Cuticle Hardness Research Articles

A comparison between sexes, castes, and life-histories supports the adaptive role of metal enrichment in the mandibles of Lasius ants

Ant mandibles perform different functions (e.g. feeding, hunting, and nest building), and previous studies on workers of some species revealed that they are enriched in transition metals (Zn and Mn), which improve cuticle hardness. Whether this trait is adaptive in all ant species is still unclear. One way to test for such adaptation is to compare sexes and castes within species as well as species with different life-histories. Here, we evaluated the transition metal enrichment in workers, queens, and males of 12 species of the Holarctic genus Lasius Fabricius 1804. The genus is adequate to test for the evolution of this trait because of two reasons. First, males show small, often toothless mandibles and do not contribute to nest activities, hence leading to the prediction of lower metal-enrichment compared with workers and queens. Second, this genus includes both socially parasitic (of other Lasius species) and non–parasitic species, leading to the prediction that females of the former, because they engage in fights with the host, have higher metal-enrichment compared with females of the latter. Our Scanning Electron Microscopy/Energy Dispersive X-ray analysis provided evidence for an adaptive role of Zn and Mn in Lasius mandibles. At intra-specific-level, males had lower amount of Zn and Mn in their mandibles compared with workers and queens (which did not differ). Although parasitic and non-parasitic queens had similar metal amounts, parasitic workers showed higher Mn content and marginally higher Zn content than workers of non-parasitic species. Overall, our results strongly suggest that both colony activities and parasitic life-style promoted greater metal-enrichment in Lasius females. Given the huge biological diversity of ants, large comparative studies are needed to assess the generality of our findings.

Cutting mechanics of wood by beetle larval mandibles

Wood boring is a feature of several insect species and is a major cause of severe and irreparable damage to trees. Adult females typically deposit their eggs on the stem surface under bark scales. The emerging hatchlings live within wood during their larval phase, avoiding possible predation, whilst continually boring and tunneling through wood until pupation. A study of wood boring by insects offers unique insights into the bioengineering principles that drive evolutionary adaptations. We show that larval mandibles of the coffee wood stem borer beetle (Xylotrechus quadripes: Cerambycidae) have a highly sharp cusp edge to initiate fractures in Arabica wood and a suitable shape to generate small wood chips that are suitable for digestion. Cuticle hardness at the tip is significantly enhanced through zinc-enrichment. A hollow architecture significantly reduces bending stresses at the mandibular base without compromising the structural integrity. Finite element model of the mandible showed highest stresses in the tip region; these decreased to significantly lower values at the start of the hollow section. A scaling model based on a fracture mechanics framework shows the importance of the mandible shape in generating optimal chip sizes. These findings contain general principles in tool design and put in focus interactions of insects and their woody hosts.

Supplementary Nutrition of Eucryptorrhynchus brandti (Coleoptera: Curculionidae: Cryptorrhychinae): Effect of Ailanthus altissima Host Tissues on Ovary Maturation and Oviposition.

Eucryptorrhynchus brandti is a trunk-boring pest that causes severe damage to its single host, Ailanthus altissima in China. Eucryptorrhynchus brandti adults require supplementary nutrition to achieve sexual maturity. Adults feed on petioles, shoots, 2- to 3-yr-old branches, and stems of A. altissima, but prefer to feed on stems. The effects of different host plant tissues on ovary development and oviposition in E. brandti are unclear. Thus, we examined the effects of the consumption of A. altissima petioles, shoots, branches, and stems on ovary development of E. brandti. Using ovaries of different ages, we also evaluated the differentiation of ovarioles, presence and appearance of follicular relics, number of eggs, cuticle hardness, and fat body quantity over time. We found the reproductive system was composed of two ovaries, lateral oviducts, a common oviduct, and spermatheca. The physiological age of ovaries was divided into three nulliparous and three parous stages. Females fed on stems had mature ovaries and successfully oviposited; females fed on other tree tissues or water had no functional ovaries and did not lay eggs. We found virgin females must obtain sufficient nutrition, as evidenced by the immature ovaries in females fed the water. We also found that only stems of A. altissima contribute to egg development; however, the other tree tissues do not contribute to egg development. These findings improve our understanding of the feeding preference of E. brandti from the perspective of ovary development as well as optimal host tissues for ovary development.

Age-dependent male mating tactics in a spider mite-A life-history perspective.

Males often fight with rival males for access to females. However, some males display nonfighting tactics such as sneaking, satellite behavior, or female mimicking. When these mating tactics comprise a conditional strategy, they are often thought to be explained by resource holding potential (RHP), that is, nonfighting tactics are displayed by less competitive males who are more likely to lose a fight. The alternative mating tactics, however, can also be explained by life‐history theory, which predicts that young males avoid fighting, regardless of their RHP, if it pays off to wait for future reproduction. Here, we test whether the sneaking tactic displayed by young males of the two‐spotted spider mite can be explained by life‐history theory. We tested whether young sneaker males survive longer than young fighter males after a bout of mild or strong competition with old fighter males. We also investigated whether old males have a more protective outer skin—a possible proxy for RHP—by measuring cuticle hardness and elasticity using nanoindentation. We found that young sneaker males survived longer than young fighter males after mild male competition. This difference was not found after strong male competition, which suggests that induction of sneaking tactic is affected by male density. Hardness and elasticity of the skin did not vary with male age. Given that earlier work could also not detect morphometric differences between fighter and sneaker males, we conclude that there is no apparent increase in RHP with age in the mite and age‐dependent male mating tactics in the mite can be explained only by life‐history theory. Because it is likely that fighting incurs a survival cost, age‐dependent alternative mating tactics may be explained by life‐history theory in many species when reproduction of old males is a significant factor in fitness.

Uncovering Ultrastructural Defences in Daphnia magna – An Interdisciplinary Approach to Assess the Predator-Induced Fortification of the Carapace

The development of structural defences, such as the fortification of shells or exoskeletons, is a widespread strategy to reduce predator attack efficiency. In unpredictable environments these defences may be more pronounced in the presence of a predator. The cladoceran Daphnia magna (Crustacea: Branchiopoda: Cladocera) has been shown to develop a bulky morphotype as an effective inducible morphological defence against the predatory tadpole shrimp Triopscancriformis (Crustacea: Branchiopoda: Notostraca). Mediated by kairomones, the daphnids express an increased body length, width and an elongated tail spine. Here we examined whether these large scale morphological defences are accompanied by additional ultrastructural defences, i.e. a fortification of the exoskeleton. We employed atomic force microscopy (AFM) based nanoindentation experiments to assess the cuticle hardness along with tapping mode AFM imaging to visualise the surface morphology for predator exposed and non-predator exposed daphnids. We used semi-thin sections of the carapace to measure the cuticle thickness, and finally, we used fluorescence microscopy to analyse the diameter of the pillars connecting the two carapace layers. We found that D . magna indeed expresses ultrastructural defences against Triops predation. The cuticle in predator exposed individuals is approximately five times harder and two times thicker than in control daphnids. Moreover, the pillar diameter is significantly increased in predator exposed daphnids. These predator-cue induced changes in the carapace architecture should provide effective protection against being crushed by the predator’s mouthparts and may add to the protective effect of bulkiness. This study highlights the potential of interdisciplinary studies to uncover new and relevant aspects even in extensively studied fields of research.

New Functions of Arthropod Bursicon: Inducing Deposition and Thickening of New Cuticle and Hemocyte Granulation in the Blue Crab, Callinectes sapidus

Arthropod growth requires molt-associated changes in softness and stiffness of the cuticle that protects from desiccation, infection and injury. Cuticle hardening in insects depends on the blood-borne hormone, bursicon (Burs), although it has never been determined in hemolymph. Whilst also having Burs, decapod crustaceans reiterate molting many more times during their longer life span and are encased in a calcified exoskeleton, which after molting undergoes similar initial cuticle hardening processes as in insects. We investigated the role of homologous crustacean Burs in cuticular changes and growth in the blue crab, Callinectes sapidus. We found dramatic increases in size and number of Burs cells during development in paired thoracic ganglion complex (TGC) neurons with pericardial organs (POs) as neurohemal release sites. A skewed expression of Burs β/Burs α mRNA in TGC corresponds to protein contents of identified Burs β homodimer and Burs heterodimer in POs. In hemolymph, Burs is consistently present at ∼21 pM throughout the molt cycle, showing a peak of ∼89 pM at ecdysis. Since initial cuticle hardness determines the degree of molt-associated somatic increment (MSI), we applied recombinant Burs in vitro to cuticle explants of late premolt or early ecdysis. Burs stimulates cuticle thickening and granulation of hemocytes. These findings demonstrate novel cuticle-associated functions of Burs during molting, while the unambiguous and constant presence of Burs in cells and hemolymph throughout the molt cycle and life stages may implicate further functions of its homo- and heterodimer hormone isoforms in immunoprotective defense systems of arthropods.

The movement and distribution of Helicoverpa armigera (Hübner) larvae on pea plants is affected by egg placement and flowering.

The distribution and movement of 1st instar Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae on whole garden pea (Pisum sativum L.) plants were determined in glasshouse trials. This economically-important herbivore attacks a wide variety of agricultural, horticultural and indigenous plants. To investigate the mechanisms underlying larval intra-plant movement, we used early-flowering and wild-type plant genotypes and placed eggs at different vertical heights within the plants, one egg per plant. Leaf water and nitrogen content and cuticle hardness were measured at the different plant heights. Of 92 individual larvae, 41% did not move from the node of eclosion, 49% moved upwards and 10% moved downwards with the distance moved being between zero and ten plant nodes. Larvae from eggs placed on the lower third of the plant left the natal leaf more often and moved further than larvae from eggs placed in the middle or upper thirds. The low nutritive value of leaves was the most likely explanation for more movement away from lower plant regions. Although larvae on flowering plants did not move further up or down than larvae on non-flowering plants, they more often departed the leaflet (within a leaf) where they eclosed. The final distribution of larvae was affected by plant genotype, with larvae on flowering plants found less often on leaflets and more often on stipules, tendrils and reproductive structures. Understanding intra-plant movement by herbivorous insects under natural conditions is important because such movement determines the value of economic loss to host crops. Knowing the behaviour underlying the spatial distribution of herbivores on plants will assist us to interpret field data and should lead to better informed pest management decisions.

Stiff coatings on compliant biofibers: the cuticle of Mytilus californianus byssal threads.

For lasting holdfast attachment, the mussel Mytilus californianus coats its byssal threads with a protective cuticle 2-5 microm thick that is 4-6 times stiffer than the underlying collagen fibers. Although cuticle hardness (0.1 GPa) and stiffness (2 GPa) resemble those observed in related mussels, a more effective dispersion of microdamage enables M. californianus byssal threads to sustain strains to almost 120% before cuticle rupture occurs. Underlying factors for the superior damage tolerance of the byssal cuticle were explored in its microarchitecture and in the cuticular protein, mcfp-1. Cuticle microstructure was distinctly granular, with granule diameters (approximately 200 nm) only a quarter of those in M. galloprovincialis cuticle, for example. Compared with homologous proteins in related mussel species, mcfp-1 from M. californianus had a similar mass (approximately 92 kDa) and number of tandemly repeated decapeptides, and contained the same post-translational modifications, namely, trans-4-hydroxyproline, trans-2,3-cis-3,4-dihydroxyproline, and 3,4-dihydroxyphenylalanine (Dopa). The prominence of isoleucine in mcfp-1, however, distinguished it from homologues in other species. The complete protein sequence deduced from cDNAs for two related variants revealed a highly conserved consensus decapeptide PKISYPPTYK that is repeated 64 times and differs slightly from the consensus peptide (AKPSYPPTYK) of both M. galloprovincialis and M. edulis proteins.

Metals and the Integrity of a Biological Coating: The Cuticle of Mussel Byssus

The cuticle of mussel byssal threads is a robust natural coating that combines high extensibility with high stiffness and hardness. In this study, fluorescence microscopy and elemental analysis were exploited to show that the 3,4-dihydroxyphenyl-L-alanine (dopa) residues of mussel foot protein-1 colocalize with Fe and Ca distributions in the cuticle of Mytilus galloprovincialis mussel byssal threads. Chelated removal of Fe and Ca from the cuticle of intact threads resulted in a 50% reduction in cuticle hardness, and thin sections subjected to the same treatment showed a disruption of cuticle integrity. Dopa-metal complexes may provide significant interactions for the integrity of composite cuticles deformed under tension.

Nano-Mechanical Investigation of the Byssal Cuticle, a Protective Coating of a Bio-Elastomer

ABSTRACTThe mechanical properties of the mussel byssal thread have been investigated via nano-indentation, with the emphasis on the differences between the cuticle and the fibrous interior. The cuticle hardness was found to be 30–40% higher than that of the underlying fibrous interior. In contrast, the Young's moduli in the two regions were virtually identical to one another. Elemental analysis via energy dispersive spectroscopy indicated surprisingly high levels of Al and Br in the cuticle considering the low amounts found in seawater. A potential role of Al in byssal thread mechanics is discussed in light of the unique capability of the cuticle to accommodate strains of 70% by the underlying fibrils in the core without delamination.

Nano-Mechanical Investigation of the Byssal Cuticle, a Protective Coating of a BioElastomer

ABSTRACTThe mechanical properties of the mussel byssal thread have been investigated via nano-indentation, with the emphasis on the differences between the cuticle and the fibrous interior. The cuticle hardness was found to be 30–40% higher than that of the underlying fibrous interior. In contrast, the Young's moduli in the two regions were virtually identical to one another. Elemental analysis via energy dispersive spectroscopy indicated surprisingly high levels of Al and Br in the cuticle considering the low amounts found in seawater. A potential role of Al in byssal thread mechanics is discussed in light of the unique capability of the cuticle to accommodate strains of 70% by the underlying fibrils in the core without delamination.

Diflubenzuron Effects on Boll Weevils (Coleoptera: Curculionidae) in Small Field Cages

In 1992 and 1993, newly emerged adult boll weevils, Anthonomus grandis grandis Boheman, were released on diflubenzuron (DFB)-treated and untreated cotton plants grown in 20 screened field cages. Seven days later weevils were recovered, and the effects of DFB on adult survival, mating success of males, egg hatch, flight activity, and cuticle hardness were measured. DFB substantially reduced hatch, flight activity, and cuticle hardness in both years; survival apparently was reduced in 1992, but not in 1993. Mating success of treated males was unaffected in either year. The combined effect of reduced egg hatch, flight activity, cuticle hardness, and possibly survival demonstrates several ways in which DFB may reduce populations of boll weevils in the field.

Diflubenzuron Plus Cottonseed Oil: Effects on Boll Weevil (Coleoptera: Curculionidae) Cuticle Hardness, Mating, and Flight

Laboratory studies were done to measure physiological effects of diflubenzuron when formulated in cottonseed oil and applied at very low rates to boll weevils, Anthonomus grandis grandis Boheman. Tests were designed to measure cuticle softness, mating propensity, and effect on flight. We found significant differences within treatments in all three studies as the dose increased. Cottonseed oil formulations containing 25% diflubenzuron reduced cuticle hardness by 46.2–61.0%. Cottonseed oil formulations containing 12.5–25% diflubenzuron reduced mating by 22.2–35.4% and flight by 11.6–21.2%.

On the Indentation Hardness of Insect Cuticle

ABSTRACT The indentation hardness of locust cuticle was measured using a Vickers diamond applied to the surface of the cuticle. A number of areas of cuticle showed similar hardness values, approximately 24 kg mm−2, for the dehydrated cuticle but the mandibles (29·8 kg mm−2) and the dorsal mesothorax (33·3 kg mm−2) were harder. The mandibles have localized areas of hardness which are related to their function. It is suggested that the hardness of the dorsal thorax may be related to its function as an energy storing structure in flight. The measured hardness of cuticle depends on the hydration of the material but relative differences between cuticles from different parts of the locust persist whether tested fresh or after dehydration. Hardness is correlated with the dihydroxyphenol content of cuticle (unlike stiffness), conversely it is not correlated with the hydrophobicity of the matrix proteins (unlike stiffness). It is emphasized that hardness and stiffness are very different mechanical properties.

Spider Populations in Alfalfa, with Notes on Spider Prey and Effect of Harvest

The spider fauna of a Virginia alfalfa field was determined, and the seasonal occurrence of the 4 major species was plotted. Samples were taken with a D-Vac® sampler and a sweep net. Fourteen families were represented and comprised 75 genera and 112 identifiable species. Tetragnatha laboriosa Hentz, Pachygnatha tristriata Koch, Misumeuops asperatus (Hentz), and Oxyopes salticus Hentz were the most common. The effect of alfalfa harvest on numbers of spiders in the field was measured. The numbers of T. laboriosa, P. tristriata , and the family Lycosidae were not affected by the cutting process, but those of the Linyphiidae, Thomisidae, and Salticidae showed a definite response. Collections at intervals, over 24-hour periods, showed the effect of spider activity on sampling results. P. tristriata was far more active on the plant at night, and this was believed to be the result of a negative photo-orientation. T. laboriosa was collected during the periods of highest temperatures and lowest humidities. Feeding tests with 10 species belonging to 5 families were conducted in the laboratory. Although other factors were apparent, it appeared that size of prey, speed, and hardness of cuticle were the major criteria for determining whether a spider would feed on a certain insect.