Age dependent changes in histamine content of venom of queen and worker honey bees

We have compared the variation in histamine content of the venom of worker honey bees maintained in the field with those in an indoor flight room. We find that the temporal changes in venom histamine content are parallel under the different conditions.We have measured the histamine content of different regions of the venom system of virgin queen bees and found temporal changes in histamine concentration which parallel those seen in workers. While the absolute amounts of histamine in the queen venom system exceed those measured in workers, we estimate that the histamine concentration in queen and worker venom is similar.The LD₅₀ of histamine, injected into the haemocoele of worker bees in saline, is found to be 3.2 μg/worker.     

Cover Caption

The Western honey bee, Apis mellifera (L.), is perhaps the most beneficial insect we know, mainly because of the pollination services it provides to fruits and vegetables. Honey bee workers show changes in behaviors as they age. Young bees typically are “nurses” and perform in‐hive tasks such as feeding larvae and take care of the queen, old bees become foragers and bring in food sources (nectar, pollen, and water) or propolis. Methoprene has been known to accelerate worker development so bees become foragers earlier, but its mechanisms were not known. In this study Huang et al. show that most likely methoprene works directly on hormone receptors to mimic juvenile hormone, in causing bees to forage early (pages 235–240). Photo by Zachary Y. Huang. [Correction added on 17 April 2018, after first online publication: Cover caption has been revised.]     

Effect of age,behaviour and social environment on honey bee brain plasticity

We examined the effects of behaviour, age and social environment on mushroom body volume in adult bees. The mushroom bodies are regions of the central brain important for sensory integration and learning. Their volume was influenced by behaviour throughout life: always larger in forager bees than age-matched nurse bees, even in old bees up to 93 days of age as adults. Mushroom body development was influenced by the social environment in the first 8 days of adult life, with different environments having markedly different effects on mushroom body size. Compared to hive-reared bees, isolation slowed mushroom body growth, but bees reared in isolation confined with a single dead bee showed a dramatic increase in mushroom body volume comparable to that seen in active foragers. Despite their precocious mushroom body development, these bees did not show improved performance in an olfactory learning test. Since simple environmental manipulations can both accelerate and delay mushroom body growth in young bees, and since mushroom body volume is sensitive to behaviour throughout life, the honey bee has great potential as a model for exploring the interactions between environment, behaviour and brain structure.     

The evolution of floral sonication,a pollen foraging behavior used by bees (Anthophila)

Over 22,000 species of biotically pollinated flowering plants, including some major agricultural crops, depend primarily on bees capable of floral sonication for pollination services. The ability to sonicate (“buzz”) flowers is widespread in bees but not ubiquitous. Despite the prevalence of this pollinator behavior and its importance to natural and agricultural systems, the evolutionary history of floral sonication in bees has not been previously studied. Here, we reconstruct the evolutionary history of floral sonication in bees by generating a time‐calibrated phylogeny and reconstructing ancestral states for this pollen extraction behavior. We also test the hypothesis that the ability to sonicate flowers and thereby efficiently access pollen from a diverse assemblage of plant species, led to increased diversification among sonicating bee taxa. We find that floral sonication evolved on average 45 times within bees, possibly first during the Early Cretaceous (100–145 million years ago) in the common ancestor of bees. We find that sonicating lineages are significantly more species rich than nonsonicating sister lineages when comparing sister clades, but a probabilistic structured rate permutation on phylogenies approach failed to support the hypothesis that floral sonication is a key driver of bee diversification. This study provides the evolutionary framework needed to further study how floral sonication by bees may have facilitated the spread and common evolution of angiosperm species with poricidal floral morphology.     

Phylogenetic analysis of the corbiculate Apinae based on morphology of the sting apparatus (Hymenoptera: Apidae)

This study aimed to test the various competing hypotheses regarding the relationships among the four tribes of corbiculate apine bees (Euglossini “orchid bees”, Bombini “bumble bees”, Meliponini “stingless bees”, and Apini “honey bees”) with a completely new set of previously unstudied morphological characters derived from the sting apparatus. The result was one most parsimonious tree of 49 steps, CI = 89, RI = 93 that is perfectly congruent with most studies based on morphological and combined morphological/molecular data, i.e., Euglossini + (Bombini + (Meliponini + Apini)), supporting a well accepted scenario of social evolution for these bees. This data matrix was then combined with other published matrices for this group in order to perform simultaneous analyses. The problem of how to best combine the multiple matrices that did not use the same exemplars was investigated. © The Willi Hennig Society 2007.     

The effects of honey bee (Apis mellifera L.) queen reproductive potential on colony growth

Reproduction in species of eusocial insects is monopolized by one or a few individuals, while the remaining colony tasks are performed by the worker caste. This reproductive division of labor is exemplified by honey bees (Apis mellifera L.), in which a single, polyandrous queen is the sole colony member that lays fertilized eggs. Previous work has revealed that the developmental fate of honey bee queens is highly plastic, with queens raised from younger worker larvae exhibiting higher measures in several aspects of reproductive potential compared to queens raised from older worker larvae. Here, we investigated the effects of queen reproductive potential (“quality”) on the growth and winter survival of newly established honey bee colonies. We did so by comparing the growth of colonies headed by “high-quality” queens (i.e., those raised from young worker larvae, which are more queen-like morphologically) to those headed by “low-quality” queens (i.e., those raised from older worker larvae, which are more worker-like morphologically). We confirmed that queens reared from young worker larvae were significantly larger in size than queens reared from old worker larvae. We also found a significant positive effect of queen grafting age on a colony’s production of worker comb, drone comb, and stored food (honey and pollen), although we did not find a statistically significant difference in the production of worker and drone brood, worker population, and colony weight. Our results provide evidence that in honey bees, queen developmental plasticity influences several important measures of colony fitness. Thus, the present study supports the idea that a honey bee colony can be viewed (at least in part) as the expanded phenotype of its queen, and thus selection acting predominantly at the colony level can be congruent with that at the individual level.     

Morphological,ultrastructural, histochemical and electrophoretic studies on the venom system of Nasonia vitripennis walker (hymenoptera: Pteromalidae)

The venom system of Nasonia vitripennis is well-developed and composed of an unbranched acid gland and associated reservoir. Fine-structural, histochemical and electrophoretic studies indicate that the venom is produced by two protein-secreting epithelia. The bulk of the venom is synthesised in the columnar cells of the acid gland and discharged via “vesicular organelles” and the efferent ductular system into the lumen of the reservoir. The acid gland also contains squamous chitogenous cells, situated either around the central lumen or interposed between the bases of the columnar cells. Once within the reservoir, the venom is probably activated by enzymatic secretions from the reservoir secretory cells. Each of these cells has a “vesicular organelle” but, in contrast to the columnar cells of the acid gland, the cytoplasm contains a preponderance of free ribosomes, and protein segregation apparently occurs outside the Golgi complexes. The venom is expelled through the efferent discharge duct by muscular contractions, which open the duct lumen and bring it into contact with the funnel of the ovipositor. Excessive distortion of the duct is prevented by a massive ventral ligament.     

Neuromuscular and hormonal control of post-eclosion processes in flies

Flies (exemplified by Sarcophaga bullata) expand after eclosion from the puparium by processes of “pulsing” (slow rhythmical abdominal contractions) and “pumping” of air (fast rhythmical contractions of the cibarial pump). Pulsing and pumping are inhibited if a newly eclosed fly is kept in an enclosed space (sand, a glass tube, an empty puparium). This inhibition no longer applies if such flies are injected with either hemolymph from flies 10–15 min old or cAMP, or are confined at the age of 10 min. This suggests a hormonal control of pulsing and pumping. Pumping alone, without pulsing, occurs in flies treated with certain paralyzing agents like ether, tetrodotoxin, bee venom, or “FlyNap,” or have the connectives in the neck cut or interrupted by cauterization. Application of FlyNap or neck cauterization leads to excessive pumping which results in bloating. Expansion by bloating is confined to the soft membranes, leaving sclerites and wings largely unexpanded. The function of pulsing is probably that of “plasticizing” and stretching the cuticle to make it respond to increased steady pressure by air-pumping. Flies ligated at the proboscis show almost regular pulsing and pumping, but without intake of air, and consequently no expansion. Cuticle (sclerites) and wings, however, become plasticized. Some plasticization occurs even in the absence, or reduction, of pulsing (in a neck-cauterized fly), brought about by a hormonal process. Eclosion from the puparium is also initiated by a hormonal action. Thus, the following processes during fly emergence are controlled by hormones: eclosion proper, pulsing, pumping, plasticization, and tanning. These hormones are separate entities, with the possible exception of the pulsing/pumping hormone(s).     

Cellular energy metabolism maintains young status in old queen honey bees (Apis mellifera)

Trophocytes and oenocytes of queen honey bees are used in studies of cellular longevity, but their cellular energy metabolism with age is poorly understood. In this study, the molecules involved in cellular energy metabolism were evaluated in the trophocytes and oenocytes of young and old queen bees. The findings indicated that there were no significant differences between young and old queen bees in β‐oxidation, glycolysis, and protein synthesis. These results indicate that the cellular energy metabolism of trophocytes and oenocytes in old queen bees is similar to young queen bees and suggests that maintaining cellular energy metabolism in a young status may be associated with the longevity of queen bees. Fat and glycogen accumulation increased with age indicating that old queen bees are older than young queen bees.     

Invasive ant (<Emphasis Type="Italic">Anoplolepis gracilipes</Emphasis>) disrupts pollination in pumpkin

Yellow crazy ant (Anoplolepis gracilipes (F. Smith); “YCA”) is known for its aggressive predatory ability and ability to exert exploitation competition on both native and other invasive ants via floral nectar. We argue that YCA invasion can exert both interference and exploitation competition on legitimate pollinators. In pumpkin fields (Cucurbita maxima L.) of south India, YCA infested the flowers, particularly the pistillate flowers, for nectar foraging. Pumpkin is a honey bee-mediated cross-pollinated monoecious plant that produces disproportionately very few pistillate flowers. We hypothesize that YCA presence in the flowers can affect the visitation rate and foraging time of honey bees in the flowers, the fruit set in pumpkins, and can exert predatory pressure on the honey bees if the bees linger in ant-colonized flowers. Both YCA and honey bees preferred to forage on the limited pistillate flowers in the plants. After colonizing the flowers, YCA did not retreat for hours, even upon disturbance by competitors, such as honey bees. Both the visitation frequency and the foraging time of honey bees were drastically reduced in ant-colonized flowers, and none of the ant-colonized flowers developed into fruits, suggesting that the YCA exert both an ecological and evolutionary pressure on pumpkin. The ants preyed upon about 17% of the honey bees that lingered in ant-colonized flowers, and the time the bees spent foraging predicted the fate of the bees. Exploitation competition exerted by the YCA on pumpkin may have far-reaching consequences for the pollination and productivity of this cash crop.     

Differentiation rather than aging of muscle stem cells abolishes their telomerase activity

A general feature of stem cells is the ability to routinely proliferate to build, maintain, and repair organ systems. Accordingly, embryonic and germline, as well as some adult stem cells, produce the telomerase enzyme at various levels of expression. Our results show that, while muscle is a largely postmitotic tissue, the muscle stem cells (satellite cells) that maintain this biological system throughout adult life do indeed display robust telomerase activity. Conversely, primary myoblasts (the immediate progeny of satellite cells) quickly and dramatically downregulate telomerase activity. This work thus suggests that satellite cells, and early transient myoblasts, may be more promising therapeutic candidates for regenerative medicine than traditionally utilized myoblast cultures. Muscle atrophy accompanies human aging, and satellite cells endogenous to aged muscle can be triggered to regenerate old tissue by exogenous molecular cues. Therefore, we also examined whether these aged muscle stem cells would produce tissue that is “young” with respect to telomere maintenance. Interestingly, this work shows that the telomerase activity in muscle stem cells is largely retained into old age wintin inbred “long” telomere mice and in wild‐derived short telomere mouse strains, and that age‐specific telomere shortening is undetectable in the old differentiated muscle fibers of either strain. Summarily, this work establishes that young and old muscle stem cells, but not necessarily their progeny, myoblasts, are likely to produce tissue with normal telomere maintenance when used in molecular and regenerative medicine approaches for tissue repair. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

FLORAL PAEDOMORPHY LEADS TO SECONDARY SPECIALIZATION IN POLLINATION OF MADAGASCAR DALECHAMPIA (EUPHORBIACEAE)

The traditional evolutionary interpretation of Von Baer's “laws” of embryology is that retention of early developmental forms into adulthood (paedomorphosis) leads to the evolution of simpler or more generalized morphology and ecology. Here we show that paedomorphosis can also be involved in an increase in ecological specialization, in this case of plant–pollinator relationships. A paedomorphic transition from generalized pollination (by several functional types of pollinators) to specialized pollination (by one or a few species in one functional type) occurred in a clade of endemic Madagascar vines (Dalechampia spp., Euphorbiaceae). This evolutionary transition involved staminate flowers that fail to develop “normally,” instead holding mature pollen inside virtually unopened, bud‐like flowers. This paedomorphic morphology restricts reward access to “buzz‐pollinating” bees, including Xylocopa species (carpenter bees), which can remove pollen by sonication. This is one of very few reports of paedomorphic specialization, and, as far as we are aware, the first documented case of a rapid reversal to specialized pollination in a lineage of plants that had previously switched from specialized to generalized pollination in conjunction with dispersing to a new region.     

Studies on ribonucleic acid synthesis in the venom glands of Vipera palaestinae (Ophidia, Reptilia)

RNA metabolism in the venom glands of Vipera palaestinae was studied at different stages after manual extraction of the venom (milking). The rate of (32)P incorporation into gland RNA was found to be maximal at 1-4 days after milking in correlation with the height of the secretory epithelium. Venom production attained a maximum only after 8-16 days, in parallel with the accumulation of stable species of cellular RNA.     

In situ extraction (milking) of the two promising Botryococcus braunii strains Showa and Bot22 under optimized extraction time

Journal of Applied Phycology - In situ extraction or “milking” of microalgae is a promising approach to reduce downstream costs in order to produce low-value substances such as lipids...     

Changes in weight of the pharyngeal gland and haemolymph titres of juvenile hormone,protein and vitellogenin in worker honey bees

Four physiological parameters (haemolymph-juvenile hormone titre, protein concentration, vitellogenin concentration, and pharyngeal gland dry weight) were examined in the following categories of queenright adult worker bees: summer bees 1–40 days old, winter bees 80–130 days old, 12–100-day old bees at the beginning of winter, 100–195-day old bees at the end of winter, and 1–100-day old bees experimentally induced to live longer in summer.In contrast to the continuously increasing titre of juvenile hormone in ageing summer bees, winter bees kept a constant low level. In bees at the beginning of winter, the hormone titre never reached high values. However, at the end of winter it rose from a low to a high level, comparable with the high titre of 24–40-day old summer bees. In experimentally induced longlived bees in summer, the juvenile hormone titre did not increase as in normal summer bees but remained low as in bees at the beginning of winter. Among the known natural juvenile hormones, only juvenile hormone III was present in the haemolymph of winter bees.The results support the hypothesis of polyphenism being regulated by the titre of juvenile hormone in the haemolymph.     

Laboratory evolution of longevity in the bean weevil (Acanthoscelides obtectus)

In this study we examined the effects of long-term selection on early and late reproduction in the bean weevil. The pure lines and the hybrids between the lines within a selection regime were compared for longevity, early and late female fecundity, male mating ability, pre-adult developmental time and wet adult weight. Comparison of hybrid with pure lines provided some evidence for inbreeding despression in the lines from both selection regimes. We found that virgin and mated adults of both sexes from the “old” lines lived longer than “young” line beetles. Comparisons of the hybrid “young” with hybrid “old” lines revealed a trade-off between early and late fecundity of females. For noncompetitive mating ability of males there was no difference between the lines with different rates of senescence when the males were young. But, when the males were older, beetles from the lines selected for delayed senescence expressed superior mating ability. In addition, the “old” line beetles take longer to develop and are heavier than those from “young” line beetles. Although these data suggest that shorter pre-adult developmental time may imply more rapid senescence, there is the possibility of inadvertent selection for rapid development in the “young” lines and this complicates the interpretation of the observed trade-off between the pre-adult and adult performances.     

Characterization of ribonucleic acids from the venom glands of Crotalus durissus terrificus (Ophidia, Reptilia) after manual extraction of the venom. Studies on template activity and base composition

RNA synthesis in the venom glands of Crotalus durissus terrificus was stimulated by the manual extraction of the venom (milking). RNA was extracted from venom glands activated by milking and fractionated by centrifugation through sucrose density gradients. Template activity for protein synthesis and base composition of the RNA fractions were studied. RNA fractions that sediment between 18S and 4S had the highest template activity. The base composition analysis indicated that the 28S and 18S rRNA have a C+G content of 65.4 and 58% respectively. The ;melting' temperature (T(m)) of DNA in 0.15m-NaCl-0.015m-trisodium citrate, pH7.0, was 85 degrees C, corresponding to a C+G content of 38%. The base ratio of the RNA fractions that showed a high template activity was intermediate between that of rRNA and homologous DNA. The possible role of these fractions in the synthesis of the two main toxins (crotoxin and crotamine) of the South American rattlesnake's venom is discussed.     

Social Modulation of Stress Reactivity and Learning in Young Worker Honey Bees

Alarm pheromone and its major component isopentylacetate induce stress-like responses in forager honey bees, impairing their ability to associate odors with a food reward. We investigated whether isopentylacetate exposure decreases appetitive learning also in young worker bees. While isopentylacetate-induced learning deficits were observed in guards and foragers collected from a queen-right colony, learning impairments resulting from exposure to this pheromone could not be detected in bees cleaning cells. As cell cleaners are generally among the youngest workers in the colony, effects of isopentylacetate on learning behavior were examined further using bees of known age. Adult workers were maintained under laboratory conditions from the time of adult emergence. Fifty percent of the bees were exposed to queen mandibular pheromone during this period, whereas control bees were not exposed to this pheromone. Isopentylacetate-induced learning impairments were apparent in young (less than one week old) controls, but not in bees of the same age exposed to queen mandibular pheromone. This study reveals young worker bees can exhibit a stress-like response to alarm pheromone, but isopentylacetate-induced learning impairments in young bees are suppressed by queen mandibular pheromone. While isopentylacetate exposure reduced responses during associative learning (acquisition), it did not affect one-hour memory retrieval.     

Trap-nesting bees and wasps colonizing set-aside fields: succession and body size,management by cutting and sowing

Trap-nesting bees and wasps (Hymenoptera Aculeata) colonizing crop and fallow fields in an agricultural landscape were studied using 20 sown fields (pea, barley, rye, clover-grass mixtures, Phacelia tanacetifolia) and 20 fields with naturally developed vegetation (1- and 2-year old fields, both mown and unmown, and old meadows). Fourteen species of Apoidea, 4 of Sphecidae, 1 of Eumenidae and 4 of parasitoids were reared from reed nests exposed in these 40 fields of 10 field-types. Fields with naturally developed vegetation had twice as many species as sown fields, due to the distribution pattern of the 14 bee species, whereas the 9 predatory species (wasps and parasitoids) showed a rather uniform distribution. None of the trap-nesting bees were found in Phacelia fields, despite contrasting expectations of beekeepers. Old meadows showed a particularly high abundance and species richness, since only 10% of all traps were exposed, but 32% of all bee nests were sampled in old meadows, including 4 bee species that were not found elsewhere. Accordingly, species richness of fields with naturally developed vegetation showed a significant increase with age. Variability in Hymenoptera species numbers could be explained by corresponding differences in plant species numbers. The alternative hypothesis that field size or field connectivity influenced species richness was not supported. Habitats with great floral diversity appeared to offer better and richer food resources for the flower-visiting bees, whereas food availability apparently did not influence predatory wasps. The bees Osmia caerulescens and Megachile versicolor that had colonized early-successional fields took twice as long to provision cells as those that colonized late-successional meadows characterized by a greater plant species richness. In contrast, the eumenid wasp Ancistrocerus gazella took a similar period of time to provision cells in both field types. In addition, bee and wasp species of plant-species-poor fields were on average significantly larger than those of plant-species-rich fields. Thus, body size appeared to be a good predictor of colonization ability. Management by cutting greatly increased plant species richness in early-successional set-aside fields and thus doubled species richness of bees. Cutting of early-successional habitats can be expected to benefit insects and plants in general, whereas older grassland should show the greatest insect diversity when both mown and unmown parts are present.