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We investigated the effect of adult honey bee pollen nutrition on the flight performance of honey bees. Therefore, caged bees were allowed to perform 30 min of defecation/training flights every second day before flight performance of pollen-fed bees and pollen-deprived bees older than 16 days were compared in a flight mill. We first fed 10 µL of 1 M glucose solution to bees, and after they metabolized this during flight, they were fed 10 µL of 2 M glucose solution for a second flight test. Pollen-deprived bees flew longer and further than pollen-fed bees in both flights. Pollen-fed bees flew faster in the early period at the beginning of flights, whereas pollen-deprived bees were faster in the final phases. Pollen-fed bees were able to raise their maximum flight speed in 2 M glucose solution flights, whereas pollen-constraint bees were not. The two groups did not differ in abdomen fresh weight, but the fresh weight of the head and thorax and dry weight of the head, thorax and abdomen were higher in pollen-fed bees. In a second experiment, we constrained pollen consumption of caged bees during the first 7 days and compared daily consumption of bees from day 8-16 to consumption of bees unrestricted in pollen. We found that pollen-deprived bees perceive the pollen shortage and try to compensate for their needs by consuming significantly more pollen at the later phase of their life than pollen-fed bees of the same age. Still, bees constrained from pollen in the first 7 days did only reach 51.1% of the lifetime consumption of unconstrained bees. This shows that bees can sense the need for essential nutrients from pollen, but their physiological apparatus does not allow them to fully compensate for their early life constraint. Pollen deprivation only in the first 7 days of worker life likewise significantly reduced fresh and dry weights of the body sections (head, thorax, and abdomen) and survival. This underlines the importance of protein consumption in a short critical period early in adult bees' lives for their development and their performance later in life., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Brodschneider, Omar and Crailsheim.)

A diverse supply of pollen is an important factor for honey bee health, but information about the pollen diversity available to colonies at the landscape scale is largely missing. In this COLOSS study, beekeeper citizen scientists sampled and analyzed the diversity of pollen collected by honey bee colonies. As a simple measure of diversity, beekeepers determined the number of colors found in pollen samples that were collected in a coordinated and standardized way. Altogether, 750 beekeepers from 28 different regions from 24 countries participated in the two-year study and collected and analyzed almost 18,000 pollen samples. Pollen samples contained approximately six different colors in total throughout the sampling period, of which four colors were abundant. We ran generalized linear mixed models to test for possible effects of diverse factors such as collection, i.e., whether a minimum amount of pollen was collected or not, and habitat type on the number of colors found in pollen samples. To identify habitat effects on pollen diversity, beekeepers' descriptions of the surrounding landscape and CORINE land cover classes were investigated in two different models, which both showed that both the total number and the rare number of colors in pollen samples were positively affected by 'urban' habitats or 'artificial surfaces', respectively. This citizen science study underlines the importance of the habitat for pollen diversity for bees and suggests higher diversity in urban areas.

The honeybee nest parasite Aethina tumida (small hive beetle) uses behavioural mimicry to induce trophallactic feeding from its honeybee hosts. Small hive beetles are able to induce honeybee workers to share the carbohydrate-rich contents of their crops, but it is not clear whether the beetles are able to induce to workers to feed them the protein-rich hypopharyngeal glandular secretions fed to the queen, larvae and other nest mates. Protein is a limiting macronutrient in an insect's diet, essential for survival, growth and fecundity. Honeybees obtain protein from pollen, which is consumed and digested by nurse bees. They then distribute the protein to the rest of the colony in the form of hypopharyngeal gland secretions. Using 14 C-phenylalanine as a qualitative marker for protein transfer, we show that small hive beetles successfully induce worker bees to feed them the protein-rich secretions of their hypopharyngeal glands during trophallaxis, and that females are more successful than males in inducing the transfer of these protein-rich secretions. Furthermore, behavioural observations demonstrated that female beetles do not preferentially interact with a specific age cohort of bees when soliciting food, but males tend to be more discriminant and avoid the more aggressive and active older bees., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2021. Published by The Company of Biologists Ltd.)

Austrian beekeepers participated in the "C.S.I. Pollen" study as citizen scientists and collected pollen from honey bee colonies in hive mounted traps every three weeks from April to September in 2014 and 2015 to uncover the seasonal availability of pollen sources for bees. 1622 pollen samples were collected and analysed using palynological light microscopy to the lowest taxonomic level possible. For 2014 and 2015 combined, 239 pollen types from more than 85 families were detected. 'Various unknown' species, Taraxacum-form and Plantago spp. were the pollen types collected by the majority of colonies (occurrence), whereas the most pollen grains collected were from Trifolium repens-form, Plantago spp. and Salix spp. (abundance). In spring, trees were found to be the most abundant pollen source, whereas in summer herbs dominated. On average, a colony collected pollen from 16.8 ± 4.7 (2014) and 15.0 ± 4.4 (2015) pollen types per sampling. Those numbers, however, vary between sampling dates and indicate a seasonal pattern. This is also supported by Simpson's diversity index, which was on median 0.668. In both years, 50.0% of analysed pollen samples were partially (>50%) and 4.2% were highly monofloral (i.e. containing >90% of one pollen type). Prevalence of monofloral pollen samples peaked at the beginning and the end of the season, when pollen diversity was the lowest.

Austrian beekeepers frequently suffered severe colony losses during the last decade similar to trends all over Europe. This first surveillance study aimed to describe the health status of Austrian bee colonies and to analyze the reasons for losses for both the summer and winter season in Austria. In this study 189 apiaries all over Austria were selected using a stratified random sampling approach and inspected three times between July 2015 and spring 2016 by trained bee inspectors. The inspectors made interviews with the beekeepers about their beekeeping practice and the history of the involved colonies. They inspected a total of 1596 colonies for symptoms of nine bee pests and diseases (four of them notifiable diseases) and took bee samples for varroa mite infestation analysis. The most frequently detected diseases were three brood diseases: Varroosis, Chalkbrood and Sacbrood. The notifiable bee pests Aethina tumida and Tropilaelaps spp. were not detected. During the study period 10.8% of the 1596 observed colonies died. Winter proved to be the most critical season, in which 75% of the reported colony losses happened. Risks for suffering summer losses increased significantly, when colonies were weak in July, had queen problems or a high varroa mite infestation level on bees in July. Risks for suffering winter losses increased significantly, when the colonies had a high varroa mite infestation level on bees in September, were weak in September, had a queen older than one year or the beekeeper had few years of beekeeping experience. However, the effect of a high varroa mite infestation level in September had by far the greatest potential to raise the winter losses compared to the other significant factors., Competing Interests: LM, HK, AG, ID and RM are employees of the Austrian Agency for Health and Food Safety Ldt. (AGES). AGES is a limited liability company governed by private law which is wholly owned by the Republic of Austria. Due to the special ownership structure and the lack of self-interest of AGES, the particular working relationship is no circumstance that would call into question the author‘s impartiality or might give rise to a conflict of interest. This does not alter our adherence to PLOS ONE policies on sharing data and materials’.

The health of honey bee colonies is, amongst others, affected by the amount, quality and diversity of available melliferous plants. Since landscape is highly diverse throughout Austria regarding the availability of nutritional resources, we used data from annual surveys on honey bee colony losses ranging over six years to analyse a possible relationship with land use. The data set comprises reports from a total of 6,655 beekeepers and 129,428 wintered honey bee colonies. Regions surrounding the beekeeping operations were assigned to one of six clusters according to their composition of land use categories by use of a hierarchical cluster analysis, allowing a rough distinction between urban regions, regions predominated by semi-natural areas and pastures, and mainly agricultural environments. We ran a Generalised Linear Mixed Model and found winter colony mortality significantly affected by operation size, year, and cluster membership, but also by the interaction of year and cluster membership. Honey bee colonies in regions composed predominantly of semi-natural areas, coniferous forests and pastures had the lowest loss probability in four out of six years, and loss probabilities within these regions were significantly lower in five out of six years compared to those within regions composed predominantly of artificial surfaces, broad-leaved and coniferous forest.

We investigated the importance of protein nutrition for honey bee immunity. Different protein diets (monofloral pollen of Helianthus spp., Sinapis spp., Asparagus spp., Castanea spp., a mixture of the four different pollen and the pollen substitute Feedbee TM ) were fed to honey bees in cages ad libitum . After 18 days of feeding, apidaecin 1 isoforms concentration in the thorax were measured using nanoflow liquid chromatography coupled with mass spectrometry. Expression levels of genes, coding for apidaecins and abaecin in the abdomen were determined using quantitative PCR. The results indicate that protein-containing nutrition in adult worker honey bees can trigger certain metabolic responses. Bees without dietary protein showed lower apidaecin 1 isoforms concentrations. The significantly lowest concentration of apidaecin 1 isoforms was found in the group that was fed no pollen diet when compared to Asparagus , Castanea , Helianthus , and Sinapis pollen or the pollen supplement FeedBee TM . Expression levels of the respective genes were also affected by the protein diets and different expression levels of these two antimicrobial peptides were found. Positive correlation between concentration and gene expression of apidaecins was found. The significance of feeding bees with different protein diets, as well as the importance of pollen nutrition for honey bee immunity is demonstrated.

Multiple infections are common in honey bees, Apis mellifera, but the possible role of nutrition in this regard is poorly understood. Microsporidian infections, which are promoted by protein-fed, can negatively correlate with virus infections, but the role of protein nutrition for the microsporidian-virus interface is unknown. Here, we challenged naturally deformed wing virus - B (DWV-B) infected adult honey bee workers fed with or without pollen ( = protein) in hoarding cages, with the microsporidian Nosema ceranae. Bee mortality was recorded for 14 days and N. ceranae spore loads and DWV-B titers were quantified. Amongst the groups inoculated with N. ceranae, more spores were counted in protein-fed bees. However, N. ceranae infected bees without protein-diet had reduced longevity compared to all other groups. N. ceranae infection had no effect on protein-fed bee's longevity, whereas bees supplied only with sugar-water showed reduced survival. Our data also support that protein-feeding can have a significant negative impact on virus infections in insects. The negative correlation between N. ceranae spore loads and DWV-B titers was stronger expressed in protein-fed hosts. Proteins not only enhance survival of infected hosts, but also significantly shape the microsporidian-virus interface, probably due to increased spore production and enhanced host immunity.

Recently, evidence has shown that variations in the cuticular hydrocarbons (CHCs) profile allow healthy honeybees to identify diseased nestmates, eliciting agonistic responses in the former. Here, we determined whether these 'immunologic cues' emitted by diseased nestmates were only detected by workers, who consequently took hygienic measures and excluded these individuals from the colony, or whether queens were also able to detect these cues and respond accordingly. Healthy honeybee queens were exposed to (i) healthy, (ii) Ringer-injected and (iii) lipopolysaccharide (LPS)-injected nestmates by allowing direct body contact. Quantitative differences in the CHC profiles of these three groups were measured using GC-MS. The transcript levels of the products of four genes that encode for antimicrobial peptides (AMPs), which are part of the queen's immune response, were measured in bees exposed to direct contact using qPCR. A significant increase in the transcript levels of these AMP genes over baseline levels in queens was observed when body contact was allowed between the queens and the Ringer- and LPS-injected nestmates. These results provide the first evidence that the detection of CHCs contributes to the initiation of an immune response in insects. In an additional experiment, CHCs were extracted from diseased workers and directly presented to queens, which also evoked a similar immune response. A potential mechanism that relied on volatile compounds could be ruled out by conducting a distance experiment. The study helps to expand our knowledge of chemical communication in insects and sheds light on a likely new mechanism of social immunity., (© 2017 The Authors Molecular Ecology Published by John Wiley & Sons Ltd.)

We measured the distribution of sugar solution within groups of caged honey bees (Apis mellifera) under standard in vitro laboratory conditions using 14C polyethylene glycol as a radioactive marker to analyze ingestion by individual bees after group feeding. We studied the impact of different experimental setups by varying the number of bees, age of bees, origin of bees, duration of experiment, the amount of available diet, and the influence of the neurotoxic pesticide imidacloprid in the diet on the feeding and food sharing behavior (trophallaxis). Sugar solution was non-uniformly distributed in bees in 36 out of 135 cages. As a measure of the extent to which the sugar diet was equally distributed between caged bees, we calculated the (inner 80%) intake ratio by dividing the intake of the 90th percentile bee by the intake of the 10th percentile bee. This intake ratio ranged from 1.3 to 94.8 in 133 individual cages, further supporting a non-uniform distribution of food among caged bees. We can expect a cage with 10 or 30 bees containing one bee that ingests, on average, the 8.8-fold of the bee in the same cage ingesting the smallest quantity of food. Inner 80% intake ratios were lower in experiments with a permanent or chronic offering of labelled sugar solution compared to temporary or acute feedings. After pooling the data of replicates to achieve a higher statistical power we compared different experimental setups. We found that uniform food distribution is best approached with 10 newly emerged bees per cage, which originate from a brood comb from a single colony. We also investigated the trophallaxis between caged honey bees which originally consumed the diet and newly added bees. Color marked bees were starved and added to the cages in a ratio of 10:5 or 20:20 after the initial set of bees consumed all the labelled sugar solution. The distribution of the labelled sugar solution by trophallaxis within 48 hours to added bees was 25% (10:5) or 45% (20:20) of the initial sugar solution. Imidacloprid at its median lethal dose (LD50) in the sugar solution reduced this post-feeding food transmission to 27% (20:20). Our results show that differences in food intake exist within caged bees that may lead to differential exposure that can influence the interpretation of toxicity tests.

Insect pollinators are essential to global food production. For this reason, it is alarming that honey bee (Apis mellifera) populations across the world have recently seen increased rates of mortality. These changes in colony mortality are often ascribed to one or more factors including parasites, diseases, pesticides, nutrition, habitat dynamics, weather and/or climate. However, the effect of climate on colony mortality has never been demonstrated. Therefore, in this study, we focus on longer-term weather conditions and/or climate's influence on honey bee winter mortality rates across Austria. Statistical correlations between monthly climate variables and winter mortality rates were investigated. Our results indicate that warmer and drier weather conditions in the preceding year were accompanied by increased winter mortality. We subsequently built a statistical model to predict colony mortality using temperature and precipitation data as predictors. Our model reduces the mean absolute error between predicted and observed colony mortalities by 9% and is statistically significant at the 99.9% confidence level. This is the first study to show clear evidence of a link between climate variability and honey bee winter mortality., (Copyright © 2016 British Geological Survey, NERC. Published by Elsevier B.V. All rights reserved.)

Disclosing interactions between pesticides and bee infections is of most interest to understand challenges that pollinators are facing and to which extent bee health is compromised. Here, we address the individual and combined effect that three different pesticides (dimethoate, clothianidin and fluvalinate) and an American foulbrood (AFB) infection have on mortality and the cellular immune response of honeybee larvae. We demonstrate for the first time a synergistic interaction when larvae are exposed to sublethal doses of dimethoate or clothianidin in combination with Paenibacillus larvae, the causative agent of AFB. A significantly higher mortality than the expected sum of the effects of each individual stressor was observed in co-exposed larvae, which was in parallel with a drastic reduction of the total and differential hemocyte counts. Our results underline that characterizing the cellular response of larvae to individual and combined stressors allows unmasking previously undetected sublethal effects of pesticides in colony health., Competing Interests: The authors declare no competing financial interests.

Self-structuring patterns can be observed all over the universe, from galaxies to molecules to living matter, yet their emergence is waiting for full understanding. We discovered a simple motion law for moving and interacting self-propelled particles leading to a self-structuring, self-reproducing and self-sustaining life-like system. The patterns emerging within this system resemble patterns found in living organisms. The emergent cells we found show a distinct life cycle and even create their own ecosystem from scratch. These structures grow and reproduce on their own, show self-driven behavior and interact with each other. Here we analyze the macroscopic properties of the emerging ecology, as well as the microscopic properties of the mechanism that leads to it. Basic properties of the emerging structures (size distributions, longevity) are analyzed as well as their resilience against sensor or actuation noise. Finally, we explore parameter space for potential other candidates of life. The generality and simplicity of the motion law provokes the thought that one fundamental rule, described by one simple equation yields various structures in nature: it may work on different time- and size scales, ranging from the self-structuring universe, to emergence of living beings, down to the emergent subatomic formation of matter.

Honeybee (Apis mellifera) imagines are resistant to the Gram-positive bacterium Paenibacillus larvae (P. larvae), causative agent of American foulbrood (AFB), whereas honeybee larvae show susceptibility against this pathogen only during the first 48 h of their life. It is known that midgut homogenate of adult honeybees as well as a homogenate of aged larvae exhibit strong anti-P. larvae activity. A bioactivity-guided LC-HRMS analysis of midgut homogenate resulted in the identification of 1-oleoyl-sn-glycero-3-phosphocholine (LPC) pointing to a yet unknown immune defence in adult honeybees against P. larvae. Antimicrobial activity of LPC was also demonstrated against Melissococcus plutonius, causative agent of European Foulbrood. To demonstrate an AFB-preventive effect of LPC in larvae, artificially reared larvae were supplemented with LPC to evaluate its toxicity and to assess whether, after infection with P. larvae spores, LPC supplementation prevents AFB infection. 10 μg LPC per larva applied for 3 d significantly lowered mortality due to AFB in comparison to controls. A potential delivery route of LPC to the larvae in a colony via nurse bees was assessed through a tracking experiment using fluorescent-labelled LPC. This yet undescribed and non-proteinous defense of honeybees against P. larvae may offer new perspectives for a treatment of AFB without the utilization of classic antibiotics.

Hydroxymethylfurfural (HMF) is a heat-formed, acid-catalyzed contaminant of sugar syrups, which find their way into honey bee feeding. As HMF was noted to be toxic to adult honey bees, we investigated the toxicity of HMF towards larvae. Therefore we exposed artificially reared larvae to a chronic HMF intoxication over 6 days using 6 different concentrations (5, 50, 750, 5000, 7500 and 10,000 ppm) and a control. The mortality was assessed from day 2 to day 7 (d7) and on day 22 (d22). Concentrations ranging from 5 to 750 ppm HMF did not show any influence on larval or pupal mortality compared to controls (p > 0.05; Kaplan-Meier analysis). Concentrations of 7500 ppm or higher caused a larval mortality of 100%. An experimental LC50 of 4280 ppm (d7) and 2424 ppm (d22) was determined. The calculated LD50 was 778 µg HMF per larva on d7 and 441 µg HMF on d22. Additionally, we exposed adult honey bees to high concentrations of HMF to compare the mortality to the results from larvae. On d7 larvae are much more sensitive against HMF than adult honey bees after 6 days of feeding. However, on d22 after emergence adults show a lower LC50, which indicates a higher sensitivity than larvae. As toxicity of HMF against honey bees is a function of time and concentration, our results indicate that HMF in supplemental food will probably not cause great brood losses. Yet sublethal effects might decrease fitness of the colony.

The long-term effects of early adverse experiences on later psychosocial functioning are well described in humans, but sparsely documented for chimpanzees. In our earlier studies, we investigated the effects of maternal and social deprivation on three groups of ex-laboratory chimpanzees who experienced either an early or later onset of long-term deprivation. Here we expand our research by adding data on subjects that came from two stable zoo groups. The groups comprised of early maternally deprived wild-caught chimpanzees and non-deprived zoo-born chimpanzees. We found that compared to zoo chimpanzees, ex-laboratory chimpanzees were more restricted regarding their association partners in the newly formed groups, but not during their second year of group-life, indicating that social stability has an important influence on the toleration of association partners close-by. Social grooming activity, however, was impaired in early long-term deprived ex-laboratory chimpanzees as well as in early maternally deprived zoo chimpanzees compared to non-deprived zoo chimpanzees. Thus, we conclude that early maternal loss has lifelong effects on the social integration of chimpanzees which becomes evident in their grooming networks. Although the retrospective nature of our study prevents a clear causal explanation, our results are of importance for understanding the development of social competence in chimpanzees.

Some parasites of social insects are able to exploit the exchange of food between nestmates via trophallaxis, because they are chemically disguised as nestmates. However, a few parasites succeed in trophallactic solicitation although they are attacked by workers. The underlying mechanisms are not well understood. The small hive beetle (=SHB), Aethina tumida, is such a parasite of honey bee, Apis mellifera, colonies and is able to induce trophallaxis. Here, we investigate whether SHB trophallactic solicitation is innate and affected by sex and experience. We quantified characteristics of the trophallactic solicitation in SHBs from laboratory-reared individuals that were either bee-naïve or had 5 days experience. The data clearly show that SHB trophallactic solicitation is innate and further suggest that it can be influenced by both experience and sex. Inexperienced SHB males begged more often than any of the other groups had longer breaks than their experienced counterparts and a longer soliciting duration than both experienced SHB males and females, suggesting that they start rather slowly and gain more from experience. Successful experienced females and males were not significantly different from each other in relation to successful trophallactic interactions, but had a significantly shorter soliciting duration compared to all other groups, except successful inexperienced females. Trophallactic solicitation success, feeding duration and begging duration were not significantly affected by either SHB sex or experience, supporting the notion that these behaviors are important for survival in host colonies. Overall, success seems to be governed by quality rather than quantity of interactions, thereby probably limiting both SHB energy investment and chance of injury (<1%). Trophallactic solicitation by SHBs is a singular example for an alternative strategy to exploit insect societies without requiring chemical disguise. Hit-and-run trophallaxis is an attractive test system to get an insight into trophallaxis in the social insects.

In this paper, a distributed adaptive partitioning algorithm inspired by division of labor in honeybees is investigated for its applicability in a swarm of underwater robots in one hand and is qualitatively compared with the behavior of honeybee colonies on the other hand. The algorithm, partitioning social inhibition (PSI), is based on local interactions and uses a simple logic inspired from age-polyethism and task allocation in honeybee colonies. The algorithm is analyzed in simulation and is successfully applied here to partition a swarm of underwater robots into groups demonstrating its adaptivity to changes and applicability in real world systems. In a turn towards the inspiration origins of the algorithm, three honeybee colonies are then studied for age-polyethism behaviors and the results are contrasted with a simulated swarm running the PSI algorithm. Similar effects are detected in both the biological and simulated swarms suggesting biological plausibility of the mechanisms employed by the artificial system.

Immune responses of invertebrates imply more than developing a merely unspecific response to an infection. Great interest has been raised to unveil whether this investment into immunity also involves fitness costs associated to the individual or the group. Focusing on the immune responses of honeybees, we use the well-studied insect bumblebee for comparison. Bumblebees are capable of producing specific immune responses to infections whereas this has not been assessed for honeybees so far. We investigated whether a prior bacterial encounter provides protection against a later exposure to the same or a different bacterium in honeybees. Additionally, we studied whether the foraging activities of honeybees and bumblebees are affected upon immune stimulation by assessing the flight performance. Finally, the acceptance behavior of nestmates toward immune-challenged honeybees was determined. Results show that despite stimulating the immune system of honeybees, no protective effects to infections were found. Further, honeybees were not affected by an immune challenge in their flight performance whereas bumblebees showed significant flight impairment. Immune-challenged honeybees showed lower survival rates than naive individuals when introduced into a regular colony. Here, we reveal different immune response-cost scenarios in honeybees and bumblebees for the first time.

A grand challenge in the field of artificial life is to find a general theory of emergent self-organizing systems. In swarm systems most of the observed complexity is based on motion of simple entities. Similarly, statistical mechanics focuses on collective properties induced by the motion of many interacting particles. In this article we apply methods from statistical mechanics to swarm systems. We try to explain the emergent behavior of a simulated swarm by applying methods based on the fluctuation theorem. Empirical results indicate that swarms are able to produce negative entropy within an isolated subsystem due to frozen accidents. Individuals of a swarm are able to locally detect fluctuations of the global entropy measure and store them, if they are negative entropy productions. By accumulating these stored fluctuations over time the swarm as a whole is producing negative entropy and the system ends up in an ordered state. We claim that this indicates the existence of an inverted fluctuation theorem for emergent self-organizing dissipative systems. This approach bears the potential of general applicability.

Background: Sensitization to Hymenoptera venom without systemic sting reactions (SSRs) is commonly observed in the general population. Clinical relevance for a future sting has not yet been investigated., Objective: We aimed to evaluate the effect of these debatable sensitizations with deliberate sting challenges and to monitor serologic changes for up to 2 years., Methods: One hundred thirty-one challenges with bees and wasps were performed in 94 subjects with a hitherto irrelevant sensitization. The clinical outcome was recorded, and results of specific IgE (sIgE) determinations, skin tests, and basophil activation tests were correlated to the sting reaction. sIgE levels were monitored in reactors and nonreactors after 3 hours, 1 week, 4 weeks, and 1 year., Results: Only 5 (5.3%) patients had SSRs, but 41 (43.6%) had large local reactions (LLRs) after the sting. Compared with the general population, there was a 9.5-fold higher risk for LLRs but not for SSRs. Three hours after the sting, sIgE levels slightly decreased, but none of the 94 subjects' results turned negative. After 1 week, sIgE levels already increased, increasing up to 3.5-fold (range, 0.2- to 34.0-fold) baseline levels after 4 weeks. To assess the clinical relevance of this increase, we randomly selected 18 patients for a re-sting. Again, 50% had an LLR, but none had an SSR., Conclusion: Although sensitization to Hymenoptera venoms was common, the risk of SSRs in sensitized subjects was low in our study. The sIgE level increase after the sting was not an indicator for conversion into symptomatic sensitization. Currently available tests were not able to distinguish between asymptomatic sensitization, LLRs, and SSRs., (Copyright © 2013 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.)

The in vitro inhibitory potential of 50 extracts from various species of the flowering plant genus Hypericum was investigated using the Kirby-Bauer disk diffusion susceptibility test against Paenibacillus larvae, a spore-forming, Gram-positive bacterial pathogen that causes American foulbrood (AFB), a lethal disease affecting honeybee brood worldwide. Of the tested extracts, 14 were identified as highly active against P. larvae as compared to the activity of the positive control, indicating the presence of highly potent antibacterial compounds in the extracts. Examination of these extracts using TLC and HPLC/MS analyses revealed the presence of acylphloroglucinol and filicinic-acid derivatives. Six pure compounds isolated from these extracts, viz., hyperforin (1), uliginosin B (2), uliginosin A (3), 7-epiclusianone (4), albaspidin AA (5), and drummondin E (6), displayed strong antibacterial activity against the vegetative form of P. larvae (MIC ranging from 0.168-220 μM). Incubation of P. larvae spores with the lipophilic extract of Hypericum perforatum and its main acylphloroglucinol constituent 1 led to the observation of significantly fewer colony forming units as compared to the negative control, indicating that the acylphloroglucinol scaffold represents an interesting lead structure for the development of new AFB control agents., (Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.)

Maternal immune experience acquired during pathogen exposure and passed on to progeny to enhance resistance to infection is called trans-generational immune priming (TgIP). In eusocial insects like honeybees, TgIP would result in a significant improvement of health at individual and colony level. Demonstrated in invertebrates other than honeybees, TgIP has not yet been fully elucidated in terms of intensity and molecular mechanisms underlying this response. Here, we immune-stimulated honeybee queens with Paenibacillus larvae (Pl), a spore-forming bacterium causing American Foulbrood, the most deadly bee brood disease worldwide. Subsequently, offspring of stimulated queens were exposed to spores of Pl and mortality rates were measured to evaluate maternal transfer of immunity. Our data substantiate the existence of TgIP effects in honeybees by direct evaluation of offspring resistance to bacterial infection. A further aspect of this study was to investigate a potential correlation between immune priming responses and prohaemocytes-haemocyte differentiation processes in larvae. The results point out that a priming effect triggers differentiation of prohaemocytes to haemocytes. However, the mechanisms underlying TgIP responses are still elusive and require future investigation.

A computer program that tracks animal behavior, thereby revealing various features and mechanisms of social animals, is a powerful tool in ethological research. Because honeybee colonies are populated by thousands of bees, individuals co-exist in high physical densities and are difficult to track unless specifically tagged, which can affect behavior. In addition, honeybees react to light and recordings must be made under special red-light conditions, which the eyes of bees perceive as darkness. The resulting video images are scarcely distinguishable. We have developed a new algorithm, K-Track, for tracking numerous bees in a flat laboratory arena. Our program implements three main processes: (A) The object (bee's) region is detected by simple threshold processing on gray scale images, (B) Individuals are identified by size, shape and spatiotemporal positional changes, and (C) Centers of mass of identified individuals are connected through all movie frames to yield individual behavioral trajectories. The tracking performance of our software was evaluated on movies of mobile multi-artificial agents and of 16 bees walking around a circular arena. K-Track accurately traced the trajectories of both artificial agents and bees. In the latter case, K-track outperformed Ctrax, well-known software for tracking multiple animals. To investigate interaction events in detail, we manually identified five interaction categories; 'crossing', 'touching', 'passing', 'overlapping' and 'waiting', and examined the extent to which the models accurately identified these categories from bee's interactions. All 7 identified failures occurred near a wall at the outer edge of the arena. Finally, K-Track and Ctrax successfully tracked 77 and 60 of 84 recorded interactive events, respectively. K-Track identified multiple bees on a flat surface and tracked their speed changes and encounters with other bees, with good performance.

Endothermic heat production is a crucial evolutionary adaptation that is, amongst others, responsible for the great success of honeybees. Endothermy ensures the survival of the colonies in harsh environments and is involved in the maintenance of the brood nest temperature, which is fundamental for the breeding and further development of healthy individuals and thus the foraging and reproduction success of this species. Freshly emerged honeybees are not yet able to produce heat endothermically and thus developed behavioural patterns that result in the location of these young bees within the warm brood nest where they further develop and perform tasks for the colony. Previous studies showed that groups of young ectothermic honeybees exposed to a temperature gradient collectively aggregate at the optimal place with their preferred temperature of 36 °C but most single bees do not locate themselves at the optimum. In this work we further investigate the behavioural patterns that lead to this collective thermotaxis. We tested single and groups of young bees concerning their ability to discriminate a local from a global temperature optimum and, for groups of bees, analysed the speed of the decision making process as well as density dependent effects by varying group sizes. We found that the majority of tested single bees do not locate themselves at the optimum whereas sufficiently large groups of bees are able to collectively discriminate a suboptimal temperature spot and aggregate at 36 °C. Larger groups decide faster than smaller ones, but in larger groups a higher percentage of bees may switch to the sub-optimum due to crowding effects. We show that the collective thermotaxis is a simple but well evolved, scalable and robust social behaviour that enables the collective of bees to perform complex tasks despite the limited abilities of each individual.

Swarm systems are based on intermediate connectivity between individuals and dynamic neighborhoods. In natural swarms self-organizing principles bring their agents to that favorable level of connectivity. They serve as interesting sources of inspiration for control algorithms in swarm robotics on the one hand, and in modular robotics on the other hand. In this paper we demonstrate and compare a set of bio-inspired algorithms that are used to control the collective behavior of swarms and modular systems: BEECLUST, AHHS (hormone controllers), FGRN (fractal genetic regulatory networks), and VE (virtual embryogenesis). We demonstrate how such bio-inspired control paradigms bring their host systems to a level of intermediate connectivity, what delivers sufficient robustness to these systems for collective decentralized control. In parallel, these algorithms allow sufficient volatility of shared information within these systems to help preventing local optima and deadlock situations, this way keeping those systems flexible and adaptive in dynamic non-deterministic environments.

Adverse rearing conditions are considered a major factor in the development of abnormal behavior. We investigated the overall levels, the prevalence and the diversity of abnormal behavior of 18 adult former laboratory chimpanzees, who spent about 20 years single caged, over a two-year period following re-socialization. According to the onset of deprivation, the individuals were classified as early deprived (EDs, mean: 1.2 years) or late deprived (LDs, mean: 3.6 years). The results are based on 187.5 hours of scan sampling distributed over three sample periods: subsequent to re-socialization and during the first and second year of group-living. While the overall levels and the diversity of abnormal behavior remained stable over time in this study population, the amplifying effects of age at onset of deprivation became apparent as the overall levels of abnormal behavior of EDs were far above those of LDs in the first and second year of group-living, but not immediately after re-socialization. The most prevalent abnormal behaviors, including eating disorders and self-directed behaviors, however, varied in their occurrence within subjects across the periods. Most important, the significance of social companionship became obvious as the most severe forms of abnormal behavior, such as dissociative and self-injurious behaviors declined.

A honey bee queen mates with a number of drones a few days after she emerges as an adult. Spermatozoa of different drones are stored in her spermatheca and used for the rest of the queen's life to fertilize eggs. Sperm usage is thought to be random, so that the patriline distribution within a honey bee colony would remain constant over time. In this study we assigned the progeny of a naturally mated honey bee queen to patrilines using microsatellite markers at the queen's age of two, three and four years. No significant changes in patriline distribution occurred within each of two foraging seasons, with samples taken one and five months apart, respectively. Overall and pair-wise comparisons between the three analyzed years reached significant levels. Over the three-year period we found a trend for patrilines to become more equally represented with time. It is important to note that this study was performed with a single queen, and thus individual and population variation in sperm usage patterns must be assessed. We discuss long-term changes in patriline composition due to mixing processes in the queen's spermatheca, following incomplete mixing of different drones' sperm after mating.

One of the main challenges in automatic controller synthesis is to develop methods that can successfully be applied for complex tasks. The difficulty is increased even more in the case of settings with multiple interacting agents. We apply the artificial homeostatic hormone system (AHHS) approach, which is inspired by the signaling network of unicellular organisms, to control a system of several independently acting agents decentrally. The approach is designed for evaluation-minimal, artificial evolution in order to be applicable to complex modular robotics scenarios. The performance of AHHS controllers is compared with neuroevolution of augmenting topologies (NEAT) in the coupled inverted pendulums benchmark. AHHS controllers are found to be better for multimodular settings. We analyze the evolved controllers with regard to the usage of sensory inputs and the emerging oscillations, and we give a nonlinear dynamics interpretation. The generalization of evolved controllers to initial conditions far from the original conditions is investigated and found to be good. Similarly, the performance of controllers scales well even with module numbers different from the original domain the controller was evolved for. Two reference implementations of a similar controller approach are reported and shown to have shortcomings. We discuss the related work and conclude by summarizing the main contributions of our work.

Early social deprivation in highly social mammals interferes with their varying needs for security and stimulation. Toleration of social stimulation was studied in 18 adult ex-laboratory chimpanzees, who had been deprived for 16 to 27 years, during their 1st year after resocialization into 1 of 3 social groups. For this, a model of social competence was developed with 5 grades of social stimulation. The chimpanzees were classified as either early deprived (EDs; M = 1.2 years) or late deprived (LDs; M = 3.6 years) according to their age at entering the laboratory. EDs did not differ significantly from LD chimpanzees in the first 3 grades of social stimulation (i.e., the toleration of stationary vicinity, initiation of brief sociopositive contacts and gentle social play). LDs, however, clearly exceeded EDs in the 4th and 5th grade of social stimulation (i.e., their engagement in allogrooming and their toleration of passive close proximity). Furthermore, LDs showed greater social initiative in changing a social situation and had more expanded grooming networks compared with EDs. Moreover, in LDs and ED females, but not in ED males, toleration of stationary vicinity increased from the 1st to the 2nd year of group living.

Background: Double sensitization (DS) to bee and vespid venom is frequently observed in the diagnosis of hymenoptera venom allergy, but clinically relevant DS is rare. Therefore it is sophisticated to choose the relevant venom for specific immunotherapy and overtreatment with both venoms may occur. We aimed to compare currently available routine diagnostic tests as well as experimental tests to identify the most accurate diagnostic tool., Methods: 117 patients with a history of a bee or vespid allergy were included in the study. Initially, IgE determination by the ImmunoCAP, by the Immulite, and by the ADVIA Centaur, as well as the intradermal test (IDT) and the basophil activation test (BAT) were performed. In 72 CAP double positive patients, individual IgE patterns were determined by western blot inhibition and component resolved diagnosis (CRD) with rApi m 1, nVes v 1, and nVes v 5., Results: Among 117 patients, DS was observed in 63.7% by the Immulite, in 61.5% by the CAP, in 47.9% by the IDT, in 20.5% by the ADVIA, and in 17.1% by the BAT. In CAP double positive patients, western blot inhibition revealed CCD-based DS in 50.8%, and the CRD showed 41.7% of patients with true DS. Generally, agreement between the tests was only fair and inconsistent results were common., Conclusion: BAT, CRD, and ADVIA showed a low rate of DS. However, the rate of DS is higher than expected by personal history, indicating that the matter of clinical relevance is still not solved even by novel tests. Furthermore, the lack of agreement between these tests makes it difficult to distinguish between bee and vespid venom allergy. At present, no routinely employed test can be regarded as gold standard to find the clinically relevant sensitization.

Elucidating the age-dependent alterations in transgenic (Tg) mice overexpressing amyloid-β protein precursor (AβPP) is important for understanding the pathogenesis of Alzheimer's disease (AD) and designing experimental therapies. Cross-studies have previously characterized some time-dependent behavioral and pathological alterations in AβPP Tg mice, however, a more comprehensive longitudinal study is needed to fully examine the progressive nature of behavioral deficits in these mice. In order to better understand the age- and gender-dependent progression of behavioral alterations, we performed a longitudinal study wherein Tg mice overexpressing human AβPP751 with the London (V717I) and Swedish (K670M/N671L) mutations under the regulatory control of the neuron specific murine (m)Thy-1 promoter (mThy1-hAβPP751) were behaviorally analyzed at 3 months and then re-tested at 6 and 9 months of age. The results show that there was an age-associated impairment in learning in the water maze task and habituation in the hole-board task. Motor coordination of the mThy1-hAβPP751 Tg mice was well-preserved throughout the investigated life span however, gender-specific deficits were observed in spontaneous activity and thigmotaxis. Neuropathologically, mThy1-hAβPP751 Tg mice displayed a progressive increase in the number of Aβ plaques and mean plaque size in the cortex and hippocampus from 3 to 6 and from 6 to 9 months of age. Taken together, these results indicate that the mThy1-hAβPP751 Tg mice model AD from the early onset of the disease through to later stages, allowing them to be utilized at numerous points during the timeline for drug test designs.

Maternal or social deprivation during early infancy inevitably produces social deficiencies in juvenile chimpanzees. Hypothesizing such deficiencies to persist into adulthood (a), and, as in humans, a sensitive period in early infancy for attachment formation (b), we predicted and found behavioral differences in resocialized adult ex-laboratory chimpanzees after about 20 years of solitary confinement depending on their age at onset of deprivation: early deprived (ED; mean: 1.2 years) chimpanzees engaged significantly less in social interactions, spent less time associated, and showed more nonsocial idiosyncrasies than did late deprived (LD; mean: 3.6 years) chimpanzees. In addition to these individual attributes relational qualities, specifically the combination of ED and LD chimpanzees within social groups, have an impact on social recovery. LDs can best exploit their social potential in the company of other LDs and EDs tend to stagnate in their recovery when socialized with other EDs.

The relation between the respiratory activity of resting honeybees and ambient temperature (T(a)) was investigated in the range of 5-40 degrees C. Bees were kept in a temperature controlled flow through respirometer chamber where their locomotor and endothermic activity, as well as abdominal ventilatory movements was recorded by infrared thermography. Surprisingly, true resting bees were often weakly endothermic (thorax surface up to 2.8 degrees C warmer than abdomen) at a T(a) of 14-30 degrees C. Above 33 degrees C many bees cooled their body via evaporation from their mouthparts. A novel mathematical model allows description of the relationship of resting (standard) metabolic rate and temperature across the entire functional temperature range of bees. In chill coma (<11 degrees C) bees were ectothermic and CO(2) release was mostly continuous. CO(2) release rate (nls(-1)) decreased from 9.3 at 9.7 degrees C to 5.4 at 5 degrees C. At a T(a) of >11 degrees C CO(2) was released discontinuously. In the bees' active temperature range mean CO(2) production rate (nls(-1)) increased sigmoidally (10.6 at 14.1 degrees C, 24.1 at 26.5 degrees C, and 55.2 at 38.1 degrees C), coming to a halt towards the upper lethal temperature. This was primarily accomplished by an exponential increase in gas exchange frequency (0.54 and 3.1 breaths min(-1) at 14.1 and 38.1 degrees C) but not in released CO(2) volume per respiratory cycle (1487 and 1083 nl cycle(-1) at 14.1 and 38.1 degrees C). Emission of CO(2) bursts was mostly (98%) accompanied by abdominal ventilation movements even in small CO(2) bursts. Larger bursts coincided with a longer duration of active ventilation. An increased amount of CO(2) expelled per unit time of ventilation indicates a higher efficiency of ventilation at high ambient temperatures.

Expression of prion protein has been reported for a variety of cell types including neuronal cells, haematopoietic stem cells, lymphocytes, fibroblasts, and epithelial cells. However, the characterization of the physiological roles exhibited by this protein is still in progress and multiple biological functions have been described to date. In this study we have characterized the contribution of prion protein during hormone-induced differentiation of mouse mammary gland epithelial cells. We present evidence that prion expression enhances the differentiation-capabilities of these cells indicating novel physiological roles during mammary gland development. In addition we were able to demonstrate the presence of prion molecules resistant to mild proteinase digestion in differentiated mammary gland epithelial cells. This represents the first report of proteinase-resistant prion proteins in a physiological, non-pathogenic context.

Butterflies of the genus Heliconius are well known for their peculiar habits of utilizing pollen as a source of amino acids. Saliva plays a major role in the process of extracting amino acids and proteins from the pollen grains. In this investigation, we obtained samples of saliva from adult Heliconius melpomene by placing pumpkin pollen or fine glass-beads on the proboscis, which stimulates the butterflies to release saliva. Proteolytic activity was determined in the saliva by an insoluble protein-dye that turns blue when cleaved by proteases. Its extinction value was measured with a spectrophotometer at 595 nm. Both the saliva sampled with pollen and the saliva obtained from inert glass-beads exhibit proteolytic activity demonstrating that the saliva contains proteases. The proteolytic activity of the pollen/saliva samples was higher than that of the glass-bead/saliva samples, which we attribute to the stimulating effects of pollen, such as taste, smell, and texture, and not to proteases which might have been liberated from the pollen. This is indicated by the fact that pollen samples without saliva showed only a negligible indication for proteolytic activity. In general, females exhibit higher proteolytic activities than males, presumably due to their greater amino acid investment in reproduction. We present here first evidence for the existence of proteases in the saliva of a butterfly species and suggest that these enzymes are crucial for the use of amino acids and proteins from pollen in Heliconius butterflies.

Honeybee workers, Apis mellifera, can reproduce in queenless colonies. The production of queen-like pheromones may be associated with their reproductive activity and induce nestmates to respond by feeding them. Such frequent trophallaxis could supply their protein needs for oogenesis, constituting a social pathway to worker reproduction. However, some individuals can develop ovaries without producing queen pheromones. The consumption of protein-rich pollen could be an alternative solitary pathway for them to satisfy this dietary requirement. In order to investigate the way in which workers obtain proteins for oogenesis, we created orphaned worker groups and determined ovarian and pheromonal development in relation to pollen consumption of selected workers. Individuals that did not consume pollen had significantly more developed ovaries and produced significantly more queen mandibular pheromone than workers that fed directly on pollen. Our results suggest that workers producing queen-like secretions are fed trophallactically. However, reproductive workers that lacked queen pheromones had consumed little or no pollen, suggesting that they also obtained trophallaxis. Although pollen consumption might contribute to sustaining oogenesis, it does not appear to be sufficient. Trophallaxis as a means of obtaining proteins seems to be necessary to attain reproductive status in queenless honeybee colonies.

The CO(2) production of individual larvae of Apis mellifera carnica, which were incubated within their cells at a natural air humidity of 60-80%, was determined by an open-flow gas analyzer in relation to larval age and ambient temperature. In larvae incubated at 34 degrees C the amount of CO(2) produced appeared to fall only moderately from 3.89 +/- 1.57 microl mg(-1) h(-1) in 0.5-day-old larvae to 2.98 +/- 0.57 microl mg(-1) h(-1) in 3.5-day-old larvae. The decline was steeper up to an age of 5.5 days (0.95 +/- 1.15 microl mg(-1) h(-1)). Our measurements show that the respiration and energy turnover of larvae younger than about 80 h is considerably lower (up to 35%) than expected from extrapolations of data determined in older larvae. The temperature dependency of CO(2) production was determined in 3.5-day-old larvae, which were incubated at temperatures varying from 18 to 38 degrees C in steps of 4 degrees C. The larvae generated 0.48+/-0.03 microl mg(-1) h(-1) CO(2) at 18 degrees C, and 3.97 +/- 0.50 microl mg(-1) h(-1) CO(2) at 38 degrees C. The temperature-dependent respiration rate was fitted to a logistic curve. We found that the inflection point of this curve (32.5 degrees C) is below the normal brood nest temperature (33-36 degrees C). The average Q(10) was 3.13, which is higher than in freshly emerged resting honeybees but similar to adult bees. This strong temperature dependency enables the bees to speed up brood development by achieving high temperatures. On the other hand, the results suggest that the strong temperature dependency forces the bees to maintain thermal homeostasis of the brood nest to avoid delayed brood development during periods of low temperature.

Weight cycling may cause a redistribution of body fat to the upper body fat compartments. We investigated the distribution of subcutaneous adipose tissue (SAT) in 30 overweight women with a history of weight-cycling and age-matched controls (167 normal weight and 97 overweight subjects). Measurements of SAT were performed using an optical device, the Lipometer. The SAT topography describes the thicknesses of SAT layers at 15 anatomically well-defined body sites from neck to calf. The overweight women with a history of weight cycling had significantly thicker SAT layers on the upper body compared to the overweight controls, but even thinner SAT layers on their legs than the normal weight women. An android fat pattern was attributed to overweight females and, even more pronounced, to the weight cyclers. The majority of normal weight women showed a gynoid fat pattern. Using stepwise discriminant analysis, 89.0% of all weight cyclers and overweight controls could be classified correctly into the two groups. These findings show the importance of normal weight maintenance as a health-promoting factor.

The new optical device, the lipometer, enables the noninvasive, quick, safe, and precise determination of the thickness of subcutaneous adipose tissue (SAT) layers at any given site of the human body. Fifteen anatomically well-defined body sites from neck to calf describe a SAT topography (SAT-Top) like an individual "fingerprint" of a subject. This SAT-Top was examined in 16 women with polycystic ovary syndrome (PCOS) and compared to the body fat distribution of 87 age-matched healthy controls and 20 type-2 diabetic women. SAT-Top differences of these three groups were described and, to render the possibility of visual comparison, the 15-dimensional body fat information was condensed to a two-dimensional factor plot by factor analysis. All PCOS patients had an android body fat distribution with significantly thinner SAT layers on the legs as compared to healthy controls. Moreover, a hierarchical cluster analysis resulted in two distinctly different groups of PCOS women, a lean (PCOSL) and an obese (PCOSO) cluster: compared to healthy women, lean PCOS patients had significantly lower total SAT development, even though height, weight, and body mass index did not deviate significantly. Especially on the legs, their SAT layers were significantly lowered, indicating a more "apple-like" fat distribution type. Obese PCOS women showed a SAT-Top pattern very similar to that of women with type-2 diabetes, although the mean age difference between these groups was more than 30 years. Compared to healthy controls, the SAT-Top of these obese PCOS patients was strongly shifted into the android direction, appearing as "super-apples" with a significantly increased upper trunk obesity to 237.8% and a significantly decreased leg SAT development to 79.8%., (Copyright 2004 Wiley-Liss, Inc.)

Vagal afferents play a role in gut-brain signaling of physiological and pathological stimuli. Here, we investigated how backdiffusion of luminal HCl or NH(4)OH and pentagastrin-stimulated acid secretion interact in the communication between rat stomach and brain stem. Rats were pretreated intraperitoneally with vehicle or appropriate doses of cimetidine, omeprazole, pentagastrin, dexloxiglumide (CCK(1) receptor antagonist), and itriglumide (CCK(2) receptor antagonist) before intragastric administration of saline or backdiffusing concentrations of HCl or NH(4)OH. Two hours later, neuronal activation in the nucleus of the solitary tract (NTS) and area postrema was visualized by c-Fos immunohistochemistry. Exposure of the rat gastric mucosa to HCl (0.15-0.5 M) or NH(4)OH (0.1-0.3 M) led to a concentration-dependent expression of c-Fos in the NTS, which was not related to gender, gastric mucosal injury, or gastropyloric motor alterations. The c-Fos response to HCl was diminished by cimetidine and omeprazole, enhanced by pentagastrin, and left unchanged by dexloxiglumide and itriglumide. Pentagastrin alone caused an omeprazole-resistant expression of c-fos, which in the NTS was attenuated by itriglumide and prevented by dexloxiglumide but in the area postrema was reduced by dexloxiglumide and abolished by itriglumide. We conclude that vagal afferents transmit physiological stimuli (gastrin) and pathological events (backdiffusion of luminal HCl or NH(4)OH) from the stomach to the brain stem. These communication modalities interact because, firstly, acid secretion enhances afferent signaling of gastric acid backdiffusion and, secondly, gastrin activates NTS neurons through stimulation of CCK(1) receptors on vagal afferents and of CCK(2) receptors on area postrema neurons projecting to the NTS.

We measured the energy turnover (oxygen consumption) of honeybees (Apis mellifera carnica), which were free to move within Warburg vessels. Oxygen consumption of active bees varied widely depending on ambient temperature and level of activity, but did not differ between foragers (>18 d) and middle-aged hive bees (7-10 d). In highly active bees, which were in an endothermic state ready for flight, it decreased almost linearly, from a maximum of 131.4 microl O(2) min(-1) at 15 degrees C ambient temperature to 81.1 microl min(-1) at 25 degrees C, and reached a minimum of 29.9 microl min(-1) at 40 degrees C. In bees with low activity, it decreased from 89.3 microl O(2) min(-1) at 15 degrees C to 47.9 microl min(-1) at 25 degrees C and 14.7 microl min(-1) at 40 degrees C. Thermographic measurements of body temperature showed that with increasing activity, the bees invested more energy to regulate the thorax temperature at increasingly higher levels (38.8-41.2 degrees C in highly active bees) and were more accurate. Resting metabolism was determined in young bees of 1-7 h age, which are not yet capable of endothermic heat production with their flight muscles. Their energy turnover increased from 0.21 microl O(2) min(-1) at 10 degrees C to 0.38 microl min(-1) at 15 degrees C, 1.12 microl min(-1) at 25 degrees C, and 3.03 microl min(-1) at 40 degrees C. At 15, 25 and 40 degrees C, this was 343, 73 and 10 times below the values of the highly active bees, respectively. The Q(10) value of the resting bees, however, was not constant but varied in a U-shaped manner with ambient temperature. It decreased from 4.24 in the temperature range 11-21 degrees C to 1.35 in the range 21-31 degrees C, and increased again to 2.49 in the range 30-40 degrees C. We conclude that attempts to describe the temperature dependence of the resting metabolism of honeybees by Q(10) values can lead to considerable errors if the measurements are performed at only two temperatures. An acceptable approximation can be derived by calculation of an interpolated Q(10) according to the exponential function (V(O(2))=0.151 x 1.0784(T(a))) (interpolated Q(10)=2.12).

The new optical device, Lipometer, permits the noninvasive, quick, safe, and precise measurement of the thickness of subcutaneous adipose tissue (SAT) layers at any given site of the human body. Fifteen anatomically well-defined body sites from neck to calf describe the SAT topography (SAT-Top) like an individual "fingerprint." SAT-Top was examined in 33 women with polycystic ovary syndrome (PCOS), in 87 age-matched healthy controls and in 20 Type-II diabetic women. SAT-Top differences of these three groups were described, and, based on a hierarchical cluster analysis, two distinctly different groups of PCOS women, a lean (PCOS(L)) and an obese (PCOS(O)) cluster, were found. For visual comparison of the different types of body fat distribution, the 15-dimensional body fat information was condensed to a two-dimensional factor plot by factor analysis. For comparison of the PCOS like body fat distribution with the "healthy" fat pattern, the (previously published) SAT-Top results of 590 healthy women and men (20-70 years old) and 162 healthy girls and boys (7-11 years old) were added to the factor plot. PCOS(O) women showed a SAT-Top pattern very similar to that of women with Type-II diabetes, even though the diabetic women were on average 30 years older. Compared with their healthy controls, SAT-Top of these PCOS(O) patients was strongly skewed into the android direction, providing significantly decreased leg SAT development and significantly higher upper body obesity. Compared with healthy women, PCOS(L) patients had significantly lower total SAT development (even though height, weight, and body mass index did not deviate significantly), showing a slightly lowered amount of body fat in the upper region and a highly significant leg SAT reduction. This type of fat pattern is the same as found in girls and boys before developing their sex specific body fat distribution. We conclude that women with PCOS develop an android SAT-Top, but compared in more detail, we found two typical types of body fat distribution: the "childlike" SAT pattern in lean PCOS patients, and the "diabetic" body fat distribution in obese PCOS women.

In order to survive cold northern winters, honeybees crowd tightly together in a winter cluster. Present models of winter cluster thermoregulation consider the insulation by the tightly packed mantle bees as the decisive factor for survival at low temperatures, mostly ignoring the possibility of endothermic heat production. We provide here direct evidence of endothermic heat production by 'shivering' thermogenesis. The abundance of endothermic bees is highest in the core and decreases towards the surface. This shows that core bees play an active role in thermal control of winter clusters. We conclude that regulation of both the insulation by the mantle bees and endothermic heat production by the inner bees is necessary to achieve thermal stability in a winter cluster.

In queen honey bees the free amino acid content in the haemolymph clearly depends on the physiological function and social environment of the individual. While in drones and workers the content of free amino acids increases after emergence until it reaches a peak in 5-day-old animals and decreases afterwards, the amino acid content in queens reaches its highest level (>60 nmol/ microl haemolymph) with the onset of egg laying (10 d of age). This level is about 2.5 times more than the highest level found in workers. Queens maintain this high level also when they are older (>30 d) and continue to lay eggs in average colonies. As in drones and workers, in queens the predominant amino acid is proline, which accounts for more than 50% of the total content of free amino acids in egg-laying individuals. When 10-day-old queens are prevented from mating and do not lay eggs, their amino acid content is significantly lower compared to laying queens of the same age. Also the social environment influences the contents of free amino acids in queens. When virgin queens were kept for 6 days with 20 worker bees and sufficient honey and pollen in an incubator, they had significantly lower concentrations of amino acids than virgin queens living for the same period with about 8000 workers in a colony. Most probably, the high amino acid concentration in the haemolymph is the basis for the high protein synthesis activity of laying queens.

We observed the impact of bad pollen supply (non-foraging due to artificial rain and pollen removal under poor-foraging conditions) on the survival of honey bee larvae, and on the total development time from egg-laying to the capping of a larval cell. Five days of non-foraging led to cannibalism of larvae younger than 3 days old and to a shortening of the time until larvae were sealed, but 4- and 5-day-old larvae survived even worse pollen supply situations. Manual pollen removal and reduction of income (pollen trap) induced cannibalism of younger larvae. The larvae's mean capping age significantly correlated with the mean pollen income: the less pollen was stored by the hive during the larvae's development, the earlier the larvae were capped. Both behavioral patterns lead to a quick reduction in the amount of unsealed older brood in response to a shortage of available protein. Older larvae have the highest pollen demand, so this strategy compensates for a shortage of supply by reducing demand. Additionally worker jelly gets enriched by protein gained from cannibalism, and the early capping of older larvae saves the oldest part of the brood, which represents the highest broodcare investment.

American foulbrood is a widespread disease of honeybee larvae caused by the spore-forming bacterium Paenibacillus larvae subsp. larvae. Spores represent the infectious stage; when ingested by a larva they germinate in the midgut. The rod-shaped vegetative forms penetrate the larva's intestinal tissue and start multiplying rapidly, which finally kills the larva. Spores fed to adult honeybees, however, do not harm the bees. We investigated this phenomenon. Specifically, we studied the influence of the adult honeybee midgut on the vegetative growth and on the germination of spores of P. larvae larvae. We focused on two groups of adult workers that are likely to have large numbers of spores in their gastrointestinal tracts in infected colonies: middle-aged bees, which are known to remove or cannibalize dead larvae and clean brood cells, and winterbees, which do not have frequent chances to defecate. We found that midgut extract from winterbees and worker-aged bees of different colonies almost completely inhibited the growth of the vegetative stage of P. larvae larvae and suppressed the germination of spores. The inhibiting substance or substances from the adult midgut are very temperature stable: they still show about 60% of their growth-inhibiting capacity against this bacterium after 15 min at 125 degrees C. We established a method to test growth-inhibiting factors against P. larvae larvae in vitro., (Copyright 2001 Academic Press.)

Honey bees satisfy their lipid requirement by consuming pollen. The free fatty acid content of the midgut was used to quantify fat digestion. Midguts extracted from younger workers of known ages and from foragers were divided into three components: endoperitrophic region (peritrophic membrane with gut contents), extraperitrophic region and intestinal wall. Both the total amount of pollen and the amount of free fatty acids in the endoperitrophic region and in the intestinal wall depend on the bee's age. The amounts increase within the 1st 3 days of a honey bee's life, reach maxima around the age of 8 days and then decrease continuously to the lowest values, measured in forager bees. Forced feeding with triacylglycerol results in significantly higher levels of free fatty acids, especially in the endoperitrophic region, in 8-day-old bees and foragers. This indicates that lipolytic activity depends on age and that the free fatty acid content in 8-day-old bees is primarily limited by the amount and availability of lipids ingested. The results show further that fat digestion depends on the functional status of honey bees, as is the case for pollen consumption, speed of transport of pollen bolus through the alimentary canal and protein digestion.