Comb cells and puparial silk in the oriental hornet nest: Structure and function
Authors: Ishay Jacob S. , Ganor Eliezer 
Published: February 2005

AbstractAdult hornets (Vespa orientalis; Hymenoptera, Vespinae) build the brood combs out of organic or mineral matter. The cement that serves to glue the building material together is secreted in their saliva, the latter hardening within seconds to form fibers or plates. This saliva‐derived spittle overlies and unites the building particles laminally and vertically. The hornet larvae spin a cocoon within the brood cells, which is largely fastened to and supported by the cell walls and is composed of a network of silk fibers and interlinking flat surfaces. On the outside of the cocoon fibers are spherical button‐like structures that are very rich in phosphorus. The chemical composition of the adult salivary cement and the larval cocoon fibers is similar: both contain the elements P, Mg, S, Cl, K, and Ca. The possible biological significance of these findings is discussed.Among social insects belonging to the Hymenoptera, there are three main groups that build a multitude of cells, namely, the paper wasps (Polistinae), the other social wasps or hornets (Vespinae), and the social bees (Apinae). The constructed cells serve mainly for rearing the brood, but in many instances (particularly among various Apinae) they also serve for storing honey and pollen. For building material, Apinae rely primarily on beeswax (a product secreted by the bee itself to which various amounts of plant resins are added). On the other hand, wasps (Polistinae and Vespinae) build mainly from matter collected in the nearby environment, be it organic matter such as tree bark, mineral matter, or a combination of the two.Much information has accumulated on comb and cell building among these insect groups (e.g., Wheeler, '23; Van der Vecht, '57, '65; Lindauer, '61; Michener, '61; Kemper and Döhring, '67; Wilson, '71; Guiglia, '72; Spradbery, '73; Edwards, '80; Brian, '83; Schremmer et al., '85). Species of Polistinae and Vespinae are prevalent in forest areas in Southeast Asia and in central and South America or in the temperate regions in both the northern and southern hemispheres, and they mainly use vegetable matter to build their combs. In contrast, species prevalent in the Mediterranean region, which is dry and relatively unvegetated during the wasp and hornet active season, rely more on nearby mineral than on plant matter. Comb building in the Oriental hornet is well known (Darchen, '64; Ishay et al., '67; Schaudinischky and Ishay, '68; Ishay, '73, '75a, b, '76; Ishay and Sadeh, '75, '76; Ishay and Perna, '79; Ishay et al., '82). Recently Ganor et al. ('86) described the cell wall in the V. orientalis comb, showing it to be composed of mineral particles collected near the subterranean nest, in contrast to the comb of two European species (V. crabro and Vespula) (Paravespula) germanica, which is built primarily of organic matter. Regardless of whether the building material is mineral or organic, the hornets utilize particles of sand or other minerals or pieces of wood wrapped and melded together by saliva. However, nothing is known about the cement or mortar used to glue together these “bricks.” The present investigation was initiated to increase our knowledge of this cementing substance.Combs of V. orientalis were collected from fields in the Tel‐Aviv district in 1987 during the active season, which extends over most of the summer months. Because the combs are easily damaged, care was taken to remove them intact from the natural nest. Once removed, the combs were cleared of the existing brood (eggs, larvae, or pupae) and then stored carefully in dry glass vessels until examined. For the present study, combs were collected only from nests in Khamra soil, which is common in the Tel‐Aviv district and along the coastal area of Israel. To investigate the morphology of the cement material in the comb, strips of comb wall as well as segments of the pupal silk dome were removed from each comb and prepared for examination. The comb wall strips were cut to a size of 3 × 6 mm and fastened to the stub of a scanning electron microscope (SEM). The silk pieces were processed in two ways: (1)strips of 3 × 4 mm were fastened onto a stub with the convex (outer surface) facing up or (2) with the concave (interior) side upward.Silk strips were boiled for 2 hours in distilled water to remove all impurities, such as foreign matter adhering to the silk secreted by the larvae. Examination of these silk fibres was carried out in the three ways: (1)SEM micrography of the exterior (white portion) of the silk dome; (2) micrography of the interior; (3) micrography of both exterior and interior aspects after boiling for 2 hours in distilled water to remove water‐suspended foreign materials that may have clung to the silk dome in the course of ordinary nest activity.The chemical composition of selected specimens was investigated by x‐ray analysis. They were done on JEOL 840 Scanning Electron Microscope (SEM) equipped with Link 10,000 Energy‐Dispersive System (EDS). (With the EDS System, the spectrometer separates the elements according to energy rather than wavelength). Quantitative analysis was by ZAF4 program. Five strips of comb cell wall were examined from a randomly selected comb, each comb from a different nest. Micrographs were taken of isolated silk samples from each of the examined combs.