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The sea cucumber (Apostichopus japonicus) has become a good model organism for studying environmentally induced aestivation in marine invertebrates. We hypothesized that mechanisms that arrest energy-expensive cell cycle activity would contribute significantly to establishing the hypometabolic state during aestivation. Cyclin A is a core and particularly interesting cell cycle regulator that functions in both the S phase and in mitosis. In the present study, negative relationships between miR-200-3p and AjCA expressions were detected at both the transcriptional and the translational levels during aestivation in A. japonicus. Dual-luciferase reporter assays confirmed the targeted location of the miR-200-3p binding site within the AjCA gene transcript. Furthermore, gain- and loss-of-function experiments were conducted in vivo with sea cucumbers to verify the interaction between miR-200-3p and AjCA in intestine tissue by qRT-PCR and Western blotting. The results show that the overexpression of miR-200-3p mimics suppressed AjCA transcript levels and translated protein production, whereas transfection with a miR-200-3p inhibitor enhanced both AjCA mRNA and AjCA protein in A. japonicus intestine. Our findings suggested a potential mechanism that reversibly arrests cell cycle progression during aestivation, which may center on miR-200-3p inhibitory control over the translation of cyclin A mRNA transcripts.

African lungfishes are ammonotelic in water. They can aestivate for long periods on land during drought. During aestivation, the gills are covered with dried mucus and ammonia excretion ceases. In fishes, ammonia excretion through the gills involves Rhesus glycoproteins (RhGP/Rhgp). This study aimed to obtain the complete cDNA coding sequences of rhgp from the gills of Protopterus annectens, and to determine their branchial mRNA and protein expression levels during the induction, maintenance and arousal phases of aestivation. Three isoforms of rhgp (rhag, rhbg and rhcg) were obtained in the gills of P. annectens. Their complete cDNA coding sequences ranged between 1311 and 1398 bp, coding for 436 to 465 amino acids with estimated molecular masses between 46.8 and 50.9 kDa. Dendrogramic analyses indicated that Rhag was grouped closer to fishes, while Rhbg and Rhcg were grouped closer to tetrapods. During the induction phase, the protein abundance of Rhag, but not its transcript level, was down-regulated in the gills, suggesting that there could be a decrease in the release of ammonia from the erythrocytes to the plasma. Furthermore, the branchial transcript levels of rhbg and rhcg decreased significantly, in preparation for the subsequent shutdown of gill functions. During the maintenance phase, the branchial expression levels of rhag/Rhag, rhbg/Rhbg and rhcg/Rhcg decreased significantly, indicating that their transcription and translation were down-regulated. This could be part of an overall mechanism to shut down branchial functions and save metabolic energy used for transcription and translation. It could also be regarded as an adaptive response to stop ammonia excretion. During the arousal phase, it is essential for the lungfish to regain the ability to excrete ammonia. Indeed, the protein abundance of Rhag, Rhbg and Rhcg recovered to the corresponding control levels after 1 day or 3 days of recovery from 6 months of aestivation. [ABSTRACT FROM AUTHOR]

Temperature-dependent development rate, percent diapause induction (hibernation at low temperature and aestivation at high temperature), and survival of diapausing larvae of Chilo partellus (Swinhoe, 1885) were examined on 13 constant temperatures ranging from 8 to 40 °C. Development of hibernating and aestivating larvae occurred from 10 to 25 °C and 27–38 °C, respectively. However, no development occurred at 8 °C and 40 °C. To determine actual thermal conditions that affect development and trigger both kind of diapause (hibernation and aestivation), various thermal parameters were estimated by fitting the development rate data to two linear (Ordinary equation and Ikemoto & Takai) models and thirteen non-linear models. The lower thermal thresholds ( T min ) for development of diapausing larvae of C. partellus were calculated as 9.60 °C and 10.29 °C using the ordinary linear model and Ikemoto & Takai model, respectively. Similarly, the thermal constants ( K ) estimated using the ordinary linear model was 333.33 degree-days and that estimated with Ikemoto & Takai model was 338.92 degree-days. Among the non-linear models, Lactin-2 followed by Lactin-1 were found to be the best as these models estimated the critical temperatures ( T min , T max and T opt ) similar to those of observed values. Conclusively, the Ikemoto & Takai linear model and Lactin-2 followed by Lactin-1 non-linear models are useful and efficient for describing temperature-dependent development and estimating the temperature thresholds of diapausing larvae of C. partellus . Our findings provided fundamental information for estimation of thermal requirement and temperature based development models for diapausing larvae of C. partellus . This information will be highly useful for predicting the occurrence, seasonal emergence, number of generations and population dynamics of C. partellus . [ABSTRACT FROM AUTHOR]

In arid environments, the source of the malaria mosquito populations that re-establish soon after first rains remains a puzzle and alternative explanations have been proposed. Using genetic data, we evaluated whether the early rainy season ( RS) population of Anopheles coluzzii is descended from the preceding late RS generation at the same locality, consistent with dry season ( DS) dormancy (aestivation), or from migrants from distant locations. Distinct predictions derived from these two hypotheses were assessed, based on variation in 738 SNPs in eleven A. coluzzii samples, including seven samples spanning 2 years in a Sahelian village. As predicted by the 'local origin under aestivation hypothesis,' temporal samples from the late RS and those collected after the first rain of the following RS were clustered together, while larger genetic distances were found among samples spanning the RS. Likewise, multilocus genotype composition of samples from the end of the RS was similar across samples until the following RS, unlike samples that spanned the RS. Consistent with reproductive arrest during the DS, no genetic drift was detected between samples taken over that period, despite encompassing extreme population minima, whereas it was detected between samples spanning the RS. Accordingly, the variance in allele frequency increased with time over the RS, but not over the DS. However, not all the results agreed with aestivation. Large genetic distances separated samples taken a year apart, and during the first year, within-sample genetic diversity declined and increased back during the late RS, suggesting a bottleneck followed by migration. The decline of genetic diversity followed by a mass distribution of insecticide-treated nets was accompanied by a reduced mosquito density and a rise in the mutation conferring resistance to pyrethroids, indicating a bottleneck due to insecticidal selection. Overall, our results support aestivation in A. coluzzii during the DS that is accompanied by long-distance migration in the late RS. [ABSTRACT FROM AUTHOR]

Elevated androgens and glucocorticoids displayed by males during the reproductive season have been proposed to mediate a possible trade-off between reproduction and immunocompetence. Anurans living in arid and semiarid environments display a strong seasonal reproduction, which could accentuate the variation in physiological, immunological, and behavioral parameters. We studied covariation between steroid plasma levels, morphometric variables associated with body condition and immunity, leukocyte profile, parasite load, and response to an immunological challenge across different phases of the annual life-history cycle of three anuran species from a Brazilian semiarid area. Our results showed a seasonal pattern of covariation among leukocyte parameters, kidney mass, and steroid plasma levels, with higher values measured during the reproductive season, particularly when males were sampled during calling activity. Moreover, these anurans showed a stronger response to an immunological challenge during the reproductive period. The immunosuppression during the dry period was particularly evident for the species that aestivate, indicating that the availability of energetic resources might be an important factor determining seasonal variation in inflammatory response. Intensity of the helminth infection was associated with eosinophil count but showed a more complex pattern with regard to androgens levels. These data emphasize that variations in the intensity of helminth infection might be more closely related to specific aspects of the immune response than to the general seasonal patterns of variation in steroid plasma levels, total circulating leukocytes, and inflammatory response. [ABSTRACT FROM AUTHOR]

The relationship of altitude, cardinal direction and plant species occupied by the sunn pest Eurygaster integriceps Puton (Heteroptera: Scutelleridae), during the aestivation period were determined. Studies were carried out on plant species ( Astragalus dipthterites Fenzl. , Noea spinosissim Moq. , Acantholimon sp. and Astragalus sp.), at 3 altitudes (1600 -1800; 1800-2000; 2000-2200 m), and in three directions (North, South, and East) from the overwintering sites on Nemrut Mountain, Adıyaman Turkey. According to the results, there were significant interactions of plant species x altitude, plant species x direction and altitude x direction. With respect to the species x altitude interaction, the most adults were found on N. spinosissim at altitudes of 1800-2000 m, and the least numbers were found on Astragalus sp. at the altitudes of 1600-1800 m. Considering the species x direction interaction, the most adults were found on N. spinosissim plants at the southern areas of overwintering sites, and the least numbers of adults were found on Astragalus sp. at the eastern sites. In the case of the altitude x direction interaction, the most adults were found in the southern areas of overwintering sites at 1800-2000 m, and the least numbers were found at the altitudes of 1600-1800 meters at the northern direction of overwintering sites. [ABSTRACT FROM AUTHOR]

Some animals can undergo a remarkable transition from active normal life to a dormant state called aestivation; entry into this hypometabolic state ensures that life continues even during long periods of environmental hardship. In this study, we aimed to identify those central nervous system (CNS) peptides that may regulate metabolic suppression leading to aestivation in land snails. Mass spectral-based neuropeptidome analysis of the CNS comparing active and aestivating states, revealed 19 differentially produced peptides; 2 were upregulated in active animals and 17 were upregulated in aestivated animals. Of those, the buccalin neuropeptide was further investigated since there is existing evidence in molluscs that buccalin modulates physiology by muscle contraction. The Theba pisana CNS contains two buccalin transcripts that encode precursor proteins that are capable of releasing numerous buccalin peptides. Of these, Tpi-buccalin-2 is most highly expressed within our CNS transcriptome derived from multiple metabolic states. No significant difference was observed at the level of gene expression levels for Tpi-buccalin-2 between active and aestivated animals, suggesting that regulation may reside at the level of post-translational control of peptide abundance. Spatial gene and peptide expression analysis of aestivated snail CNS demonstrated that buccalin-2 has widespread distribution within regions that control several physiological roles. In conclusion, we provide the first detailed molecular analysis of the peptides and associated genes that are related to hypometabolism in a gastropod snail known to undergo extended periods of aestivation. [ABSTRACT FROM AUTHOR]

African lungfishes can aestivate and remain torpid without food and water for years, but disuse muscle atrophy is not prominent during aestivation. This study aimed to clone myostatin ( mstn/Mstn), a factor associated with disuse muscle atrophy in mammals, from the skeletal muscle of the African lungfish Protopterus annectens, and to determine its mRNA expression level and protein abundance therein during the induction, maintenance, and arousal phases of aestivation. The complete coding cDNA sequence of mstn comprised 1128 bp, encoding for 376 amino acids with an estimated molecular mass of 42.9 kDa. It was grouped together with Mstn/MSTN of coelacanth and tetrapods in a clade separated from teleost Mstn. After 6 days (the induction phase) of aestivation, the mstn transcript level in the muscle increased significantly, while the protein abundance of Mstn remained comparable to the control. Following that, a significant increase in the expression levels of mstn/Mstn occurred on day 12 (the early maintenance phase) of aestivation. After 6 months of aestivation (the prolonged maintenance phase), the expression levels of mstn/Mstn returned to control levels, indicating the possible impediment of a drastic increase in muscle degradation to prevent muscle atrophy. During 1-3 days of arousal from aestivation, the expression levels of mstn/Mstn in the muscle remained comparable to the control. Hence, tissue reconstruction/regeneration of certain organs might not involve the mobilization of amino acids from the muscle during the early arousal. These results provide insights into how aestivating P. annectens regulates the expression of mstn/Mstn possibly to ameliorate disuse muscle atrophy. [ABSTRACT FROM AUTHOR]

Hemlock woolly adelgid (HWA), Adelges tsugae Annand, an invasive insect native to the Pacific Northwest and Asia, is responsible for widespread health decline and mortality of native hemlocks (Tsuga spp.) in the eastern United States. Shading and fertilizer has been found to affect the survival and health of both HWA and hemlocks. These abiotic factors have been studied separately but not in combination. In this three year study, eastern hemlock trees (1-2 m tall) were treated with pruning, fertilizer, and shade to determine their effects on hemlock tree health and HWA survival and density. Shade cloths were erected over individual trees, granulated fertilizer was applied, and trees were pruned annually. The total number of HWA were counted during the sistens and progrediens adult stages on the low, mid, and high branches on the north, east, south, and west sides of each tree for three years. Survival of aestivating sistens was recorded in artificially, naturally, and unshaded hemlocks. The mean of percent tips alive, branches alive, and foliage density was used to calculate a hemlock health index (scale of 0-100). Shade cloth reduced solar radiation to the trees to levels similar to a naturally-forested hemlock canopy, but did not alter temperature. Trees exposed to shade alone and shade plus fertilizer maintained the greatest HWA density. On unshaded trees, branches on the west side of the tree had lower HWA densities and branches high on the tree had the lowest HWA densities. Pruning plus fertilizer and shading plus fertilizer reduced tree health. Shaded trees had reduced branchlet new growth length. Survival of summer aestivating sistens was nearly twice the survival under artificially- and naturally-shaded trees compared to unshaded trees. There was an inverse density-dependent survival response for aestivating HWA under artificially-shaded and unshaded trees but not naturally-shaded trees. Unshaded hemlock trees had lower HWA densities due to increased mortality of summer aestivating sistens. Unshaded trees had better health and longer new growth branchlets due to increased exposure to solar radiation and lower HWA densities. Silvicultural thinning of hemlocks in forest stands could increase direct sunlight reaching the trees and help decrease HWA densities and improve hemlock health. [ABSTRACT FROM AUTHOR]

Background: Variation in longevity has long been of interest in vector biology because of its implication in disease transmission through vectorial capacity. Recent studies suggest that Anopheles coluzzii adults persist during the ~7 month dry season via aestivation. Recently there has been a growing body of evidence linking dietary restriction and low ratio of dietary protein to carbohydrate with extended longevity of animals. Here, we evaluated the effects of dietary restriction and the protein : carbohydrate ratio on longevity of An. coluzzii. Results: In our experiment, we combined dietary regimes with temperature and relative humidity to assess their effects on An. coluzzii longevity, in an attempt to simulate aestivation under laboratory conditions. Our results showed significant effects of both the physical and the dietary variables on longevity, but that diet regimen had a considerably greater effect than those of the physical conditions. Higher temperature and lower humidity reduced longevity. At 22 °C dietary protein (blood) shortened longevity when sugar was not restricted (RH = 85%), but extended longevity when sugar was restricted (RH = 50%). Conclusions: Dietary restriction extended longevity in accord with predictions, but protein : carbohydrate ratio had a negligible effect. We identified conditions that significantly extend longevity in malaria vectors, however, the extent of increase in longevity was insufficient to simulate aestivation. [ABSTRACT FROM AUTHOR]

It is difficult to obtain non-destructive information on the seasonal dynamics of earthworms in northern forest soils. To overcome this, we used a Rhizotron facility to compile 7 years of data on the activity of anecic ( Lumbricus terrestris) and endogeic ( Aporrectodea caliginosa complex) earthworms in two contrasting soil/plant community types. We hypothesized that L. terrestris burrows would be used for longer than a typical L. terrestris lifetime, and that the distribution and activity pattern of the two earthworm species would respond differently to changes in soil moisture and temperature. For 7 years we recorded earthworm distribution and activity state bi-weekly to a depth of 1.5 m, tracked L. terrestris burrows using images captured annually, and measured soil temperature and moisture. Activity and vertical distribution of earthworms was closely linked to earthworm species and soil temperature in the fall, winter and spring. Lumbricus terrestris typically remained active through the winter, whereas the A. caliginosa complex was more likely to enter an aestivation period. Activity of all earthworms decreased substantially in July and August when soil temperature was at its highest and soil moisture at its lowest for the year. Most L. terrestris burrows were used continuously and moved very little during the 7-year study, likely creating spatiotemporally stable hotspots of soil resources. The different patterns of response of these species to soil temperature and moisture suggests that endogeic earthworms are more likely than anecic earthworms to adjust activity states in response to climate change mediated shifts in soil moisture and temperature. [ABSTRACT FROM AUTHOR]

The sea cucumber, Apostichopus japonicus , is a good model for studying environmentally-induced aestivation by a marine invertebrate. One of the central requirements of aestivation is the repression of energy-expensive cellular processes such as cell cycle progression. The present study identified the gene structure of the cell cycle regulator, cyclin B , and detected the expression levels of this gene over three stages of the annual aestivation-arousal cycle. Furthermore, the DNA methylation characteristics of cyclin B were analyzed in non-aestivation and deep-aestivation stages of sea cucumbers. We found that the cyclin B promoter contains a CpG island, three CCAAT-boxes and three cell cycle gene homology regions (CHRs). Application of qRT-PCR analysis showed significant downregulation of cyclin B transcript levels during deep-aestivation in comparison with non-aestivation in both intestine and longitudinal muscle, and these returned to basal levels after arousal from aestivation. Methylation analysis of the cyclin B core promoter revealed that its methylation level showed significant differences between non-aestivation and deep-aestivation stages ( p < 0.05) and interestingly, a positive correlation between Cyclin B transcripts expression and methylation levels of the core promoter was also observed. Our findings suggest that cell cycle progression may be reversibly arrested during aestivation as indicated by the changes in cyclin B expression levels and we propose that DNA methylation is one of the regulatory mechanisms involved in cyclin B transcriptional variation. [ABSTRACT FROM AUTHOR]

African lungfishes can undergo long periods of aestivation on land during drought. During aestivation, lungfishes are confronted with desiccation and dehydration, and their gills become non-functional and covered with a thick layer of dried mucus. Aquaporins (Aqps) are a superfamily of integral membrane proteins which generally facilitate the permeation of water through plasma membranes. This study aimed to obtain the complete cDNA coding sequences of aqp1 and aqp3 from the gills of Protopterus annectens, and to determine their branchial mRNA and protein expression levels during the induction, maintenance and arousal phases of aestivation. Dendrogramic analyses of the deduced Aqp1 and Aqp3 amino acid sequences of P. annectens revealed their close relationships with those of Latimeria chalumnae and tetrapods. During the induction phase, there were significant decreases in the transcript levels of aqp1 and aqp3 in the gills of P. annectens, but the branchial Aqp1 and Aqp3 protein abundance remained unchanged. As changes in transcription might precede changes in translation, this could be regarded as an adaptive response to decrease the protein abundance of Aqp1 and Aqp3 in the subsequent maintenance phase of aestivation. As expected, the branchial transcript levels and protein abundance of aqp1/Aqp1 and aqp3/Aqp3 were significantly down-regulated during the maintenance phase, probably attributable to the shutdown of branchial functions and the cessation of volume regulation of branchial epithelial cells. Additionally, these changes could reduce the loss of water through branchial epithelial surfaces, supplementing the anti-desiccating property of the dried mucus. Upon arousal, it was essential for the lungfish to restore branchial functions. Indeed, the protein abundance of Aqp1 recovered partially, with complete recovery of mRNA expression level and protein abundance of Aqp3, in the gills of P. annectens after 3 days of arousal. These results provide insights into how P. annectens regulates branchial Aqp expression to cope with desiccation and rehydration during different phases of aestivation. [ABSTRACT FROM AUTHOR]

Dispersal from the release site and high mortality are two main factors that contribute to low retention of hard-released translocated wildlife. A soft-release translocation is a method that may increase the likelihood of translocation success because individuals are forced to acclimatize and become familiar to the new release site prior to the translocation. The objective of this study was to compare the movement patterns and retention rate of resident and translocated Egyptian tortoises that were translocated just prior to the start of the aestivation season and therefore were forced to aestivate at the release site (a forced in-activity soft-release). The retention rate of translocated tortoises, the proportion of tortoises that remained at the release site and alive by the end of the study, was 71% (5/7) compared to a retention rate of 100% for resident tortoises. There was no significant difference in the minimum convex polygon area, total distance moved, or the number of relocations during the activity season between resident and translocated tortoises. Our results suggest that a forced in-activity soft release consisting of translocating Egyptian tortoises just before the aestivation season may be effective in minimizing dispersal from the release site. [ABSTRACT FROM AUTHOR]

Abstract Rapid evolution has rarely been assessed in biological control systems despite the similarity with biological invasions, which are widely used as model systems. We assessed post‐introduction climatic adaptation in a population of Longitarsus jacobaeae, a biological control agent of Jacobaea vulgaris, which originated from a low‐elevation site in Italy and was introduced in the USA to a high‐elevation site (Mt. Hood, Oregon) in the early 1980s. Life‐history characteristics of beetle populations from Mt. Hood, from two low‐elevation sites in Oregon (Italian origin) and from a high‐elevation site from Switzerland were compared in common gardens. The performance of low‐ and high‐elevation populations at a low‐ and a high‐elevation site was evaluated using reciprocal transplants. The results revealed significant changes in aestival diapause and shifts in phenology in the Mt. Hood population, compared with the low‐elevation populations. We found increased performance of the Mt. Hood population in its home environment compared with the low‐elevation populations that it originated from. The results indicate that the beetles at Mt. Hood have adapted to the cooler conditions by life‐history changes that conform to predictions based on theory and the phenology of the cold‐adapted Swiss beetles.

At the south western border of its extensive distribution, the multivoltine large white butterfly, Pieris brassicae L., is exceptional in undergoing summer diapause or aestivation. In all other regions investigated, P. brassicae pupae only hibernate. The transitional zone from non-aestivating to aestivating populations is a geographically stable region south of the Pyrenees. The restriction of this response to this region cannot be accounted for in terms of genetics as aestivation is intermediately inherited, with the heritability (h2) of aestivation in inbreeding lines between 0.35 and 0.77. Two hypotheses are presented to explain why this species does not aestivate in more northern regions. First, aestivation is a behaviour that serves to synchronize generations in areas where this species produces a high number of generations per year. Second, aestivation reduces the incidence of parasitism suffered by the butterfly by desynchronizing its life cycle from that of its main parasitoid, Cotesia glomerata. The two hypotheses are not mutually exclusive and both seem to be adaptive where the species is multivoltine.

Achatina fulica Bowdich, 1822 is an african snail that is invasive in different parts of the world, being characterized mainly by its high adaptability. Our objective was to evaluate the influence of abiotic factors, such as temperature and substrate, on the egg eclosion, adult and juvenile reaction to different abiotics factors, and the resistance of A. fulica to popular methods of control. Three studies were made, including 19 laboratory experiments and observations of free animals in the municipal district of Guaraqueçaba, southern Brazil. Eggs were characterized as the most fragile phase, whereas the juveniles and adults were resistant to the immersion in fresh and salt water and little resistant to temperature variation and salt, using burying, aestivation, and muscular force as defense strategies. Those strategies should be considered in management actions and in orientating popular methods of control.

Effect of pre-diapause temperature on summer and winter diapause intensity was examined under both laboratory and field conditions. Under short photoperiods of 8L : 16D and 10L : 14D, all pupae entered diapause at 15, 18 and 20°C and the incidence of diapause dropped to 82.3% and 85.5% at 22°C, respectively. Under long photoperiods of 14L : 10D and 16L : 8D, the incidence of diapause decreased with increasing temperature and there were significant differences among temperatures. The incidence of diapause at 16L : 8D was significantly lower than that under14L : 10D at 20 and 22°C. By transferring diapause pupae induced under various temperatures (18, 20 and 22°C) at a short day of 10L : 14D or a long day of 14L : 10D, to 12.5L : 11.5D, 20°C, the duration of summer diapause induced under 22°C (mean 76.1 days) was significantly shorter than those under 20°C (mean 85.9 days) and 18°C (mean 90.9 days), showing that the incidence of summer diapause was positively linked to the intensity of summer diapause; whereas the duration of winter diapause induced under 10L : 14D was similar at 18°C (89.2 days), 20°C (88.7 days) and 22°C (89.2 days) and there were no significant differences. Field experiments also showed that the high rearing temperatures significantly decreased the incidence and intensity of summer diapause, but had no significant affect on the intensity of winter diapause. When the naturally aestivating pupae from the first spring generation (formed on 24 April) and second spring generation (formed on 15 May) were kept under summer conditions, the diapause duration of the first generation lasted for 107-166 days (mean 146 days), about twenty days longer than that of the second generation [lasted for 92-151 days (mean 126 days)]. All results reveal that the sensitivity to temperature prior to aestivation and hibernation was quite different.

Effect of photoperiod on the duration of summer and winter diapause was investigated in the cabbage butterfly, Pieris melete. By keeping naturally induced aestivating and hibernating pupae under various photoperiods, it was shown that diapause duration of aestivating pupae was significantly longer at long than at short daylengths, whereas diapause duration of hibernating pupae was significantly shorter at long than at short daylengths, suggesting both aestivating and hibernating pupae require opposite photoperiodic signals to promote diapause development. By transferring diapausing pupae, induced under various photoperiods, to 20°C with a naturally changing summer daylength, the diapause induced by short daylengths was easier to terminate than diapause induced by long daylengths. When naturally induced aestivating and hibernating pupae were kept under natural conditions, aestivating pupae had a long diapause (mean 155 days) and wide range of emergence (90 days), whereas hibernating pupae had a short diapause (mean 105 days) and a relatively synchronized emergence (lasted 30 days). Finally, the ecological significance of photoperiodic regulation of diapause duration is discussed.

Bone mass and skeletal muscle mass are controlled by factors such as genetics, diet and nutrition, growth factors and mechanical stimuli. Whereas increased mechanical loading of the musculoskeletal system stimulates an increase in the mass and strength of skeletal muscle and bone, reduced mechanical loading and disuse rapidly promote a decrease in musculoskeletal mass, strength and ultimately performance (i.e. muscle atrophy and osteoporosis). In stark contrast to artificially immobilised laboratory mammals, animals that experience natural, prolonged bouts of disuse and reduced mechanical loading, such as hibernating mammals and aestivating frogs, consistently exhibit limited or no change in musculoskeletal performance. What factors modulate skeletal muscle and bone mass, and what physiological and molecular mechanisms protect against losses of muscle and bone during dormancy and following arousal? Understanding the events that occur in different organisms that undergo natural periods of prolonged disuse and suffer negligible musculoskeletal deterioration could not only reveal novel regulatory factors but also might lead to new therapeutic options. Here, we review recent work from a diverse array of species that has revealed novel information regarding physiological and molecular mechanisms that dormant animals may use to conserve musculoskeletal mass despite prolonged inactivity. By highlighting some of the differences and similarities in musculoskeletal biology between vertebrates that experience disparate modes of dormancy, it is hoped that this Review will stimulate new insights and ideas for future studies regarding the regulation of atrophy and osteoporosis in both natural and clinical models of muscle and bone disuse. [ABSTRACT FROM AUTHOR]

The state of metabolic dormancy has fascinated people for hundreds of years, leading to research exploring the identity of natural molecular components that may induce and maintain this state. Many animals lower their metabolism in response to high temperatures and/or arid conditions, a phenomenon called aestivation. The biological significance for this is clear; by strongly suppressing metabolic rate to low levels, animals minimize their exposure to stressful conditions. Understanding blood or hemolymph metabolite changes that occur between active and aestivated animals can provide valuable insights relating to those molecular components that regulate hypometabolism in animals, and how they afford adaptation to their different environmental conditions. In this study, we have investigated the hemolymph metabolite composition from the land snail Theba pisana , a remarkably resilient mollusc that displays an annual aestivation period. Using LC-MS-based metabolomics analysis, we have identified those hemolymph metabolites that show significant changes in relative abundance between active and aestivated states. We show that certain metabolites, including some phospholipids [e.g. LysoPC(14:0)], and amino acids such as l -arginine and l -tyrosine, are present at high levels within aestivated snails. Further investigation of our T. pisana RNA-sequencing data elucidated the entire repertoire of phospholipid-synthesis genes in the snail digestive gland, as a precursor towards future comparative investigation between the genetic components of aestivating and non-aestivating species. In summary, we have identified a large number of metabolites that are elevated in the hemolymph of aestivating snails, supporting their role in protecting against heat or desiccation. [ABSTRACT FROM AUTHOR]

Marine invertebrate aestivation is a unique strategy for summer survival in response to hot marine conditions. The sea cucumber, Apostichopus japonicus , is an excellent model marine invertebrate for studies of environmentally-induced aestivation. In the present study, we used a tandem mass tag (TMT)-coupled LC-MS/MS approach to identify and quantify the global proteome expression profile over the aestivation-arousal cycle of A. japonicus . A total of 3920 proteins were identified from the intestine of sea cucumber. Among them, 630 proteins showed significant differential expression when comparing three conditions of sea cucumbers: non-aestivating (active), deep-aestivation (at least 15 days of continuous aestivation), and arousal after aestivation (renewed moving and feeding). Sea cucumbers in deep aestivation showed substantial differentially expressed proteins (143 up-regulated and 267 down-regulated proteins compared with non-aestivating controls). These differentially expressed proteins suggested that protein and phospholipid probably are major fuel sources during hypometabolism and a general attenuation of carbohydrate metabolism was observed during deep aestivation. Differentially expressed proteins also provided the first global picture of a shift in protein synthesis, protein folding, DNA binding, apoptosis, cellular transport and signaling, and cytoskeletal proteins during deep aestivation in sea cucumbers. A comparison of arousal from aestivation with deep aestivation, revealed a general reversal of the changes that occurred in aestivation for most proteins. Western blot detection further validated the significant up-regulation of HSP70 and down-regulation of methyltransferase-like protein 7A-like in deep-aestivation. Our results suggest that there is substantial post-transcriptional regulation of proteins during the aestivation-arousal cycle in sea cucumbers. [ABSTRACT FROM AUTHOR]

Climate change is causing prolonged drying in many seasonal wetlands, including urban wetlands, potentially affecting aquatic invertebrates that take refuge in wetland sediment during dry periods and thereby threatening wetland biodiversity. We collected sediment from two habitats: open water (OW) and fringing trees (FT), in eight urban wetlands after seasonal inundation had ended. Both habitats are inundated during winter-spring and dry in summer-autumn. Each sediment sample was divided into subsamples. One set of subsamples were inundated in the laboratory to test the hypothesis that emerging invertebrate assemblages would differ between OW and FT sediments. Another set of subsamples was dried, stored for a year, and inundated to test the hypothesis that prolonged drying would reduce the abundance and taxa richness of emerging invertebrates. The composition of emerging invertebrate assemblages differed between habitats, with more amphibious species found in FT sediment. Invertebrate responses to prolonged drying and storage varied among species: for some, effects depended on habitat type, while others delayed emergence or showed no response. Microcrustacean abundance was unaffected by drying, suggesting that their productivity during refilling may resist drier water regimes. Surface temperatures of dry sediment are cooler beneath FT, and this sediment has higher organic matter, holds more water and is less dense than OW sediment; and FT sediment remained cooler than OW sediment in the laboratory, despite the absence of shading. Fringing trees may therefore provide a refuge for some freshwater invertebrates relying on dormant stages in the sediment to survive drying in urban wetlands. [ABSTRACT FROM AUTHOR]

Aestivation is a state of dormancy in diverse animal species characterized by a depressed metabolism. High temperatures in the summer trigger aestivation in the adult sea cucumber, Apostichopus japonicus. In the present study, the glycolytic suppression in aestivating A. japonicus was verified by metabolite quantification and activity and a gene expression analysis of rate-limiting enzymes (hexokinase and pyruvate kinase). Significant decreases in glucose concentrations were discovered in the respiration tree ( p < 0.01), intestine ( p < 0.05) and muscle ( p < 0.01) of aestivating A. japonicus; likewise, over this period, pyruvic acid concentrations significantly decreased in the respiration tree ( p < 0.05) and intestine ( p < 0.01), but were unchanged in muscle. Enzymatic activity variation of hexokinase ( Aj-HK) and pyruvate kinase ( Aj-PK) in aestivating A. japonicus revealed that Aj-HK remained at high levels in the respiration tree and muscle but significantly decreased in intestine ( p < 0.01); the activity of Aj-PK also declined significantly in intestine ( p < 0.01), whereas it increased significantly in the respiration tree ( p < 0.05) and muscle ( p < 0.01). The gene sequences of Aj-HK and Aj-PK were identified, and their transcriptional quantification revealed that the expression of Aj-HK was depressed in all of the examined tissues, whereas Aj-PK expression was depressed in muscle only. These results suggest that glycolytic suppression occurred in aestivating A. japonicus, especially in the intestine, and transcriptional and post-transcriptional modifications are involved in the regulation of glycolytic suppression in this marine species. [ABSTRACT FROM AUTHOR]

Schistosomiasis vector snails are subjected to extreme seasonal changes, particularly in ephemeral rivers and lentic waterbodies. In the tropics, aestivation is one of the adaptive strategies for survival and is used by snails in times of extremely high temperatures and desiccation. Aestivation therefore plays an important role in maintaining the transmission of schistosomiasis. This review assesses the possible impacts of climate change on the temporal and spatial distribution of schistosomiasis-transmitting snails with special emphasis on aestivation, and discusses the effect of schistosome infection on aestivation ability. The impacts of parasite development on snails, as well as physiological changes, are discussed with reference to schistosomiasis transmission. This review shows that schistosome-infected snails have lower survival rates during aestivation, and that those that survive manage to get rid of the infection. In general, snail aestivation ability is poor and survival chances diminish with time. Longer dry periods result in fewer, as well as uninfected, snails. However, the ability of the surviving snails to repopulate the habitats is high. [ABSTRACT FROM AUTHOR]

With water temperature increasing in summer, large-sized sea cucumbers ( Apostichopus japonicus ) generally enter aestivation, while there still occurs a small portion of individuals that do not enter aestivation and remain active and ingesting. In this study we investigated the differences in immuno- and metabolic enzyme activities and metabolites between non-aestivating and aestivating groups of A. japonicus . Animals (wet weight > 150 g) were collected from July to October in a seawater pond, north China. The individuals that voided feces, or had intestine contents after dissection, were defined as non-aestivating group, whereas those without feces or intestine contents were defined as aestivating group. Relative intestine mass (RIM), immuno-enzyme activities (SOD, CAT, T-AOC) and metabolic enzyme activities (PK, HK, MDH, LDH) of the body wall, and metabolites in intestine in both groups were determined. Results showed that temperature had significant effects on RIM and all enzyme activities in the non-aestivating group ( P < 0.05). However, the significant effects of temperature on the T-AOC, PK and LDH activities were not found in the aestivating group ( P > 0.05). Meanwhile, the RIM and all enzyme activities in the non-aestivating group were significantly higher than those in the aestivating group before September 10. The metabolites including glutamate, 3-aminoisobutyrate, aspartate, choline, phosphocholine, and taurine were abundant in the aestivating group, whereas 12 kinds of amino acids, glucose, glycogen, dimethylamine, inosine, and homarine were abundant in the non-aestivating group. It was obvious that metabolism of the non-aestivating group was more active than that of the aestivating group. This study provided new insights for understanding the differences in intestine degradation, enzyme activities and metabolites between non-aestivating and aestivating A. japonicus under high temperature in summer. Meanwhile, the results might be helpful for high temperature resistant sea cucumber selection in aquaculture. Statement of relevance We state that the work has not been published and is not being considered for published elsewhere. [ABSTRACT FROM AUTHOR]

Hypometabolism is a physiological state of dormancy entered by many animals in times of environmental stress. There are gaps in our understanding of the molecular components used by animals to achieve this metabolic state. The availability of genomic and transcriptome data can be useful to study the process of hypometabolism at the molecular level. In this study, we use the land snail Theba pisana to identify peptides that may be involved in the hypometabolic state known as aestivation. We found a total of 22 neuropeptides in the central nervous system (CNS) that were differentially produced during activity and aestivation based on mass spectral-based neuropeptidome analysis. Of these, 4 were upregulated in active animals and 18 were upregulated in aestivation. A neuropeptide known to regulate muscle contractions in a variety of molluscs, the small cardioactive peptide A (sCAP A ), and a peptide of yet unknown function (termed Aestivation Associated Peptide 12) were chosen for further investigation using temporal and spatial expression analysis of the precursor gene and peptide. Both peptides share expression within regions of the CNS cerebral ganglia and suboesophageal ganglia. Relative transcript abundance suggests that regulation of peptide synthesis and secretion is post-transcriptional. In summary, we provide new insights into the molecular basis of the regulation of aestivation in land snails through CNS peptide control. [ABSTRACT FROM AUTHOR]

The African malaria mosquitoes Anopheles gambiae and Anopheles coluzzii range over forests and arid areas, where they withstand dry spells and months-long dry seasons, suggesting variation in their desiccation tolerance. We subjected a laboratory colony (G3) and wild Sahelian mosquitoes during the rainy and dry seasons to desiccation assays. The thoracic spiracles and amount and composition of cuticular hydrocarbons (CHCs) of individual mosquitoes were measured to determine the effects of these traits on desiccation tolerance. The relative humidity of the assay, body water available, rate of water loss and water content at death accounted for 88% of the variation in desiccation tolerance. Spiracle size did not affect the rate of water loss or desiccation tolerance of the colony mosquitoes, as was the case for the total CHCs. However, six CHCs accounted for 71% of the variation in desiccation tolerance and three accounted for 72% of the variation in the rate of water loss. Wild A. coluzzii exhibited elevated desiccation tolerance during the dry season. During that time, relative thorax and spiracle sizes were smaller than during the rainy season. A smaller spiracle size appeared to increase A. coluzzii's desiccation tolerance, but was not statistically significant. Seasonal changes in CHC composition were detected in Sahelian A. coluzzii. Stepwise regression models suggested the effect of particular CHCs on desiccation tolerance. In conclusion, the combination of particular CHCs along with the total amount of CHCs is a primary mechanism conferring desiccation tolerance in A. coluzzii, while variation in spiracle size might be a secondary mechanism. [ABSTRACT FROM AUTHOR]

Climate change is modifying the timing of the onset of winter rainfall in southern Australia, at times creating brief inundation events in seasonal wetlands, termed 'false starts'. False starts may cause abortive hatchings of fauna emerging from sediment egg banks because wetlands dry out before invertebrates can complete their life cycle or reach a drying-resistant life stage., A laboratory emergence experiment was used to determine whether the abortive hatching caused by false starts alters assemblage composition in the subsequent hydroperiod and whether the length of the dry period following a false start alters subsequent assemblage composition. Sediment for the experiment was collected from Lake Joondalup South, Swan Coastal Plain (SCP), Western Australia, because it has a relatively diverse assemblage of desiccation-resistant invertebrate propagules. Most wetlands on the SCP are seasonal and groundwater fed and the region has a mediterranean-type climate, Two different habitat types, open water (OW) and fringing trees (FT), with distinct freshwater invertebrate assemblages are commonly found in SCP wetlands. We repeated the experiment in sediment from both habitats to determine whether false starts had the same effect on the two assemblages. Replicate sediment samples from both habitats were placed in microcosms randomly allocated to treatments or controls. To simulate false starts of differing dry-period duration, treatments were inundated for 5 days, then allowed to dry out for different time periods (10, 20 and 30 days) and then inundated for 5 days. Controls were inundated for time periods equivalent to the total duration of each false-start treatment (20, 30 and 40 days)., FT sediment had higher organic matter and moisture content than OW sediment. The composition of the emerging assemblage differed between habitats, and emergence was slower from OW than FT sediment. Abortive hatching followed the false start in OW sediment, but subsequently the same assemblage emerged, showing reliance on the egg bank to resupply lost populations. Abortive hatching was not observed in FT sediment, where invertebrates survived drying during the false start, continuing to develop for up to 30 days without surface water, because those sediments retained moisture., Provided that winter-spring hydroperiods continue to inundate OW for several months, these results indicate that invertebrates will be able to complete their life cycles and replenish egg banks following abortive hatching, demonstrating resilience to false starts. False starts to winter inundation rarely occur more than a month prior to the start of 'true' winter rains, so FT assemblages are resistant to false starts, showing the ability to survive dry periods of up to 30 days. Furthermore, survival in damp FT sediment also gave these populations a 'head start', because the surviving assemblage was identical to the continuously inundated control., Assemblages emerging from beneath FT may potentially recolonise inundated OW habitat. So far, changes to SCP seasonal-wetland hydroperiods will be within the coping range of their invertebrate assemblages. As FT assemblages were more resistant to false starts, restoration schemes that increase shading by fringing vegetation should be encouraged. [ABSTRACT FROM AUTHOR]

The terrestrial Roman snail Helix pomatia has successfully adapted to strongly fluctuating conditions in its natural soil habitat. Part of the snail’s stress defense strategy is its ability to express Metallothioneins (MTs). These are multifunctional, cysteine-rich proteins that bind and inactivate transition metal ions (Cd2+, Zn2+, Cu+) with high affinity. In Helix pomatia a Cadmium (Cd)-selective, inducible Metallothionein Isoform (CdMT) is mainly involved in detoxification of this harmful metal. In addition, the snail CdMT has been shown to also respond to certain physiological stressors. The aim of the present study was to investigate the physiological and diurnal variability of CdMT gene expression in snails exposed to Cd and non-metallic stressors such as desiccation and oxygen depletion. CdMT gene expression was upregulated by Cd exposure and desiccation, whereas no significant impact on the expression of CdMT was measured due to oxygen depletion. Overall, Cd was clearly more effective as an inducer of the CdMT gene expression compared to the applied non-metallic stressors. In unexposed snails, diurnal rhythmicity of CdMT gene expression was observed with higher mRNA concentrations at night compared to daytime. This rhythmicity was severely disrupted in Cd-exposed snails which exhibited highest CdMT gene transcription rates in the morning. Apart from diurnal rhythmicity, feeding activity also had a strong impact on CdMT gene expression. Although underlying mechanisms are not completely understood, it is clear that factors increasing MT expression variability have to be considered when using MT mRNA quantification as a biomarker for environmental stressors. [ABSTRACT FROM AUTHOR]

Anurans living in arid and semi-arid habitats are subjected to unpredictable rain patterns. Consequently, they should be prepared to reproduce at the onset of rain events. We investigated the covariation between calling behavior, testicular maturation, abdominal fat body index (FBI), plasma levels of androgens (T-DHT) and corticosterone (CORT) of males from three species of anurans in the Brazilian semi-arid during the reproductive period and drought. One of these species aestivates during the drought, while the other two species remain foraging. Although the three species display different behavioral strategies during the dry period, they present the same general reproductive patterns. T-DHT levels on the plasma and germinative cyst diameters were higher during the reproductive and breeding period compared to the drought. Additionally, the germinative cysts had all cell stages including sperm bundles during the dry season, however, it was only during the breeding event that free spermatozoa were found in the cyst lumen. These results suggest that these species present the reproductive pattern typical of desert anurans, consisting of opportunistic breeders that reproduce when triggered by a rain stimulus. Rhinella jimi and Pleurodema diplolister had higher CORT when males were calling. Moreover, Rhinella granulosa and P. diplolister showed lower FBI during breeding event, when males were calling. The high levels of CORT and lower FBI during reproductive period are associated, indicating that CORT modulates the recruitment of energy stores to prepare and maintain reproduction, particularly the expensive calling effort. [ABSTRACT FROM AUTHOR]

The sea cucumber Apostichopus japonicus is becoming an excellent model marine invertebrate for studies of environmentally-induced aestivation. Reversible protein phosphorylation as a regulatory mechanism in aestivation is known for some terrestrial aestivators but has never before been documented in sea cucumbers. The present study provides a global quantitative analysis of the role of reversible phosphorylation in sea cucumber aestivation by using tandem mass tag (TMT) labeling followed by an IMAC enrichment strategy to map aestivation-responsive changes in the phosphoproteome of sea cucumber intestine. We identified 2295 unique phosphosites derived from 1283 phosphoproteins and, of these, 211 hyperphosphorylated and 65 hypophosphorylated phosphoproteins were identified in intestine during deep aestivation compared with the active state based on the following criterion: quantitative ratios over 1.5 or less than 0.67 with corrected p-value < 0.05. Six major functional classes of proteins exhibited changes in their phosphorylation status during aestivation: (1) protein synthesis, (2) transcriptional regulators, (3) kinases, (4) signaling, (5) transporter, (6) DNA binding. These data on the global involvement of phosphorylation in sea cucumber aestivation significantly improve our understanding of the regulatory mechanisms involved in metabolic arrest when marine invertebrates face environmental stress and provide substantial candidate phosphorylated proteins that could be important for identifying functionally adaptive variation in marine invertebrates. Significance Sea cucumber Apostichopus japonicus is an excellent model organism for studies of environmentally-induced aestivation by a marine invertebrate. The present study provides the first quantitative phosphoproteomic analysis of sea cucumber aestivation using isobaric tag based TMT labeling followed by an IMAC enrichment strategy. These data on the global involvement of phosphorylation in sea cucumber aestivation significantly improve our understanding of the regulatory mechanism involved in metabolic arrest when marine invertebrates face environmental stress and provide substantial candidate phosphorylated proteins that could be important for identifying functionally adaptive variation in marine invertebrates. This study also demonstrates the usefulness of the TMT-based quantitative phosphoproteomics approach to explore the survival responses of a non-model marine invertebrate species to seasonal changes in its environment. [ABSTRACT FROM AUTHOR]

Progressive aridification since the mid-Miocene has had a significant influence on the evolution of the biota in the arid zone of central Australia. Especially moisture sensitive groups, such as snails, are often restricted to topographically complex areas, which have acted as refugia in an otherwise inhospitable environment. This historical fragmentation is deemed to be a potent agent of allopatric lineage diversification. Camaenid land snails are amongst only a few terrestrial gastropods that have managed to survive in the arid zone probably due to their ability to escape desiccation through aestivation. Here, we present the first study of the mitochondrial lineage differentiation in an endemic land snail genus from the Australian 'Red Centre', Granulomelon Iredale, 1933. Exposing significant incongruence between mtDNA phylogeny and morphology-based taxonomy, we completely revise the species and genus level taxonomy of this camaenid group. We demonstrate that this genus contains three species, G. grandituberculatum, G. adcockianum and G. squamulosum, which have so far been assigned to different genera: Granulomelon Iredale, 1933 (junior synonym: Baccalena Iredale, 1937), Basedowena Iredale, 1937 and Pleuroxia Ancey, 1887. Two of these species are widespread comprising multiple divergent mitochondrial lineages. Based on a molecular clock estimate, these lineages diverged approximately during the mid-Pleistocene, a period of particularly severe aridification. The phylogeographic patterns are consistent with an isolation-by-distance model in one species but not the other. We suggest that these differences can be attributed to their distinctive aestivation behavior. [ABSTRACT FROM AUTHOR]

A novel experimental approach for the simultaneous investigation of the effects of photoperiod and temperature showed evidence of diapause inHippodamia variegataGoeze but not inMicromus tasmaniaeWalker. Results are discussed in relation to their potential as greenhouse biocontrol agents. [ABSTRACT FROM AUTHOR]