The control of the proventriculus in the honeybee (Apis mellifera carnica L.) II. Feedback mechanisms

The mechanisms underlying the control of solution transport rates through the proventriculus in foraging honeybees were investigated in individuals trained to collect defined amounts of sugar solutions. Following feeding, bees were injected either with metabolisable (glucose, fructose, trehalose), or non-metabolisable (sorbose) sugars, in order to distinguish between haemolymph osmolarity and haemolymph sugar levels as factors controlling the solution transport rates through the proventriculus. After a fixed period, workers were dissected in order to measure crop content and haemolymph sugar titers. Between feeding and dissection, the metabolic rate of every investigated forager was measured using open-flow respirometry. Bees injected with metabolisable sugars 15 min after feeding were observed to reduce their solution transport rates through the proventriculus, but injection of non-metabolisable sugars had no influence on them. This suggests that the solution transport rate through the proventriculus is controlled by the concentration of metabolisable compounds in the haemolymph, and not by the haemolymph osmolarity. A period of 10 min after injection of metabolisable sugars was enough to observe reduced solution transport rates. However, if bees were injected only 5 min after feeding, no reduced solution transport rates were observed 10 min after injection.     

The control of the proventriculus in the honeybee (Apis mellifera carnica L.) I. A dynamic process influenced by food quality and quantity?

The control of crop emptying in foraging honeybees was investigated in individuals trained to collect defined amounts of sugar solutions. Following feeding, they were dissected after fixed periods of time in order to measure crop content and haemolymph sugar titers. Between feeding and dissection, the metabolic rate of every investigated forager was measured using open-flow respirometry, so as to assess the effects of both food quality (concentration, molarity and viscosity of the fed sugar solution) and food quantity on the transport rate through the proventriculus. The sugar transport rate through the proventriculus was observed to be mainly dependent on the metabolic expenditure of the individual. Bee foragers were able to precisely adjust the sugar transport rate to their metabolic rates, but under certain conditions, an excess of sugars was transported through the proventriculus, more than needed to cover the bee’s energetic demands. This excess depended on the nutritive value and quantity of the fed sugar solution, and on the time after feeding. It did not depend on the metabolic rate of the bee, the molarity, or the viscosity of the fed sugar solution. As long as the bees did not exhaust their crop contents, the haemolymph sugar titers were unaffected by this excess amount transported, by the time after feeding, the concentration and the viscosity of the fed sugar solution. For all feeding conditions assayed, the haemolymph trehalose titer remained constant, while the titers of other haemolymph sugars varied. It is suggested that the trehalose concentration in the haemolymph is regulated in honeybees, and that it represents the controlled variable in the feedback loop responsible for the transport rate through the proventriculus.     

Feeding glucose or sucrose, but not trehalose, suppresses the starvation-induced premature pupation in the yellow-spotted longicorn beetle, Psacothea hilaris

Under 25 degrees C and a long-day photoperiod, starvation induces premature pupation in 4th instar Psacothea hilaris larvae exceeding a threshold weight of 180 mg, resulting in the formation of small but morphologically normal adults. To investigate possible mechanisms underlying this phenomenon, we first measured the hemolymph trehalose and glucose levels of starved larvae. When larvae were starved after 4 days of feeding (attaining the threshold weight), glucose levels decreased 4-fold within the next 24 h, while trehalose levels, after a temporary slight decrease, increased remarkably to reach a peak just before the prepupa stage. The effects of ingesting various nutrients on the developmental fate and the hemolymph sugar titers of starving larvae were then examined. Feeding on agar blocks containing sucrose or glucose totally suppressed the occurrence of premature pupation, while trehalose, fructose, casein and starch were ineffective. Feeding on glucose or trehalose resulted in a 6-fold decrease in hemolymph glucose levels and remarkably elevated trehalose levels. Since feeding on glucose and trehalose induced similar changes in hemolymph sugar titers but trehalose was not effective in suppressing premature pupation, glucose may have exhibited its effects via gustatory mechanisms.     

高温对家蚕三品系血淋巴中糖水平的影响（英文）

家蚕Bombyx mori的两个二化性品系热耐受型NB4D2和热敏感型CSR2均适合于温带气候,而多化性的PM（Pure Mysore) 品系适合于热带气候,将这3种品系5龄幼虫分别置于32℃和36℃的高温下,观察高温对其5龄幼虫至蛹期血淋巴中糖含量及海藻糖酶活性的影响。结果表明： PM幼虫和蛹的死亡率均小于NB4D2和CSR2。在蜕皮期间血淋巴海藻糖水平较高,而葡萄糖水平及海藻糖酶活性较低。32℃和36℃的高温下,幼虫蜕皮期间血淋巴中糖含量及海藻糖酶活性仅在其各自的水平上表现为小幅度的增加。蜕皮后幼虫血淋巴中海藻糖含量显著下降,而葡萄糖含量和海藻糖酶活性显著上升。在较高温度下,蜕皮后幼虫血淋巴中海藻糖含量下降幅度更大,而葡萄糖含量及海藻糖酶活性上升水平也更加显著。25±1℃下取食幼虫血淋巴中葡萄糖含量显著下降,海藻糖含量显著上升;3℃和36℃下PM 和NB4D2取食幼虫血淋巴葡萄糖和海藻糖含量以及海藻糖酶活性增加,而CSR2均减少或降低。吐丝幼虫血淋巴中葡萄糖含量及海藻糖酶活性显著下降,海藻糖小幅度下降。而在较高温度下,耐热型PM 和NB4D2吐丝家蚕血淋巴糖含量含量和海藻糖酶活性明显增加,而热敏感型CSR2的则明显下降。这3种品系蛹发育期的血淋巴糖含量及海藻糖酶活性均下降。在两较高温度下,PM蛹期血淋巴糖和海藻糖酶活性增加,而NB4D2 36℃时增加幅度小于32℃时。对于CSR2,32℃时观察到其血淋巴葡萄糖含量增加,但当环境温度增加到36℃时其血淋巴葡萄糖含量降至正常水平下。然而,当CSR2的蛹置于32℃和36℃时血淋巴海藻糖含量及其酶活性下降,且36℃时下降幅度更大。因此,桑蚕对高温的适应取决于家蚕的品系及发育阶段,并可通过其血淋巴糖及海藻糖酶活性水平进行验证。     

Resorption und Stoffwechsel von Glucose im Darmtrakt der Wanderheuschrecke Locusta migratoria

Absorption and metabolic fate of ¹⁴C-glucose after injection into mid-gut preparations from Locusta migratoria were studied in vitro. After 5 min nearly half of the total radioactivity had already passed into the incubation medium and in 2 hr there was an increase to 75 per cent. The radioactivity in the medium was found mainly in glucose and fructose, which were approximately equally labelled, and a minor part appeared in the organic acid and sugar phosphate fraction. This pattern of labelling changed drastically when non-radio active glucose was present in the incubation mixture. In these experiments an extensive synthesis of sucrose could be demonstrated.

Sucrose formation from UDP-¹⁴C-glucose by mid-gut homogenates was stimulated by fructose-6-phosphate.

The appearance of radioactivity in the contents of the gut and in the faeces was demonstrated in in vivo experiments after injection of ¹⁴C-glucose into the haemolymph.

Sugar concentrations in the haemolymph of locusts were measured after 24 hr starvation and after feeding. The levels of glucose, fructose, and sucrose were very low in the starving animals and increased ten- to twenty-fold after feeding. The trehalose concentrations remained high and relatively constant.

The activities of 18 enzymes of the intermediary metabolism were assayed in the fore-, mid-, and hindgut.     

Gustatory perception and metabolic utilization of sugars by<Emphasis Type="Italic"> Myrmica rubra</Emphasis> ant workers

The suitability of various nectar and honeydew sugars as a food source for the polyphagous ant species M. rubra (L.) was studied. The sugars used included monosaccharides (fructose, glucose, galactose, mannose, rhamnose), disaccharides (sucrose, maltose, trehalose, melibiose, lactose) and trisaccharides (melizitose, raffinose, erlose). Single-sugar solutions were tested on ant workers in a long-term laboratory bioassay in which acceptance of the solutions and ant survival were recorded. The acceptance of the sugars was confirmed in a second bioassay in which feeding time was established. Enzymatic hydrolysis of sucrose, maltose and melibiose was investigated through HPLC analyses of workers fed these disaccharides. Sugar acceptance and feeding time were related to ant survival. Considering the monosaccharide units of which the sugars are composed, fructose seems especially suitable as a short-term energy source, while glucose appears to be used both directly and for storage. The presence of a galactose unit appears to reduce sugar suitability. It is suggested that the workers possess invertase and maltase and to a lesser degree also galactosidase. The gustatory perception is correlated with the profitability of sugars in further metabolic processes.     

Aphid–ant mutualism: how honeydew sugars influence the behaviour of ant scouts

Honeydew is the keystone on which ant–aphid mutualism is built. The present study investigates how each sugar identified in Aphis fabae Scopoli honeydew acts upon the feeding and the laying of a recruitment trail by scouts of the aphid‐tending ant Lasius niger Linnaeus, and thus may enhance collective exploitation by the ant mutualists. The feeding preferences shown by L. niger for honeydew sugars are: melezitose = sucrose = raffinose > glucose = fructose > maltose = trehalose = melibiose = xylose. Although feeding is a prerequisite to the launching of trail recruitment, the reverse is not necessarily true: not all ingested sugar solutions elicit a trail‐laying behaviour among fed scouts. Trail mark laying is only triggered by raffinose, sucrose or melezitose, with the latter sugar being specific to honeydew. By comparing gustatory and recruitment responses of ant foragers to sugar food sources, the present study clarifies the role of honeydew composition both as a source of energy and as a mediator in ant–aphid interactions. Lasius niger feeding preferences can be related to the physiological suitability of each sugar (i.e. their detection by gustatory receptors as well as their ability to be digested and converted into energy). Regarding recruitment, the aphid‐synthesized oligosaccharide (melezitose) could be used by ant scouts as a cue indicative of a long‐lasting productive resource that is worthy of collective exploitation and defence against competitors or aphid predators.     

Parasitic infection leads to decline in hemolymph sugar levels in honeybee foragers

Parasites by drawing nutrition from their hosts can exert an energetic stress on them. Honeybee foragers with their high metabolic demand due to flight are especially prone to such a stress when they are infected. We hypothesized that infection by the microsporidian gut parasite Nosema ceranae can lower the hemolymph sugar level of an individual forager and uncouple its energetic state from its normally tight correlation with the colony energetic state. We support our hypothesis by showing that free-flying foragers that are infected have lower trehalose levels than uninfected ones but the two do not differ in their trehalose levels when fed until satiation. The trehalose level of infected bees was also found to decline at a faster rate while their glucose level is maintained at a quantity comparable to uninfected bees. These results suggest that infected foragers have lower flying ability and the intriguing possibility that the carbohydrate levels of an individual bee can act as a modulator of its foraging behavior, independent of social cues such as colony demand for nectar. We discuss the importance of such pathophysiological changes on foraging behavior in the context of the recently observed colony collapses.     

The parasitic mite <Emphasis Type="Italic">Varroa destructor</Emphasis> affects non-associative learning in honey bee foragers, <Emphasis Type="Italic">Apis mellifera</Emphasis> L.

The parasitic mite Varroa destructor influences flight behavior, orientation and returning success of forager honeybees (Apis mellifera) infested as adults. As impaired orientation toward the nest entrance might be due to deficiency in recognition and responsiveness to stimuli in the environment, we examined effects of V. destructor on sensory responsiveness, non-associative and associative learning of honey bee foragers by using proboscis extension reaction paradigm (PER). Although infested and uninfested workers were initially equally responsive to different concentrations of sugar water, we found differences in non-associative learning. In habituation, PER to repeated sugar stimulation of the antennae occurred faster in infested foragers compared to uninfested foragers. In sensitization, infested foragers showed a lower response to an odor stimulus following sugar stimulation than non-infested foragers. Differences in non-associative paradigms were more pronounced in bees with lower responsiveness to sucrose. In conditioning learning experiments, a significant reduction in proboscis extension response was found 1 min but not 12 min after a single conditioning trial indicating that V. destructor predominantly affects the non-associative components of learning and its underlying neural and molecular processes. Jasna Kralj and Axel Brockmann have contributed equally to this study.     

Sugar convertibility in the parasitoid Cotesia glomerata (Hymenoptera: Braconidae)

Lack of suitable sugar sources for adult parasitic wasps is an important cause of failure in biological control programs, but the metabolic constraints of sugar feeding are poorly understood. Here we investigated the suitability of 11 naturally occurring sugars as energy sources for the parasitoid Cotesia glomerata (L.) (Hymenoptera: Braconidae). By feeding energy-deprived individuals with given quantities of a 40% (w:w) sugar solution, we assessed recovery time and lifespan after feeding. More than 50% of the wasps recovered within 20 min, and at least 80% within 60 min after uptake of one of the monosaccharides fructose or glucose, the disaccharide sucrose, or the trisaccharide melezitose. Between 40 to 80% of the test insects recovered within an hour after intake of maltose, raffinose, galactose, or mannose. Less than 25% recovered within 1 h after uptake of melibiose, trehalose, rhamnose, or water (control). Parasitoids obtained the highest lifespan benefits after intake of glucose, fructose, sucrose, or melezitose, indicating and confirming their convertibility as an energy source for C. glomerata. In contrast, no lifespan increase was found after consumption of rhamnose and trehalose. The differences in recovery time and lifespan are discussed in terms of parasitoid enzyme activity and metabolic processes.     

<Emphasis Type="Italic">Vespula germanica</Emphasis> Foragers Do Not Scent-Mark Carbohydrate Food Sites

We tested whether foragers of Vespula germanica leave a chemical attractant at a carbohydrate food site. Foragers were allowed to make 50 or 100 visits to a feeder filled with a rich, unscented fructose solution. We then gave them a choice between this and an identical but unvisited feeder. There was no difference between the number of individuals that chose the visited feeder and those that chose the unvisited one. Instead, wasps showed a tendency to choose a feeder based on its relative position on the feeding stand. We conclude that foragers of V. germanica do not scent-mark food sites. Instead, in the absence of food odor, they rely on local visual cues to relocate carbohydrate food sources.     

Feeding Behavior and Survival of Glucose-Averse Blattella germanica (Orthoptera: Blattoidea: Blattellidae) Provided Glucose as a Sole Food Source

Glucose-averse (Glu/Glu) male German cockroaches, Blattella germanica, rejected brief exposures to glucose solutions despite food deprivation so extreme as to cause high mortality. In the 24-h period following 5 days of food deprivation, both Glu/Glu and wild-type (+/+) were given a continuous source of 1 M glucose solution. Although Glu/Glu visited the solution as frequently as +/+, feeding bouts were significantly shorter. These truncated feeding bouts were sufficient to extend Glu/Glu longevity to ca. 80 days, regardless of glucose concentration. When provided continuously, the glucose was completely rejected by Glu/Glu for the first 3 days. However, by day 9, glucose intake levels were similar to those of wild-type cockroaches. The initial feeding lag probably accounted for the lower long-term survival of the Glu/Glu vs the +/+ strain. Under the stress of food deprivation, glucose-averse B. germanica modify their feeding behavior toward glucose like many phytophagous insects provided non-host plants in no-choice situations.     

Differential expression of a fructose receptor gene in honey bee workers according to age and behavioral role

Honey bee (Apis mellifera) workers contribute to the maintenance of colonies in various ways. The primary functions of workers are divided into two types depending on age: young workers (nurses) primarily engage in such behaviors as cleaning and food handling within the hive, whereas older workers (foragers) acquire floral nutrients beyond the colony. Concomitant with this age‐dependent change in activity, physiological changes occur in the tissues and organs of workers. Nurses supply younger larvae with honey containing high levels of glucose and supply older larvae with honey containing high levels of fructose. Given that nurses must determine both the concentration and type of sugar used in honey, gustatory receptors (Gr) expressed in the chemosensory organs likely play a role in distinguishing between sugars. Glucose is recognized by Gr1 in honey bees (AmGr1); however, it remains unclear which Gr are responsible for fructose recognition. This study aimed to identify fructose receptors in honey bees and reported that AmGr3, when transiently expressed in Xenopus oocytes, responded only to fructose, and to no other sugars. We analyzed expression levels of AmGr3 to identify which tissues and organs of workers are involved in fructose recognition and determined that expression of AmGr3 was particularly high in the antennae and legs of nurses. Our results suggest that nurses use their antennae and legs to recognize fructose, and that AmGr3 functions as an accurate nutrient sensor used to maintain food quality in honey bee hives.     

Spatial organization in ants’ nests: does starvation modify the aggregative behaviour of Lasius niger species?

Ant colonies that undergo long starvation periods have to tune their exploratory and foraging responses to face their food needs. Although the number of foragers is known to increase with food deprivation in the ant Lasius niger, such enhanced food exploitation is not related to a more intense recruitment by successful scouts. We thus suggest that the colony’s response to a food shortage could result from changes at the level of the ant recruits, in particular from changes in their spatial organization inside the nest. Since aggregation plays a key role in the social organization of ants, we assume that the colony’s response to starvation could be due to changes in the aggregative behaviour of L. niger nestmates.We thus compared the aggregation dynamics of inner-nest workers and foragers having undergone either a short or a long-lasting starvation period. Whatever the ethological group (foragers or inner-nest workers), there was no significant influence of starvation on the aggregation dynamics nor on any feature of the observed clusters. This result shows that an increased foraging response to food shortage cannot be explained by changes in the tendency of nestmates to aggregate within the nest. Finally, we discuss other behavioural mechanisms, in particular changes in behavioural thresholds that could underlie the adaptive changes seen in colony foraging after long starvation periods. Received 25 June 2007; revised 21 January 2008; accepted 24 January 2008.     

Effects of age and blood sugar levels on the proboscis extension of the blow fly Phormia regina

In some insects the proboscis is extended to imbibe a sugar solution if the concentration of sugar applied to the chemosensilla exceeds the behavioural threshold value. Recently, I found a reversal of the threshold values of this "proboscis extension reflex" (PER) in the blow fly (Phormia regina M.) for glucose and fructose. It depended on maturation and physiological conditions, both of which are explicable in terms of changing concentration of haemolymph trehalose. The direct injection of trehalose into the fly haemocoele brought about a dramatic shift of the threshold values of PER measured on tarsi or labellar sensilla, suggesting a strong dependence of PER on the blood sugar level. Using the tip-recording method, the dose-response (impulse frequency) curves for glucose and fructose were obtained on individual largest labellar chemosensilla. The curves for glucose and fructose crossed at one point because the former had a steeper gradient and higher maximum response than the latter. Injection experiments with trehalose were also carried out to test for changes in gustatory response. The shifting of the behavioural dose-response curves for glucose and fructose two hours after injection of 1 M trehalose (2 μl) into the haemocoele of the fly was associated with significant reduction in responsiveness of labellar chemosensilla to glucose, but less so to fructose. No change in responsiveness was found following injection of mannose. A hypothesis to explain the reversal relation of the PER thresholds, based on a shift in the firing rate in gustatory sensilla and possibly also interneurons, is discussed.     

Thorax vibrations of a stingless bee (<Emphasis Type="Italic">Melipona seminigra</Emphasis>). II. Dependence on sugar concentration

Using a laser vibrometer we studied the influence of the foods sugar concentration on different parameters of the thorax vibrations produced by foragers of Melipona seminigra during trophallaxis in the nest. The concentrations tested (20–70% sugar w/w) were within the biologically relevant range. They substantially influenced different parameters of the thorax vibrations. An increase of energy gains at the food source due to an increased sugar concentration was followed by an increase of both the pulse duration and the duty cycle and by a decrease of the pause duration between two subsequent pulses. These findings further support the hypothesis that the temporal pattern of the thorax vibrations reflects the energy budget of a foraging trip rather than food source distance. Likewise, the steep increase of pulse duration variability with sugar concentration is hard to reconcile with the assumption that pulse duration conveys reliable information about food source distance when bees collect at high-quality food sources.     

Nectar distribution and its relation to food quality in honeybee (<Emphasis Type="Italic">Apis mellifera</Emphasis>) colonies

In honeybees (Apis mellifera), the process of nectar collection is considered a straightforward example of task partitioning with two subtasks or two intersecting cycles of activity: (1) foraging and (2) storing of nectar, linked via its transfer between foragers and food processors. Many observations suggest, however, that nectar collection and processing in honeybees is a complex process, involving workers of other sub-castes and depending on variables such as resource profitability or the amount of stored honey. It has been observed that food processor bees often distribute food to other hive bees after receiving it from incoming foragers, instead of storing it immediately in honey cells. While there is little information about the sub-caste affiliation and the behaviour of these second-order receivers, this stage may be important for the rapid distribution of nutrients and related information. To investigate the identity of these second-order receivers, we quantified behaviours following nectar transfer and compared these behaviours with the behaviour of average worker hive-bees. Furthermore, we tested whether food quality (sugar concentration) affects the behaviour of the second-order receivers. Of all identified second-order receivers, 59.3% performed nurse duties, 18.5% performed food-processor duties and 22.2% performed forager duties. After food intake, these bees were more active, had more trophallaxes (especially offering contacts) compared to average workers and they were found mainly in the brood area, independent of food quality. Our results show that the liquid food can be distributed rapidly among many bees of the three main worker sub-castes, without being stored in honey cells first. Furthermore, the results suggest that the rapid distribution of food partly depends on the high activity of second-order receivers. Received 31 August 2006; revised 8 December 2006; accepted 11 December 2006.     

Glucose and fructose consumption in a phosphoglucoseisomeraseless mutant in Saccharomyces cerevisiae

Saccharomyces cerevisiae with a practically complete absence of phosphoglucoseisomerase activity when grown in fructose or glucose minimal medium showed different consumption of fructose and glucose during different periods of the culture. At the beginning of growth, cells had a great quantity of glucose available relative to their requirements and a large quantity of trehalose accumulated from ¹⁴C-glucose in comparison with the wild type strain. A second phase arises when the concentration of glucose in the medium was practically absent and the cells obtain glucose by mobilisation of stored glucose containing compounds. It is very likely that at this moment a balance rate between glucose 6-phosphate formation and consumption occurs. Finally cells reach conditions of glucose starvation and fructose consumption increases in this last stage. The different consumption of fructose throughout different periods of cell growth most probably indicates a strict regulation at the level of sugar uptake.Non Standard Abbreviation pgi phosphoglucoseisomerase     

Production of carotenoids by β-ionone-resistant mutant of <Emphasis Type="Italic">Xanthophyllomyces dendrorhous</Emphasis> using various carbon sources

Batch culture kinetics of the red yeast, Xanthophyllomyces dendrorhous SKKU 0107, revealed reduction in biomass with glucose and lower intracellular carotenoid content with fructose. Figures were different when compared to sucrose, which is a disaccharide of glucose and fructose. In contrast, specific growth rate constant stayed between 0.094~0.098 h⁻¹, irrespective of the carbon sources employed. Although the uptake rate of glucose was found to be 2.9-fold faster than that of fructose, sucrose was found to be a more suitable carbon source for the production of carotenoids by the studied strain. When sugar cane molasses was used, both the specific growth rate constant and the intracellular carotenoid content decreased by 27 and 17%, respectively. Compared with the batch culture using 28 g/L sugar cane molasses, fed-batch culture with the same strain resulted in a 1.45-fold higher cell yield together with a similar level of carotenoid content in X. dendrorhous SKKU 0107.     

Hexitols as major intermediates of glucose assimilation by mycelium of Puccinia graminis

Mycelium of Puccinia graminis was grown for 4 d on 200 mM D-[U-¹⁴C]glucose followed by a cold chase for 30 h. Analysis of cellular metabolites during the chase indicated significant turnover only in carbohydrates soluble in 80% (w/v) ethanol. A kinetic analysis of the depletion of [¹⁴C] in pools of free sugars and sugar alcohols indicated that the trehalose pools and a small proportion (12–16%) of the mannitol and glucitol pools did not turn over, whilst pools of glucose, fructose, and the remainder of the hexitols became totally,depleted of label during the chase. Because the [¹⁴C] was totally lost from the pools of glucose and fructose prior to the hexitols, it was deduced that both of these hexoses were precursors of the hexitols. Estimation of the carbon fluxes through pools indicated that 52, 36 and 16% of the carbon from glucose was assimilated via glucitol, fructose and mannitol respectively, demonstrating that glucitol could not have originated from fructose as sole precursor. After offering D-[U-¹⁴C]glucitol, [¹⁴C] was assimilated into trehalose phosphate, glucans and amino acids, but not into free glucose or fructose. These data indicate that hexitols are quantitatively important intermediates during the assimilation of glucose by Puccinia graminis.