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Concept: Ecdysis

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A nautiloid conch containing many disarticulated exoskeletons of Omegops cornelius (Phacopidae, Trilobita) was found in the Upper Devonian Hongguleleng Formation of the northwestern margin of the Junggar Basin, NW China. The similar number of cephala, thoraces and pygidia, unbroken thoraces, explicit exuviae, and lack of other macrofossils in the conch, indicate that at least seven individual trilobites had moulted within the nautiloid living chamber, using the vacant chamber of a dead nautiloid as a communal place for ecdysis. This exuvial strategy manifests cryptic behaviour of trilobites, which may have resulted from the adaptive evolution induced by powerful predation pressure, unstable marine environments, and competition pressure of organisms occupying the same ecological niche in the Devonian period. The unusual presence of several trilobites moulting within a nautiloid conch is possibly associated with social behaviours in face of a serious crisis. New materials in this study open a window for understanding the survival strategy of marine benthic organisms, especially predator-prey interactions and the behavioural ecology of trilobites in the middle Palaeozoic.

Concepts: Arthropod, Predation, Ecology, Carboniferous, Ordovician, Trilobite, Devonian, Ecdysis

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Organisms in the wild have cryptic life stages that are sensitive to changing environmental conditions and can be difficult to survey. In this study, I used mark-recapture methods to repeatedly survey Anaea aidea (Nymphalidae) caterpillars in nature, then modeled caterpillar demography as a hidden Markov process to assess if temporal variability in temperature and density influence the survival and growth of A. aidea over time. Individual encounter histories result from the joint likelihood of being alive and observed in a particular stage, and I have included hidden states by separating demography and observations into parallel and independent processes. I constructed a demographic matrix containing the probabilities of all possible fates for each stage, including hidden states, e.g., eggs and pupae. I observed both dead and live caterpillars with high probability. Peak caterpillar abundance attracted multiple predators, and survival of fifth instars declined as per capita predation rate increased through spring. A time lag between predator and prey abundance was likely the cause of improved fifth instar survival estimated at high density. Growth rates showed an increase with temperature, but the preferred model did not include temperature. This work illustrates how state-space models can include unobservable stages and hidden state processes to evaluate how environmental factors influence vital rates of cryptic life stages in the wild.

Concepts: Predation, Ecology, Markov chain, Hidden Markov model, Lepidoptera, Caterpillar, Andrey Markov, Ecdysis

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Biological materials are the result of years of evolution and possess a number of efficient features and structures. Researchers have investigated the possibility of designing biomedical structures that take advantage of these structural features. Insect shells, such as beetle shells, are among the most promising types of biological material for biomimetic development. However, due to their intricate geometries and small sizes, it is challenging to measure the mechanical properties of these microscale structures. In this study, we developed an in-situ testing platform for site-specific experiments in a focused ion beam (FIB) system. Multi-axis nano-manipulators and a micro-force sensor were utilized in the testing platform to allow better results in the sample preparation and data acquisition. The entire test protocol, consisting of locating sample, ion beam milling and micro-mechanical bending tests, can be carried out without sample transfer or reattachment. We used our newly devised test platform to evaluate the micromechanical properties and structural features of each separated layer of the beetle horn shell. The Young’s modulus of both the exocuticle and endocuticle layers was measured. We carried out a bending test to characterize the layers mechanically. The exocuticle layer bent in a brick-like manner, while the endocuticle layer exhibited a crack blunting effect.

Concepts: Insect, Arthropod, Measurement, Young's modulus, Solid mechanics, Focused ion beam, Procuticle, Ecdysis

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Locusts jump by using a catapult mechanism in which energy produced by slow contractions of the extensor tibiae muscles of the hind legs is stored in distortions of the exoskeleton, most notably 1) the two semi-lunar processes at each knee joint and 2) the tendons of the extensor muscles themselves. The energy is then suddenly released from these two stores to power the rapid, propulsive movements of the hind legs. The reliance on the mechanical storage of energy is likely to impact on jumping because growth occurs by a series of five moults, at each of which the exoskeleton is replaced by a new one. All developmental stages (instars) nevertheless jump as a means of forward locomotion, or as an escape movement. Here I show that in each instar, resilin is added to the semi-lunar processes and to the core of the extensor tendons so that their thickness increases. As the next moult approaches, a new exoskeleton forms within the old one with resilin already present in the new semi-lunar processes. The old exoskeleton, the tendons, and their resilin is discarded at moulting. The resilin of the semi-lunar processes and tendons of the new instar is initially thin, but a similar pattern of deposition results in an increase of their thickness. In adults, resilin continues to be deposited so that at four weeks old the thickness in the semi-lunar processes has increased fourfold. These changes in the energy stores accompany changes in jumping ability and performance during each moulting cycle.

Concepts: Insect, Developmental biology, Knee, Flexion, Extension, Semimembranosus muscle, Jumping, Ecdysis

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The polyhydroxylated derivatives of 6-keto,7-dehydrocholesterol (ecdysone, ecdysteroids, Ecd) are natural compounds widely distributed in plants. They exhibit strong anabolic, vitamin D-like, pharmacological effects in vertebrate animals and in the human body. In the larval stages of insects, injections of pure Ecd cause serious pathophysiological, “hyperecdysonic” syndromes associated with neuromuscular paralysis, premature cuticular apolysis and complete inhibition of ecdysis. Ecd do not penetrate insect cuticle. For this reason, all previous attempts to induce ecdysone responses by topical applications of Ecd failed. In this work, we tried to induce the topical effects of Ecd by preparation of more lipophilic complexes, with 2 or 4 molecules of 20-hydroxyecdysone (E20) attached to a relatively large nucleus of the porphyrin. The resulting porphyrin-E20 complexes (ecdysogens) have been subjected to standardized assays for ecdysone activity in the ligatured larvae (dauerlarvae) of the greater waxmoth (Galleria mellonella). Similarly like the free E20 alone, porphyrin-E20 complexes had no effect when applied on the body surface or administered in the larval diet. When injected, however, they exhibited delayed effects, but the adverse (“hyperecdysonic”) pathophysiological syndromes were reduced or abolished. It is concluded, therefore, that the replacement of pathophysiological, precocious or “hyperecdysonic” moults by the larval-pupal transformation, was due to successive metabolic liberation of the biologically active, free E20 from the porphyrin-E20 complex. The biological status of Ecd does not agree with their definition as the prothoracic gland (PG) hormone of insects, nor with the assumptions about a growth hormone of plants. A possibility that the most important status of Ecd may depend on the pharmacological properties of a sterolic D6 vitamin has been discussed.

Concepts: Metabolism, Insect, Developmental biology, Larva, Arthropod, Growth hormone, Human body, Ecdysis

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Adult females of Pyrrhocoris apterus, programmed for diapause by short-day (SD) photoperiod and those programmed for reproduction by long-day (LD) retain photoperiodic information in continuous darkness (DD) until death. However, if the interruption of SD by DD is made in the course of diapause programming in adults, then the incidence of diapause depends on the number of SD cycles received before DD, with no evidence that the photoperiodic clock is free-running at DD to complete diapause induction. These results indicate that the photoperiodic clock is stopped after transfer to DD and the information accumulated before transfer to DD is maintained. Diapause programming in the adult stage requires 9-10 SD cycles to induce diapause in 80% of individuals. However, if the diapause programming starts after ecdysis of LD-larvae to the last instar, only 3 SD cycles before transfer to DD are required for diapause in 80% of individuals. Surprisingly, if the newly ecdysed last instar LD-larvae, sensitive to photoperiod, are transferred to DD (thus they did not experience any SD), diapause occurs in 40% of the individuals. Thus, diapause ‘information’ is present in LD-larvae and is responsible for a lower number of SD required for diapause induction in the larval than in the adult stage.

Concepts: Time, Developmental biology, Hemiptera, Adult, Photoperiodism, Diapause, Firebug, Ecdysis

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The exoskeletons of pancrustaceans, as typified by decapod crustaceans and insects, demonstrate a high degree of similarity with respect to histology, ultrastructure, function, and composition. The cuticular envelope in insects and the outer epicuticle in crustaceans both serve as the primary barrier to permeability of the exoskeleton, preventing loss of water and ions to the external medium. Prior to and following ecdysis, there is a sequence of expression and synthesis of different proteins by the cuticular epithelium for incorporation into the pre-exuvial and post-exuvial procuticle of insects and the exocuticle and endocuticle of crustaceans. Both exhibit regional differences in cuticular composition, e.g., the articular (intersegmental) membranes of insects and the arthrodial (joint) membranes of crustaceans. The primary difference between these cuticles is the ability to mineralize. Crustaceans' cuticles express a unique suite of proteins that provide for the nucleation and deposition of calcium carbonate. Orthologs of genes discussed in the present review were mined from a recently completed cuticular transcriptome of the crayfish, Cherax quadricarinatus, providing new insights into the nature of these proteins.

Concepts: DNA, Insect, Arthropod, Crustacean, Exoskeleton, Epicuticle, Procuticle, Ecdysis

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The bag-shelter moth, Ochrogaster lunifer Herrich-Schaffer (Lepidoptera: Notodontidae), is associated with a condition called equine amnionitis and fetal loss (EAFL) on horse farms in Australia. Setal fragments from O. lunifer larvae have been identified in the placentas of experimentally aborted fetuses and their dams, and in clinical abortions. The gregarious larvae build silken nests in which large numbers cohabit over spring, summer and autumn. The final instars disperse to pupation sites in the ground where they overwinter. Field-collected O. lunifer larvae, their nests and nearby soil were examined using light and electron microscopy to identify setae likely to cause EAFL and to determine where and how many were present. Microtrichia, barbed hairs and true setae were found on the exoskeletons of the larvae. True setae matching the majority of setal fragments described from equine tissue were found on third to eighth instar larvae or exuviae. The number of true setae increased with the age of the larva; eighth instars carried around 2.0-2.5 million true setae. The exuvia of the pre-pupal instar was incorporated into the pupal chamber. The major sources of setae are likely to be nests, dispersing pre-pupal larvae and their exuviae, and pupal chambers.

Concepts: Pregnancy, Fetus, Insect, Developmental biology, Abortion, Lepidoptera, Caterpillar, Ecdysis

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This study examined the mesocardiac and urocardiac ossicles in the gastric mill of the blue crab to describe its structure, mineralization, and dynamics throughout the molt cycle, and to assess its possible utility in age determination. Morphologically, the mineralized ossicles are similar to the calcified dorsal carapace having a lamellate structure comprised of sheets of chitin/protein fibrils. Staining with acridine orange showed the same arrangement of an epicuticle, exocuticle, and endocuticle. In much of the mesocardiac and urocardiac ossicles, the endocuticle is very reduced, with the exocuticle predominating; the reverse of the dimensions of the exoskeleton. The lamellate structure of the ossicles was confirmed with scanning electron microscopy; however, elemental mapping by energy-dispersive analysis of X-rays revealed that the ossicles are mineralized with calcium phosphate, in contrast to the calcium carbonate biomineral of the exoskeleton. The medial tooth of the urocardiac ossicle is not calcified, but the epicuticle is highly elaborated and impregnated with silica. Histological examination of the ossicles demonstrated that they are molted during ecdysis, so despite the appearance of bands in the mesocardiac ossicle, it is difficult to hypothesize how the bands could represent a record of chronological age. J. Morphol. 276:1358-1367, 2015. © 2015 Wiley Periodicals, Inc.

Concepts: Insect, Arthropod, Blue crab, Crab, Callinectes, Portunidae, Epicuticle, Ecdysis

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We studied the response of gypsy moth (Lymantria dispar (Linnaeus) (Lepidoptera: Lymantriidae)) to the content of manganese in food in the laboratory breeding of caterpillars. The food of the caterpillars {Betula pendula Roth (Fagales: Betulaceae) leaves} was contaminated by dipping in the solution of MnCl2·4H2O with manganese concentrations of 0, 0.5, 5 and 10 mg ml(-1), by which differentiated manganese contents (307; 632; 4,087 and 8,124 mg kg(-1)) were reached. Parameters recorded during the rearing were as follows: effect of manganese on food consumption, mortality and length of the development of caterpillars, pupation and hatching of imagoes. At the same time, manganese concentrations were determined in the offered and unconsumed food, excrements, and exuviae of the caterpillars, pupal cases and imagoes by using the AAS method. As compared with the control, high manganese contents in the food of gypsy moth caterpillars affected the process of development particularly by increased mortality of the first instar caterpillars (8 % mortality for caterpillars with no Mn contamination (T0) and 62 % mortality for subjects with the highest contamination by manganese (T3)), by prolonged development of the first-third instar (18.7 days (T0) and 27.8 days (T3)) and by increased food consumption of the first-third instar {0.185 g of leaf dry matter (T0) and 0.483 g of leaf dry matter (T3)}. The main defence strategy of the caterpillars to prevent contamination by the increased manganese content in food is the translocation of manganese into frass and exuviae castoff in the process of ecdysis. In the process of development, the content of manganese was reduced by excretion in imagoes to 0.5 % of the intake level even at its maximum inputs in food.

Concepts: Insect, Developmental biology, Lepidoptera, Manganese, Pupa, Lymantriidae, Gypsy moth, Ecdysis