Larval Head Research Articles

Foraging in the darkness: Highly selective tuning of below‐ground larval olfaction to Brassicaceae volatiles in striped flea beetle

AbstractThe olfactory system of above‐ground insects is among the best described perceptual architectures. However, remarkably little is known about how below‐ground insects navigate in the dark for foraging. Here, we investigated host plant preferences, olfactory sensilla and characterise olfactory proteins in below‐ground larvae of the striped flea beetle (SFB) Phyllotreta striolata Fabricius (Coleoptera: Chrysomelidae). Both the adults and larvae of this coleopteran pest cause serious damage to Brassicaceous crops above and below ground, respectively. To elucidate the role of olfactory system in host location of below‐ground larvae, we initially demonstrated that SFB larvae distinctly favoured Brassicaceae over other plant families by two‐choice behavioural bioassay. Subsequently, scanning electron microscopy of sensilla in SFB larval head showed a significant reduction in the number of olfactory sensilla in larvae compared with adults. However, essential olfactory sensilla such as sensilla basiconica are underscoring the indispensability of the larval olfactory system. We selected four larval‐specific odorant binding proteins for functional validation from our previous transcriptome data. Functional studies revealed that PstrOBP23 exhibits robust binding affinity to 24 volatiles of Brassicaceae plants, including seven isothiocyanate compounds. This suggests a pivotal role of PstrOBP23 in the foraging behaviour of the larvae below the ground. Moreover, two ligands displaying strong binding capacity exhibit apparent attractive or repellent activity towards SFB larvae. Our findings provide a crucial insight into the olfactory system of below‐ground larvae in SFB, highlighting the highly selective tuning of larvae specific OBP to host plant volatiles. These results offer potential avenues for developing effective pest control strategies against SFB.

Cerebral Vascular Toxicity after Developmental Exposure to Arsenic (As) and Lead (Pb) Mixtures.

Arsenic (As) and lead (Pb) are environmental pollutants found in common sites linked to similar adverse health effects. This study determined driving factors of neurotoxicity on the developing cerebral vasculature with As and Pb mixture exposures. Cerebral vascular toxicity was evaluated at mixture concentrations of As and Pb representing human exposures levels (10 or 100 parts per billion; ppb; µg/L) in developing zebrafish by assessing behavior, morphology, and gene expression. In the visual motor response assay, hyperactivity was observed in all three outcomes in dark phases in larvae with exposure (1-120 h post fertilization, hpf) to 10 ppb As, 10 ppb Pb, or 10 ppb mix treatment. Time spent moving exhibited hyperactivity in dark phases for 100 ppb As and 100 ppb mix treatment groups only. A decreased brain length and ratio of brain length to total length in the 10 ppb mix group was measured with no alterations in other treatment groups or other endpoints (i.e., total larval length, head length, or head width). Alternatively, measurements of cerebral vasculature in the midbrain and cerebellum uncovered decreased total vascularization at 72 hpf in all treatment groups in the mesencephalon and in all treatment groups, except the 100 ppb Pb and 10 ppb As groups, in the cerebellum. In addition, decreased sprouting and branching occurred in the mesencephalon, while only decreased branching was measured in the cerebellum. The 10 ppb Pb group showed several cerebral vasculature modifications that were aligned with a specific gene expression alteration pattern different from other treatment groups. Additionally, the 100 ppb As group drove gene alterations, along with several other endpoints, for changes observed in the 100 ppb mix treatment group. Perturbations assessed in this study displayed non-linear concentration-responses, which are important to consider in environmental health outcomes for As and Pb neurotoxicity.

Unraveling the Role of Cuticular Protein 3-like (HvCP3L) in the Chitin Pathway through RNAi and Methoxyfenozide Stress Response in Heortia vitessoides Moore.

Cuticle proteins (CPs) constitute a multifunctional family; however, the physiological role of Cuticle Protein 3-like (CP3L) in Heortia vitessoides Moore remains largely unclear. In this study, we cloned the HvCP3L gene from the transcriptional library of Heortia vitessoides Moore. RT-qPCR results revealed that HvCP3L exhibited high expression levels during the larval stage of Heortia vitessoides Moore, particularly at the L5D1 stage, observed in both larval and adult heads. Through RNA interference, we successfully silenced the HvCP3L gene, resulting in a significant reduction in the survival rate of Heortia vitessoides Moore, with the survival rate from larvae to adults plummeting to a mere 17.7%, accompanied by phenotypic abnormalities. Additionally, we observed that the knockdown of HvCP3L led to the inhibition of genes in the chitin pathway. Following exposure to methoxyfenozide stress, the HvCP3L gene exhibited significant overexpression, coinciding with phenotypic abnormalities. These findings underscore the pivotal role of HvCP3L in the growth and development of Heortia vitessoides Moore.

Identification of odorant binding protein and chemosensory protein genes in Protegira songi (Lepidoptera Noctuidae) via transcriptome analysis

The larvae of Protegira songi Chen & Zhang are notorious defoliator specifically infesting the medicinal plant Eucommia ulmoides Oliver. In insects, Odorant binding proteins (OBPs) and chemosensory proteins (CSPs) are highly expressed in olfactory organs and play essential roles in perceiving female pheromone and host volatiles by binding and transporting odorant compounds. In this study, 17 OBPs and 16 CSPs were identified from P. songi by transcriptome analysis. Sequence alignment indicated that PsonOBPs and PsonCSPs share no identity with each other in protein sequence. The overall structures of PsonOBPs and PsonCSPs are mainly constructed by α-helix bundles and loops. But the distribution of conserved cysteine residues and arrangement of α-helix are varied between PsonOBPs and PsonCSPs. Putative ligand binding cavities were found in these predicted protein structures. Expression profiles of PsonOBPs and PsonCSPs in adults and larvae were determined by RT-PCR which showed that 16 PsonOBPs and 11 PsonCSPs were expressed in adult antennae or/and larval head. PsonGOBPs, PsonPBPs, PsonOBP5, PsonOBP7, and PsonCSP6 were preferentially expressed in olfactory associated tissues. Our results suggest that the PsonOBPs and PsonCSPs that contain ligand cavity and are highly enriched in olfactory tissues may play roles in chemosensory perception of P. songi, and the transcriptome data may facilitate the studies on the olfactory mechanism of P. songi specifically choosing E. ulmoides as host.

Transcriptome-wide analysis uncovers regulatory elements of the antennal transcriptome repertoire of bumblebee at different life stages.

Bumblebees are crucial pollinators, providing essential ecosystem services and global food production. The success of pollination services relies on the interaction between sensory organs and the environment. The antenna functions as a versatile multi-sensory organ, pivotal in mediating chemosensory/olfactory information, and governs adaptive responses to environmental changes. Despite an increasing number of RNA-sequencing studies on insect antenna, comprehensive antennal transcriptome studies at the different life stages were not elucidated systematically. Here, we quantified the expression profile and dynamics of coding/microRNA genes of larval head and antennal tissues from early- and late-stage pupa to the adult of Bombus terrestris as suitable model organism among pollinators. We further performed Pearson correlation analyses on the gene expression profiles of the antennal transcriptome from larval head tissue to adult stages, exploring both positive and negative expression trends. The positively correlated coding genes were primarily enriched in sensory perception of chemical stimuli, ion transport, transmembrane transport processes and olfactory receptor activity. Negatively correlated genes were mainly enriched in organic substance biosynthesis and regulatory mechanisms underlying larval body patterning and the formation of juvenile antennal structures. As post-transcriptional regulators, miR-1000-5p, miR-13b-3p, miR-263-5p and miR-252-5p showed positive correlations, whereas miR-315-5p, miR-92b-3p, miR-137-3p, miR-11-3p and miR-10-3p exhibited negative correlations in antennal tissue. Notably, based on the inverse expression relationship, positively and negatively correlated microRNA (miRNA)-mRNA target pairs revealed that differentially expressed miRNAs predictively targeted genes involved in antennal development, shaping antennal structures and regulating antenna-specific functions. Our data serve as a foundation for understanding stage-specific antennal transcriptomes and large-scale comparative analysis of transcriptomes in different insects.

Descriptions of the larvae and pupae of Sisko Ross with notes on western Dolophilodes Ulmer larvae (Trichoptera: Philopotamidae)

Larvae and a pupa of the caddisfly genus Sisko Ross (Trichoptera: Philopotamidae) are described. Larval and pupal photos and larval diagnoses are provided. Larval head and mandible photos and notes are provided for four species of western Dolophilodes Ulmer. A key to the larvae of North American Philopotamidae genera is included.

The families of the suborder Brachycera Schiner, 1862 (Insecta: Diptera): Part 2

Most Diptera belong to the Brachycera, a group characterized by the reduction or fusion of antennal segments to eight or fewer and by modifications to the larval head and mouthparts. The larvae of most species are predatory, although there are a few parasitoid groups as well. From a methodological point of view, the research was conducted using bibliometric and content analysis approaches. Bibliometrics is guided by the principle of analyzing scientific or technical activity through quantitative studies of publications, that is, through this approach. Document analysis was used as a data collection method to gather information from theoretical books, banks, university dissertations, scientific journals, documents, and digital platforms.

Influence of short-term, water-deprived bermudagrass on Orius insidiosus predation and Spodoptera frugiperda larval survival and development

ABSTRACT Turfgrass is susceptible to changes in water availability, and its effects could cascade through multiple trophic levels. In some years, turfgrass is exposed to short drought spells. It is unclear if this short period of water deprivation would cascade to predator performance on herbivory on bermudagrass. Thus, the objective of this study was to determine the effects of water deprivation on predation by Orius insidiosus (Say) (Hemiptera: Anthocoridae) on Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) when mediated by drought-tolerant ‘TifTuf’ and drought-susceptible ‘Tifway’ bermudagrass. Neonates of S. frugiperda and adults of O. insidiosus were caged on bermudagrass in greenhouse conditions. Treatments were three factors: 1) water deprivation at three levels – fully, moderately, and sparsely watered; 2) predator at two levels – present or absent; and 3) bermudagrass at two levels – drought tolerant and susceptible cultivars. The survival of S. frugiperda larvae was reduced when O. insidiosus adults were present regardless of water deprivation levels and bermudagrass cultivars. Reduced larval weight, body length, and head capsule width of S. frugiperda larvae were observed with predators compared to without predators. S. frugiperda larval mortality was significantly greater with predator’s absence for the sparsely-watered treatment than for the fully and moderately-watered treatments. The levels of water deprivation had a partial impact on the survival of S. frugiperda larvae, whereas bermudagrass cultivars did not affect the survival and development of S. frugiperda larvae. Thus, during at least short-term dry spells, O. insidiosus predation can potentially reduce the S. frugiperda population on bermudagrass.

Application of noninvasive sampling technique in mitochondrial genome intraspecific phylogeny of the endangered butterfly, Teinopalpus aureus (Lepidoptera: Papilionidae).

The butterfly genus of Teinopalpus, endemic to Asia, embodies a distinct species of mountain-dwelling butterflies with specific habitat requirements. These species are rare in the wild and hold high conservation and research value. Similar to other protected species, the genetic analysis of the rare Teinopalpus aureus poses challenges due to the complexity of sampling. In this study, we successfully extracted DNA and amplified mitochondrial genomic DNA from various noninvasive sources such as larval feces, larval exuviae, larval head capsules, pupal exuviaes, and filamentous gland secretions, all integral parts of butterfly metamorphosis. This was conducted as part of a research initiative focused on the artificial conservation of T. aureus population in Jinggang Shan Nature Reserve. Our findings illustrated the successful extraction of DNA from multiple noninvasive sources, achieved through modified DNA extraction methodologies. Although the DNA concentration obtained from noninvasive samples was lower than that from muscle tissues of newly dead larvae during rearing, all samples met the requirements for PCR amplification and sequencing, yielding complete circular sequences. These sequences are pivotal for both interspecific and intraspecific genetic relationship analysis. Our methods can be extended to other insects, especially scarce species.

Contribution to the knowledge of larval chaetotaxy of the genus Enochrus Thomson: description of five New World species (Coleoptera: Hydrophilidae)

The primary and secondary chaetotaxy of the larval head capsule and head appendages of five New World species of Enochrus Thomson are described for the first time. These species belong to three subgenera: Enochrus (Hugoscottia) tremolerasi (Knisch), E. (Lumetus) hamiltoni (Horn), E. (Methydrus) barituensis Fernández, E. (Methydrus) ochraceus (Melsheimer) and E. (Methydrus) pygmaeus (Fabricius). Morphometric characters derived from the head capsule and mouthparts are also included, together with detailed illustrations of all characters. A comparative study of the chaetotaxy of four subgenera of Enochrus yields an important number of characters useful to diagnose the larvae of these subgenera, and could be used in future phylogenetic analyses. Several characters showing variation within a subgenus are discussed; additionally, possible diagnostic characters for Enochrus and the subfamily Enochrinae are discussed. The chaetotaxy of two subgenera, Enochrus Thomson and Hydatotrephis MacLeay, still remains unknown, and should be a priority to complete the larval knowledge of the genus Enochrus.

Molecule characterization of chemosensory and metabolism-related genes in the proboscis of Athetis lepigone.

Introduction: The moth species Athetis lepigone (Möschler) (Lepidoptera: Noctuidae), which has recently been identified as a pest of summer maize (Zea mays L.) in China, has demonstrated a rapid proliferation with in the Huang-Huai-Hai Plain region since its initial discovery in Hebei Province in 2005. It has become a prevalent pest of corn crops, and its ability to adapt quickly to its surroundings is currently being investigated. One of the key characteristics of its siphoning mouthparts is not only the feeding apparatus itself but also the chemosensory organs that enable the detection of chemical signals from the surrounding environment. However, there is a lack of comprehensive research on the genes responsible for chemosensory and metabolic mechanisms in the proboscises of male and female A. lepigone adults. Methods: In this study, we utilized transcriptome analysis to identify a total of fifty chemosensory genes from six distinct families, including 19 odorant-binding proteins (OBPs), 22 chemosensory proteins (CSPs), one co-receptor (Orco), six odorant receptors (ORs), four ionotropic receptors (IRs), and two sensory neuron membrane proteins (SNMPs) in the proboscis. Notably, seven OBPs, two CSPs, and one OR were discovered for the first time. Additionally, fourteen genes related to metabolism, including cytochrome P450 (CYPs) and carboxylesterases (CXEs), were also identified. Furthermore, a qualitative analysis was conducted on the relative transcript levels of eight related genes. The expression of 21 annotated chemosensory and metabolic genes was compared between A. lepigone adults and larvae using qRT-PCR, revealing tissue specificity. The majority of genes exhibited predominant expression in the antennae and proboscis during the adult stage, while showing slight expression in the combination of sixth-instar larval head oral appendages (maxilla, labium, and antenna) and pheromone gland-ovipositors of female adults. Results/discussion: Our study points to a new pest control strategies that these newly discovered genes have the potential to serve as targets for enhancing future pest control, including mating disruption and the use of food attractants. And it would be advantageous to ascertain the distribution of chemosensory gene expression and gain insights into the functionalities of these genes, thereby establishing a novel theoretical framework for the advancement of eco-friendly pesticides and efficient pest management strategies in the future.

Comparative Transcriptomic Assessment of Chemosensory Genes in Adult and Larval Olfactory Organs of Cnaphalocrocis medinalis.

The rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae), is a notorious pest of rice in Asia. The larvae and adults of C. medinalis utilize specialized chemosensory systems to adapt to different environmental odors and physiological behaviors. However, the differences in chemosensory genes between the olfactory organs of these two different developmental stages remain unclear. Here, we conducted a transcriptome analysis of larvae heads, male antennae, and female antennae in C. medinalis and identified 131 putative chemosensory genes, including 32 OBPs (8 novel OBPs), 23 CSPs (2 novel CSPs), 55 ORs (17 novel ORs), 19 IRs (5 novel IRs) and 2 SNMPs. Comparisons between larvae and adults of C. medinalis by transcriptome and RT-qPCR analysis revealed that the number and expression of chemosensory genes in larval heads were less than that of adult antennae. Only 17 chemosensory genes (7 OBPs and 10 CSPs) were specifically or preferentially expressed in the larval heads, while a total of 101 chemosensory genes (21 OBPs, 9 CSPs, 51 ORs, 18 IRs, and 2 SNMPs) were specifically or preferentially expressed in adult antennae. Our study found differences in chemosensory gene expression between larvae and adults, suggesting their specialized functions at different developmental stages of C. medinalis. These results provide a theoretical basis for screening chemosensory genes as potential molecular targets and developing novel management strategies to control C. medinalis.

Morphometrics of <i>Spodoptera frugiperda</i> (J E Smith) as Influenced by Cereal Hosts

This laboratory study evaluate the morphometrics of fall army worm Spodoptera frugiperda (J E Smith) influenced by four cereal host plants. In maize, larval head capsule width was 0.22, 0.42, 0.71, 1.13, 1.29 and 2.94 mm for I to VI instars, respectively in sorghum was 0.20, 0.41, 0.67, 1.06, 1.24 and 3.05 mm for I to VI instars, respectively. In pearl millet, larval head capsule width was 0.19, 0.29, 0.52, 0.83, 1.23 and 2.80 mm, for I to VI instars, respectively. In sugarcane, head capsule width was 0.22, 0.43, 0.67, 0.94, 1.22 and 2.70 mm for I to VI instars, respectively. The mean pupal length varied significantly when reared on different cereal host plants, it was significantly maximum on maize (15.10 mm) followed by sorghum (14.20 mm), sugarcane (13.10) and lowest on pearl millet (12.30 mm). The significantly maximum pupal weight was noticed on maize (185.90 mg) followed by sorghum (144.40 mg), sugarcane (124.80 mg) and lowest on pearl millet (104.70 mg).

Morphometrics of <i>Pectinophora gossypiella</i> (Saunders) on Cotton as Influenced by Temperature

The morphometry of life stages of the pink boll worm Pectinophora gossypiella (Saunders) was evaluated at four temperature levels of 20, 25, 30 and 35°C to assess the impact of temperature. Measurement of larval head capsule width, body length, body width and weight on cotton at 20°C were 0.68, 0.87, 0.98 and 1.24 mm, 2.27, 5.80, 7.46 and 10.60 mm, 1.08, 1.58, 1.68 and 2.53 mm and 0.40, 2.84, 24.42 and 64.21 mg for I, II, III and IV larval instars, respectively. The mean pupal length and weight of male and female varied significantly when reared at different temperature levels.

Cover Caption

AbstractThe pleasing lacewings (Dilaridae) are a distinctive family of the holometabolous order Neuroptera, mainly characterized by the sexually dimorphic antennae and long tubular ovipositor, which are widely distributed in most zoogeographic regions except for Australia. The female egg‐laying behaviors, and the anatomy of the larval head are reported in detail for understanding their phylogeny and larval adaptation to subterranean habitat (see pages 1445–1463). Photo provided by Yuezheng Tu.

Study of the family Cimbicidae (Insecta: Hymenoptera)

Larvae of Cimbex quadrimaculata (Müller 1766) (Hymenoptera: Cimbicidae) are grayish and black markings. The larval head capsule is black in the first larval instars. It is a pest mainly on almonds but also feeds on apricots, peaches, pears, and cherries in Turkey. There has been no documentation of this species causing severe defoliation in areas where it has been observed. Observations were continued in the orchard until the end. The larvae fed on the edges of the leaves by consuming the whole leaf laminae or leaving only the midrib. The aim of this mini-review was to describe the distribution, life history, and biology of the almond sawfly (Hymenoptera Cimbicidae). Indexed articles, scientific book chapters, theses databases, university dissertations, national and international scientific articles, scientific journals, documents, and academic and scientific journals are available online ResearchGate, HAL, SSRN, Scielo, and Qeios were used. The present work uses the reference of bibliographical research, understood as the act of inquiring and seeking information on a certain subject, through a survey carried out in national and foreign databases, with the objective of detecting what exists of consensus or controversy.

Expression profile and function analysis of MsCSP17 and MsCSP18 in the larval development of Mythimna separata.

Chemosensory proteins (CSPs) were necessary for insect sensory system to perform important processes such as feeding, mating, spawning, and avoiding natural enemies. However, their functions in non-olfactory organs have been poorly studied. To clarify the function of CSPs in the development of Mythimna separata (Walker) larvae, two CSP genes, MsCSP17 and MsCSP18, were identified from larval integument transcriptome dataset. Both of MsCSP17 and MsCSP18 contained four conserved cysteine sites (C × (6)-C × (18)-C × (2)-C), with a signal peptide at the N-terminal. RT-qPCR analysis showed that MsCSP17 and MsCSP18 have different expression patterns among different developmental stages and tissues. MsCSP17 was highly expressed in 1st-4th instar larvae, and MsCSP18 had high expression in adults. Both genes were expressed highly in larval head, thorax, integument and mandible. Moreover, both of MsCSP17 and MsCSP18 were lowly expressed in larval integuments when larvae molted for 6 h and 9 h from 3rd to 4th instar, but highly at the beginning and end phase during molting. After injection of dsMsCSP17 and dsMsCSP18, the expression levels of two genes decreased significantly, with the body weight of larvae decreased, the mortality increased, and the eclosion rate decreased. It was suggested that MsCSP17 and MsCSP18 contributed to the development of M. separata larvae.

Physiological responses of mechanosensory systems in the head of larval zebrafish (Danio rerio).

The lateral line system of zebrafish consists of the anterior lateral line, with neuromasts distributed on the head, and the posterior lateral line, with neuromasts distributed on the trunk. The sensory afferent neurons are contained in the anterior and posterior lateral line ganglia, respectively. So far, the vast majority of physiological and developmental studies have focused on the posterior lateral line. However, studies that focus on the anterior lateral line, especially on its physiology, are very rare. The anterior lateral line involves different neuromast patterning processes, specific distribution of synapses, and a unique role in behavior. Here, we report our observations regarding the development of the lateral line and analyze the physiological responses of the anterior lateral line to mechanical and water jet stimuli. Sensing in the fish head may be crucial to avoid obstacles, catch prey, and orient in water current, especially in the absence of visual cues. Alongside the lateral line, the trigeminal system, with its fine nerve endings innervating the skin, could contribute to perceiving mechanosensory stimulation. Therefore, we compare the physiological responses of the lateral line afferent neurons to responses of trigeminal neurons and responsiveness of auditory neurons. We show that anterior lateral line neurons are tuned to the velocity of mechanosensory ramp stimulation, while trigeminal neurons either only respond to mechanical step stimuli or fast ramp and step stimuli. Auditory neurons did not respond to mechanical or water jet stimuli. These results may prove to be essential in designing underwater robots and artificial lateral lines, with respect to the spectra of stimuli that the different mechanosensory systems in the larval head are tuned to, and underline the importance and functionality of the anterior lateral line system in the larval fish head.

Transcriptome analysis reveals the role of Zelda in the regulation of embryonic and wing development of Tribolium castaneum.

Zinc finger protein (Zelda) of Tribolium castaneum (TcZelda) has been showed to play pivotal roles in embryonic development and metamorphosis. However, the regulatory mechanism of TcZelda associated with these physiology processes is unclear. Herein, the developmental expression profile showed that Zelda of T. castaneum was highly expressed in early eggs. Tissue expression profiling revealed that TcZelda was mainly expressed in the larval head and adult ovary of late adults and late larvae. TcZelda knockdown led to a 95% mortality rate in adults. These results suggested that TcZelda is related to the activation of the zygote genome in early embryonic development. Furthermore, 592 differentially expressed genes were identified from the dsZelda treated group. Compared with the control group, altered disjunction (ALD) and AGAP005368-PA (GAP) in the dsZelda group were significantly down-regulated, while TGF-beta, propeptide (TGF) was significantly up-regulated, suggesting that TcZelda may be involved in insect embryonic development. In addition, the expression of Ubx ultrabithorax (UBX), Cx cephalothorax (CX), En engrailed (EN), and two Endocuticle structural glycoprotein sgabd (ABD) genes were significantly down-regulated, suggesting that they may cooperate with TcZelda to regulate the development of insect wings. Additionally, Elongation (ELO), fatty acid synthase (FAS), and fatty acyl-CoA desaturase (FAD) expression was inhibited in dsZelda insects, which could disturb the lipase signaling pathways, thus, disrupting the insect reproductive system and pheromone synthesis. These results may help reveal the function of TcZelda in insects and the role of certain genes in the gene regulatory network and provide new ideas for the prevention and control of T. castaneum.

Morphometric traits of Dorcus parallelipipedus (Coleoptera: Lucanidae) larvae and adults: can differences between populations be ruled out?

Summary Stag beetles are saproxylic insects that live in deadwood colonised by fungi and other microorganisms. Due to the cryptic habits of these species, little is known on their biological aspects, especially for the immature stage. In this study, we investigated morphometric traits of lesser stag beetle, Dorcus parallelipipedus (Linnaeus, 1758) (Coleoptera: Lucanidae) larvae and adults, with comparisons between populations. Instar identification by measuring larval head capsule was performed, and the relationship between larval body mass and head capsule width was investigated for each surveyed population. To assess the differences among populations, the scaling relationships for adults between their body and mandibles length was also considered, especially for adult males. Morphometric relationships were described by exponential equations. Differences between populations were sometimes limited to one or the other feature, and particularly ascribed to the larval body mass. These differences could be imputed to several factors, among with those of environmental or genetic order. The scaling relationship of lesser stag beetle adult males showed a positive allometry but did not differ among populations, indicating that morphological traits within a species and sex may be biologically retained. Findings are discussed from the perspective of D. parallelipipedus life history.