunidade de Ephemeroptera

Abstract Reservoir operations alter, eliminate or restrain the natural hydrologic cycles. Biotic community has become subject to these non-cyclic events, responding by reducing the species diversity. Ephemeroptera species present distinct responses to environmental deterioration such that poses this assemblage between the most useful groups in biomonitoring programs. We hypothesized an alteration in beta diversity at the longitudinal species gradient, which will be influenced mainly by species losses between zones. Changes in temporal beta diversity is also expected, but the main drivers of such alterations will be the species turnover between the sampling period. Ephemeroptera community was monitored in nine sampling points from Itaipu Reservoir, where were installed three sets of substrates composed by a float and 2 wooden substrates. We took biological samples in triplicates monthly, from June-01 to August-02. Our initial hypothesis was partially supported and with significant variations only for spatial approach, between the Reservoir zones. The generated ordering from Non-Metric Dimensional Scale - NMDS - corroborated with spatial analyzes, with the formation of two groups along the gradient zonation of the reservoir. The temporal ordination showed no clear pattern. As expected, the contribution to beta diversity was different for our two approaches, such that the loss of species was more important along the spatial gradient and despite of no significant result, the species replacement was more important among months. The spatial results lead us to infer that differences in limnological characteristics between zones are important for determining differences in Ephemeroptera composition and can reflect the dependency degree of the species in relation to the lentic and sometimes-lotic conditions, mainly in the riverine zone of reservoirs. On the other hand, the absence of a temporal pattern can be result of chaotic variations in the physical and chemical attributes imposed by the reservoir operation, disrupting continuity of the biota and natural succession processes.

Resumo As operações de reservatórios alteram, eliminam ou restringem os ciclos hidrológicos naturais. A comunidade biótica torna-se sujeita a estes eventos não-cíclicos, respondendo com a redução da diversidade de espécies. As espécies de Ephemeroptera apresentam respostas distintas à deterioração ambiental, sendo o grupo muitousado em programas de biomonitoramento. Nós hipotetizamos uma alteração na diversidade beta das espécies no gradiente longitudinal, influenciada principalmente pelo componente aninhamento com a consequente perda de espécies entre as zonas do reservatório. Também são esperadas mudanças na diversidade beta entre os meses de amostragem, mas o principal fator dessas alterações será a substituição das espécies entre os períodos, em função dos rápidos ciclos de vida que o grupo apresenta. A comunidade Ephemeroptera foi monitorada em nove pontos amostrais do reservatório de Itaipu, onde foram instalados três conjuntos de substratos compostos por um flutuador e 2 substratos de madeira. Tomamos amostras biológicas em triplicatas mensalmente, de junho de 2001 a agosto de 2002. Nossa hipótese inicial foi parcialmente suportada, com variações significativas apenas na escala espacial. A ordenação gerada pelo Escalonamento Dimensional Não-Métrico - NMDS - corroborou com as análises espaciais, com a formação de dois grupos ao longo do gradiente de zonação do reservatório. A ordenação temporal não mostrou um padrão claro. Como esperado, a contribuição para a diversidade beta foi diferente para nossas duas abordagens, de modo que a perda de espécies foi mais importante ao longo do gradiente espacial e apesar de não haver resultado significativo, a substituição das espécies foi mais importante entre os meses de amostragem. Os resultados espaciais levam-nos a inferir que as diferenças nas características limnológicas entre as zonas são importantes para determinar as diferenças na composição de Ephemeroptera e podem refletir o grau de dependência das espécies em relação às condições lênticas e às vezes lóticas, principalmente na zona fluvial dos reservatórios. Por outro lado, a ausência de um padrão temporal pode ser resultado de variações caóticas nos atributos físicos e químicos impostos pela operação do reservatório, que interrompem a continuidade da biota e os processos de sucessão naturais.

Predicting mayfly recovery in acid mine-impaired streams using logistic regression models of in-stream habitat and water chemistry.

Mayflies (Order Ephemeroptera) require high quality water and habitat in streams to thrive, so their appearance after restoration is an indicator of ecological recovery. To better understand the importance of restoring in-stream habitat versus water chemistry for macroinvertebrate communities, we developed taxon-specific models of occurrence for five mayfly genera (Caenis, Isonychia, Stenonema, Stenacron, and Baetis) inhabiting streams in the Appalachian Mountains, USA. Presence/absence records from past decades were used to develop single and multiple logistic predictive models based on catchment characteristics (drainage area, gradient), in-stream habitat variables (e.g., substrate, channel morphology, pool and riffle quality), and water chemistry. Model performance was evaluated using (a) classification rates and Hosmer-Lemeshow values for test sets of data withheld from the original model-building dataset and (b) a field comparison of predicted versus observed mayfly occurrences at 53 sites in acid mine drainage-impaired watersheds in 2012. The classification accuracies of final models for Caenis, Stenacron, and Baetis ranged from 50 to 75%. In-stream habitat features were not significant predictor variables for these three taxa, only water chemistry. Models for Isonychia and Stenonema had higher classification rates (81%) and included both habitat and chemical variables. However, actual occurrences of Isonychia and Stenonema at study sites in 2012 were low, consistent with the calculated probability of occurrence (Po) < 0.60. Caenis occurred at test sites 35% of the time when the model predicted a Po > 0.40. Stenacron showed the greatest consistency of actual versus predicted occurrences, occurring at 56% of sites when the Po (based on pH and conductivity) was > 0.50 and only at 1 site when Po < 0.5. The results demonstrate how predictive models of individual indicator taxa could be valuable for evaluating the relative impacts of restoring physical habitat versus water chemistry during stream remediation.

Reservoir longitudinal gradient promotes ordered losses on diversity and density of Ephemeroptera community.

Reservoir operations alter, eliminate or restrain the natural hydrologic cycles. Biotic community has become subject to these non-cyclic events, responding by reducing the species diversity. Ephemeroptera species present distinct responses to environmental deterioration such that poses this assemblage between the most useful groups in biomonitoring programs. We hypothesized an alteration in beta diversity at the longitudinal species gradient, which will be influenced mainly by species losses between zones. Changes in temporal beta diversity is also expected, but the main drivers of such alterations will be the species turnover between the sampling period. Ephemeroptera community was monitored in nine sampling points from Itaipu Reservoir, where were installed three sets of substrates composed by a float and 2 wooden substrates. We took biological samples in triplicates monthly, from June-01 to August-02. Our initial hypothesis was partially supported and with significant variations only for spatial approach, between the Reservoir zones. The generated ordering from Non-Metric Dimensional Scale - NMDS - corroborated with spatial analyzes, with the formation of two groups along the gradient zonation of the reservoir. The temporal ordination showed no clear pattern. As expected, the contribution to beta diversity was different for our two approaches, such that the loss of species was more important along the spatial gradient and despite of no significant result, the species replacement was more important among months. The spatial results lead us to infer that differences in limnological characteristics between zones are important for determining differences in Ephemeroptera composition and can reflect the dependency degree of the species in relation to the lentic and sometimes-lotic conditions, mainly in the riverine zone of reservoirs. On the other hand, the absence of a temporal pattern can be result of chaotic variations in the physical and chemical attributes imposed by the reservoir operation, disrupting continuity of the biota and natural succession processes.

Jumping and the aerial behavior of aquatic mayfly larvae (Myobaetis ellenae, Baetidae).

Mayfly larvae generally are aquatic, but some madicolous taxa (i.e., living in thin water films) crawl over rocks within streams and waterfalls. When startled, these larvae can break the water film, jump, and enter an aerial phase of locomotion. Because mayfly larvae have been suggested as potential exemplars for the origin of insect wings as tracheal gills, and furthermore represent the most basal extant lineage of pterygotes, we analyzed jumping behavior and free-fall trajectories for one such species of mayfly (Myobaetis ellenae, Baetidae) in Costa Rica. Jumping was commonplace in this taxon, but was undirected and was characterized by body spinning at high angular velocities. No aerodynamic role for the tracheal gills was evident. By contrast, jumping by a sympatric species of bristletail (Meinertellus sp., Archaeognatha) consistently resulted in head-first and stable body postures during aerial translation. Although capable of intermittently jumping into the air, the mayfly larvae could neither control nor target their aerial behavior. By contrast, a stable body posture during jumps in adult bristletails, together with the demonstrated capacity for directed aerial descent in arboreal representatives of this order, support ancestrally terrestrial origins for insect flight within the behavioral context of either jumping or falling from heights.

Metabolomics reveal physiological changes in mayfly larvae (Neocloeon triangulifer) at ecological upper thermal limits.

Aquatic insects play critical roles in freshwater ecosystems and temperature is a fundamental driver of species performance and distributions. However, the physiological mechanisms that determine the thermal performance of species remain unclear. Here we used a metabolomics approach to gain insights into physiological changes associated with a short-term, sublethal thermal challenge in the mayfly Neocloeon triangulifer (Ephemeroptera: Baetidae). Larvae were subjected to a thermal ramp (from 22 to 30°C at a rate of 1°C/h) and metabolomics analysis (both Nuclear Magnetic Resonance (NMR) Spectroscopy and Gas Chromatography coupled Time-of-Flight Mass Spectrometry (GC-TOF-MS)) indicated that processes related to energetics (sugar metabolism) and membrane stabilization primarily differentiated heat treated larvae from controls. Limited evidence of anaerobic metabolism was observed in the heat treated larvae at 30°C, a temperature that is chronically lethal to larvae.

Physiological responses to short-term thermal stress in mayfly (Neocloeon triangulifer) larvae in relation to upper thermal limits.

Understanding species' thermal limits and their physiological determinants is critical in light of climate change and other human activities that warm freshwater ecosystems. Here, we ask whether oxygen limitation determines the chronic upper thermal limits in larvae of the mayfly Neocloeon triangulifer, an emerging model for ecological and physiological studies. Our experiments are based on a robust understanding of the upper acute (∼40°C) and chronic thermal limits of this species (>28°C, ≤30°C) derived from full life cycle rearing experiments across temperatures. We tested two related predictions derived from the hypothesis that oxygen limitation sets the chronic upper thermal limits: (1) aerobic scope declines in mayfly larvae as they approach and exceed temperatures that are chronically lethal to larvae; and (2) genes indicative of hypoxia challenge are also responsive in larvae exposed to ecologically relevant thermal limits. Neither prediction held true. We estimated aerobic scope by subtracting measurements of standard oxygen consumption rates from measurements of maximum oxygen consumption rates, the latter of which was obtained by treating with the metabolic uncoupling agent carbonyl cyanide-4-(trifluoromethoxy) pheylhydrazone (FCCP). Aerobic scope was similar in larvae held below and above chronic thermal limits. Genes indicative of oxygen limitation (LDH, EGL-9) were only upregulated under hypoxia or during exposure to temperatures beyond the chronic (and more ecologically relevant) thermal limits of this species (LDH). Our results suggest that the chronic thermal limits of this species are likely not driven by oxygen limitation, but rather are determined by other factors, e.g. bioenergetics costs. We caution against the use of short-term thermal ramping approaches to estimate critical thermal limits (CTmax) in aquatic insects because those temperatures are typically higher than those that occur in nature.

De novo transcriptome of the mayfly Cloeon viridulum and transcriptional signatures of Prometabola.

Mayflies (Ephemeroptera) display many primitive characters and a unique type of metamorphosis (Prometabola). However, information on the genomes and transcriptomes of this insect group is limited. The RNA sequencing study presented here generated the first de novo transcriptome assembly of Cloeon viridulum (Ephemeroptera: Baetidae), and compared gene expression signatures among the young larva (YL), mature larva (ML), subimago (SI), and imago (IM) stages of this mayfly. The transcriptome, based on 88 Gb of sequence data, comprised a set of 81,185 high quality transcripts. The number of differentially expressed genes (DEGs) in YL vs. ML, ML vs. SI, and SI vs. IM, was 4,825, 1,584, and 1,278, respectively, according to the reads per kilobase of transcript per million mapped reads analysis, assuming a false discovery rate <0.05 and a fold change >2. Gene enrichment analysis revealed that these DEGs were enriched in the "chitin metabolic process", "germ cell development", "steroid hormone biosynthesis", and "cutin, suberine, and wax biosynthesis" pathways. Finally, the expression pattern of a selected group of candidate signature genes for Prometabola, including vestigial, methoprene-tolerant, wingless, and broad-complex were confirmed by quantitative real time-PCR analysis. The Q-PCR analysis of larval, subimaginal, and imaginal stages of C. viridulum suggests that the development of mayflies more closely resembles hemimetamorphosis than holometamorphosis.

The nymph, habitat, and status of Eatonigenia in China (Ephemeroptera: Ephemeridae).

The nymphs of a new species of Eatonigenia have more modified labrum and smaller foreclaws than congeners, and the males have a more reddish-brown colored body, cerci, genitalia and more sclerotized projections of gonopores. These characters indicate a new species which is named E. zhangi sp. nov. The nymphs live in fine sandy/muddy substrates of large rivers.