Distinct communities under the snow: describing characteristics of subnivium arthropod communities.

Arthropods are active during the winter in temperate regions. Many use the seasonal snowpack as a buffer against harsh ambient conditions and are active in a refugium known as the subnivium. While the use of the subnivium by arthropods is well established, far less is known about subnivium community composition, abundance, biomass, and diversity and how these characteristics compare with the community in the summer. Understanding subnivium communities is especially important given the observed and anticipated changes in snowpack depth and duration due to the changing climate. We compared subnivium arthropod communities with those active during the summer using pitfall trapping in northern New Hampshire. We found that compositions of ground-active arthropod communities in the subnivium differed from those in the summer. The subnivium arthropod community featured moderate levels of richness and other measures of diversity that tended to be lower than the summer community. More strikingly, the subnivium community was much lower in overall abundance and biomass. Interestingly, some arthropods were dominant in the subnivium but either rare or absent in summer collections. These putative "subnivium specialists" included the spider Cicurina brevis (Emerton 1890) (Araneae: Hahniidae) and 3 rove beetles (Coleoptera: Staphylinidae): Arpedium cribratum Fauvel, 1878, Lesteva pallipes LeConte, 1863, and Porrhodites inflatus (Hatch, 1957). This study provides a detailed account of the subnivium arthropod community, establishes baseline information on arthropod communities in temperate forests of northeastern North America, and explores the idea of subnivium specialist taxa that are highly active in winter and might be especially vulnerable to climate change.

Appendage abnormalities in spiders induced by an alternating temperature protocol in the context of recent advances in molecular spider embryology.

In the literature there are numerous reports of developmental deformities in arthropods collected in their natural habitat. Since such teratogenically affected individuals are found purely by chance, the causes of their defects are unknown. Numerous potential physical, mechanical, chemical, and biological teratogens have been considered and tested in the laboratory. Thermal shocks, frequently used in teratological research on the spider Eratigena atrica, have led to deformities on both the prosoma and the opisthosoma. In the 2020/2021 breeding season, by applying alternating temperatures (14 °C and 32 °C, changed every 12 h) for the first 10 days of embryonic development, we obtained 212 postembryos (out of 3,007) with the following anomalies: oligomely, heterosymely, bicephaly, schistomely, symely, polymely, complex anomalies, and others. From these we selected six spiders with defects on the prosoma and two with short appendages on the pedicel for further consideration. The latter cases seem particularly interesting because appendages do not normally develop on this body part, viewed as the first segment of the opisthosoma, and appear to represent examples of atavism. In view of the ongoing development of molecular techniques and recent research on developmental mechanisms in spiders, we believe the observed phenotypes may result, at least in part, from the erroneous suppression or expression of segmentation or appendage patterning genes. We consider "knockdown" experiments described in the literature as a means for generating hypotheses about the sources of temperature-induced body abnormalities in E. atrica.

Giant polyploid epidermal cells and male pheromone production in the tephritid fruit fly Eurosta solidaginis (Diptera: Tephritidae).

Eurosta solidaginis males produce large amounts of putative sex pheromone compared to other insect species; however, neither the site of pheromone production nor the release mechanism has been characterized. We compared E. solidaginis males and females, focusing on sexually dimorphic structures that are known to be involved in pheromone production in other tephritid species. Morphological and chemical analyses indicated that the rectum and pleural epidermis are involved in male E. solidaginis pheromone production, storage, or emission. We detected large quantities of pheromone in the enlarged rectum, suggesting that it stores pheromone for subsequent release through the anus. However, pheromone might also discharge through the pleural cuticle with the involvement of unusual pleural attachments of the tergosternal muscles, which, when contracted in males, realign specialized cuticular surface elements and expose less-sclerotized areas of cuticle. In males, pheromone components were also detected in epidermal cells of the pleuron. These cells were 60-100 times larger in mature males than in females and, to our knowledge, are the largest animal epithelial cells ever recorded. Furthermore, because these large cells in males are multinucleated, we presume that they develop through somatic polyploidization by endomitosis. Consequently, the pheromone-associated multinuclear pleural epidermal cells of Eurosta solidaginis may provide an interesting new system for understanding polyploidization.

Temperature fluctuations during embryonic development implicated in a naturally occurring instance of abnormal spinnerets in the spider Australomimetus maculosus (Araneae, Mimetidae).

We record developmental abnormalities of the spinnerets in a field-collected adult male specimen of Australomimetus maculosus. These include (1) a supernumerary right posterior lateral spinneret (PLS), (2) ectopic piriform silk gland spigots and tartipores on the left PLS that are normally restricted to anterior lateral spinnerets (ALSs), and (3) what appear to be ectopic ALS sensilla on the left posterior median spinneret (PMS). Published results of teratological experiments and climate data for the collection site indicate that fluctuating sub- and supra-optimal temperatures during embryogenesis may have been responsible for these anomalies. This specimen thus supports the view that spinneret abnormalities, among other aberrations, may be induced when embryos of entelegyne spiders are exposed to fluctuations between high and low temperatures, whether in the laboratory or, as here, in nature. To our knowledge, the ectopic structures seen on the left PLS and left PMS have not been observed previously. Their locations are consistent with a hypothesis by which only the lateral portion of the araneomorph ALS is serially homologous to the PLS, while the remainder of the ALS, along with the colulus/cribellum, is homologous to the PMS.

Hers and his: Silk glands used in egg sac construction by female spiders potentially repurposed by a 'modern' male spider.

Cylindrical silk gland (CY) spigots distinguish a large clade of modern spiders, the CY spigot clade, which includes all entelegyne spiders and their closest relatives. Following a widespread paradigm, CYs and their spigots are only known to occur in female spiders and they produce silk used in the construction of egg sacs. Here we report the occurrence of a CY spigot or CY nubbin on each posterior median spinneret (PMS) in males (5th stadium and later) of the spider Australomimetus maculosus. Late juvenile males had a CY spigot on each PMS, whereas adult males either had a CY spigot or, more often, a non-functional CY nubbin. This indicates that potential CY use by males is at least largely limited to late juvenile instars and is not involved with egg sac construction. Despite the presence of CY spigots in both sexes, sexual dimorphism with respect to CYs was still evident since males lacked the CY spigot on each posterior lateral spinneret present in late juvenile and adult females, and CY spigots of males never had the wide shaft and opening of adult females. This study adds to our knowledge of spinning apparatus variability in modern spiders and demonstrates an exception to the paradigm that, in the CY spigot clade, such spigots are restricted to female spiders.

Phylogenetic affinities of Phobetinus to other pirate spider genera (Araneae: Mimetidae) as indicated by spinning field morphology.

Spinnerets from Phobetinus sagittifer and an undescribed Phobetinus species were examined by scanning electron microscopy to gain a better understanding of this genus' relationships to other genera in the family Mimetidae. Consistent with placement of Phobetinus in Mimetinae, females possessed two synapomorphies of this subfamily; enlarged cylindrical silk gland spigots with domed shafts and a single cylindrical spigot per posterior lateral spinneret (PLS). Spinning field features overall suggest Phobetinus is most closely related to Mimetus, followed by Australomimetus, then Ero. A possible synapomorphy of a clade including Mimetus and Phobetinus is a pair of modified piriform silk gland spigots on each anterior lateral spinneret of adult males located adjacent to the secondary major ampullate silk gland tartipore. These spigots were present in P. sagittifer; however, similarly positioned spigots in the undescribed species were not obviously modified (i.e., wider or with larger openings relative to the other piriform spigots). Close affinity to Mimetus was also indicated by tartipore-accommodated PLS aciniform silk glands in both Phobetinus species. These have been consistently observed in Mimetus, but not in Australomimetus or Ero. Somatic and genitalic drawings of P. sagittifer are provided to aid identification and similarities are noted between male pedipalps of Mimetus and Phobetinus.

Small organic solutes in sticky droplets from orb webs of the spider Zygiella atrica (Araneae; Araneidae): ß-alaninamide is a novel and abundant component.

In northeastern North America, Zygiella atrica often build their orb webs near the ocean. We analyzed individual field-built Z. atrica webs to determine if organic low-molecular-mass solutes (LMM) in their sticky droplets showed any unusual features not previously seen in orb webs of other species living in less salty environments. While two of the three most abundant organic LMM (putrescine (butane-1,4-diamine) and GABamide (4-aminobutanamide)) are already well-known from webs of inland spiders, the third major LMM, ß-alaninamide (3-aminopropanamide), a homolog of GABamide, has not been detected in sticky droplets from any other araneoid spiders (27 species). It remains to be established, however, whether or not use of ß-alaninamide is related to proximity to saltwater. We observed variability in organic LMM composition in Z. atrica webs that appeared to be influenced more by an undetermined factor associated with different collecting locations and/or collection dates than by different genders or instars. Shifts in composition when adult females were transferred from the field to the laboratory were also observed. Structural similarities and inverse correlations among ß-alaninamide, GABamide, and N-acetylputrescine suggest that they may form a series of LMM fulfilling essentially the same, as yet unknown, role in the webs of those species in which they occur.

Free amino acids in spider hemolymph.

We examined the free amino acid composition of hemolymph from representatives of five spider families with an interest in knowing if the amino acid profile in the hemolymph of orb-web-building spiders reflects the high demands for small organic compounds in the sticky droplets of their webs. In nearly all analyses, on both orb and non-orb builders, glutamine was the most abundant free amino acid. Glycine, taurine, proline, histidine, and alanine also tended to be well-represented in orb and non-orb builders. While indications of taxon-specific differences in amino acid composition were observed, it was not apparent that two presumptive precursors (glutamine, taurine) of orb web sticky droplet compounds were uniquely enriched in araneids (orb builders). However, total amino acid concentrations were invariably highest in the araneids and especially so in overwintering juveniles, even as several of the essential amino acids declined during this winter diapause. Comparing the data from this study with those from earlier studies revealed a number of discrepancies. The possible origins of these differences are discussed.

Changes in composition of spider orb web sticky droplets with starvation and web removal, and synthesis of sticky droplet compounds.

The sticky spiral of araneoid spider orb webs consists of silk fibers coated with adhesive droplets. The droplets contain a variety of low-molecular-mass compounds (LMM). Within a species, a fairly consistent ratio of LMM is often observed, but substantial variability can exist. To gain insight into factors influencing LMM composition, spiders of three araneid species were starved and LMM from their webs were analyzed for changes in composition. To determine if these changes were consistent with the spider's ability to synthesize the different organic LMM, synthetic capacities were estimated following the feeding of radiolabeled metabolites. Some changes in droplet composition were broadly consistent with differing synthetic capacities: molar percentages of less readily synthesized compounds (e.g. choline, isethionate, N-acetyltaurine) typically declined with starvation, at least during a portion of the imposed fast, while more readily synthesized compounds (e.g. GABamide, glycine) tended to increase. Most striking was the apparent partial substitution of N-acetylputrescine by the more readily synthesized GABamide in fasting Argiope trifasciata. However, departures from expected compositional shifts demonstrated that synthetic capacity alone does not adequately predict sticky droplet compositional shifts with starvation. Moreover, feeding controls exhibited some changes in composition similar to starving spiders. As the webs of both feeding and starving spiders were removed for chemical analysis and could not be recycled, the loss of LMM contained in these webs likely contributed to similarities between treatments. In addition, feeding spiders molted, oviposited and/or built heavier webs. The added metabolic demands of these activities may have contributed to changes in composition similar to those resulting from starvation.

Astacin family metallopeptidases and serine peptidase inhibitors in spider digestive fluid.

Digestive fluid of the araneid spider Argiope aurantia is known to contain zinc metallopeptidases. Using anion-exchange chromatography, size-exclusion chromatography, sucrose density gradient centrifugation, and gel electrophoresis, we isolated two lower-molecular-mass peptidases, designated p16 and p18. The N-terminal amino acid sequences of p16 (37 residues) and p18 (20 residues) are 85% identical over the first 20 residues and are most similar to the N-terminal sequences of the fully active form of meprin (beta subunits) from several vertebrates (47-52% and 50-60% identical, respectively). Meprin is a peptidase in the astacin (M12A) subfamily of the astacin (M12) family. Additionally, a 66-residue internal sequence obtained from p16 aligns with the conserved astacin subfamily domain. Thus, at least some spider digestive peptidases appear related to astacin of decapod crustaceans. However, important differences between spider and crustacean metallopeptidases with regard to isoelectric point and their susceptibility to hemolymph-borne inhibitors are demonstrated. Anomalous behavior of the lower-molecular-mass Argiope peptidases during certain fractionation procedures indicates that these peptidases may take part in reversible associations with each other or with other proteins. A. aurantia digestive fluid also contains inhibitory activity effective against insect digestive peptidases. Here we present evidence for at least thirteen, heat-stable serine peptidase inhibitors ranging in molecular mass from about 15 to 32 kDa.

Selected aspects of spinning apparatus development in Araneus cavaticus (Araneae, Araneidae).

In the first half of this century, several workers observed small, seemingly glandular structures attached to the ampullate glands of spiders. Hence, they were termed accessory ampullate glands. In juvenile Araneus cavaticus, two pairs of these structures are present (starting at least with third instars), one pair attached to the major ampullate (MaA) glands and the other pair attached to the minor ampullate (MiA) glands. In adults, two pairs of accessory MaA glands and two pairs of accessory MiA glands are present. The two latter-formed pairs of accessory ampullate glands are clearly the remnants of those ampullate glands which atrophy shortly after adulthood is reached. Morphological similarities between these accessory ampullate glands and those present in juveniles provide an indication that the latter also have their origin in functional ampullate glands. A reduction in the number of ampullate glands following the last molt occurs in many spiders. The reason(s) for these reductions is unknown. In penultimate spiders close to ecdysis, we have observed that while the larger pairs of MaA and MiA glands (those that are retained in the adult) are undergoing molt-related changes which apparently render them nonfunctional, their smaller counterparts are seemingly unaffected and functional. This raises the possibility that the principal role of the smaller ampullate glands may be to assume functions during the pre-ecdysial period which are normally in the domain of the larger ampullate glands. If true, then their degeneration after the last molt would make economic sense. The presence of cylindrical spigots in juvenile females starting with fourth instars is documented.