Abdominal distention terminates subsequent host-seeking behaviour of Aedes aegypti following a blood meal

The failure of Aedes aegypti females to engage in host-seeking behaviour following a replete blood meal was investigated. Abdominal distention appears to be responsible for this immediate inhibition after feeding. Large enemas of saline had the same effect as blood in terminating host-seeking; this was not due merely to the presence of large amounts of fluid, but rather to the distention produced by these liquids. Since transection of the ventral nerve cord anterior to the 2nd abdominal ganglion did not release the inhibition in blood-fed females, we restricted the degree of distention of abdominal segments with wax. Distention of the abdomen anteriorly by a blood meal more effectively inhibited host-seeking than did distention posteriorly, suggesting that stretch receptors in the anterior portion of the abdomen regulate the response towards a host.     

Temporary feeding inhibition caused by artificial abdominal distension in the bedbug, Cimex lectularius

Abdominal distension of haematophagous insects caused by ingested blood has been recognised as an important contributor to triggering meal termination, feeding inhibition and further susceptibility to host signals. Factors that regulate feeding behaviour of the common bedbug, Cimex lectularius, are poorly understood. By injecting air directly into the body cavity of virgin female C. lectularius we artificially induced abdominal distension without providing chemical cues of the blood meal and without applying gut distension. Body length increased to 138% after feeding and 147% after inflation. The early decline in body volume is similar in blood-fed bugs but after between 8 and 24h became faster in inflated than fed bedbugs. Artificially inflated individuals remained feeding-inhibited at lower abdominal distensions than those that terminate blood ingestion (to ca. 135% initial body length, or up to about 5h). Feeding activity resumed earlier in inflated than blood-fed bugs. These results suggest that artificial abdominal distension has an inhibitory effect on feeding but is not the sole mechanism in preventing further feeding.     

Reproductive physiology of Anopheles gambiae and Anopheles atroparvus.

When exposed to a human host, Anopheles gambiae started probing 4 h post-eclosion, but 95% successfully blood-fed by 16-20 h with maximal blood volumes of 5- 10 microl per female. When fed sugar, the 95% feeding was not observed until 36-40 h post-eclosion; sugar meals appeared to interfere with blood meals. Similarly in An. atroparvus, maximum volumes were 10 microl when starved but only 6 microl when fed sugar. This species did not bite before 2 d, and 95% biting was by 4 d. Given single blood meals to water-kept An. gambiae, a threshold body size for oogenesis was detected. With wing lengths below 2.8 mm, eggs never matured, but when sugar-fed, females of all sizes matured eggs including the synthesis of maternal deposits. Although sugar feeding interfered with blood feeding, more lipid was transferred to the yolk. In water-kept An. atroparvus only 5% of the females produced eggs. When sugar-fed for 4 d, all females matured eggs, so in this species sugar feeding appeared to be essential for oogenesis. An. gambiae always took multiple blood meals, tested at any time after the first ones, leading to 120 mature eggs/female. Yolk composition was 3.9 mcal protein and 3.8 mcal lipid/oocyte when kept on water, but 2.8 meal protein and 4.3 mcal lipid/oocyte with intermittent sugar meals, thus marking a surprising flexibility in synthesis of yolk protein and lipid that strongly depends on additional carbohydrates sources. Only 80% of water-fed An. atroparvus re-fed 2 d after a first blood meal with small females taking three blood meals but they still showed reduced fecundity. Only the large water-fed females matured eggs, with blood volumes higher than 9-12 microl. When fed sugar, the blood meal input was reduced, but oogenesis was possible, whereas water-fed females required three blood meals to reach the caloric level comparable to pre-feeding sugar-fed females. Water-fedAn. gambiae could survive on daily blood meals alone, but survival was further extended by intermittent sugar meals. When offered a blood donor daily, there was a behavioral difference. Females maintained alone showed a more or less regular 3 d feeding and oviposition activity, while females kept in groups fed daily followed a daily oviposition pattern, suggesting gonotrophic discordance.     

The effect of deprivation on the microstructure of feeding by the Tobacco hornworm caterpillar

An automated device was used to examine, in detail, feeding on disks of wheat germ medium by fifth-instar Manduca sextacaterpillars. Comparisons were made between some animals which had ad libitum access to food at all times and others which were deprived of food for 1–5 h before being tested. Feeding patterns of both groups indicated regulation of feeding both between and within meals. Deprived animals ate more during their first meal than did nondeprived animals chiefly by increasing (a) the number of chewing bouts (and thus the meal duration) and (b) the bite frequency. Calculations indicated that the deficit caused by deprivation was made up during the first meal. However, deprived animals continued to eat more than nondeprived ones in subsequent feeding also. Passage of food through the gut was examined by dissecting out the contents of each region of the gut at various times after a colored test meal. Food passed through the foregut directly into the anterior part of the midgut. It stayed in the middle third of the midgut longer than in the anterior and posterior thirds, and the first pellet resulting from the test meal appeared 4 h after the meal. The following mechanisms of feeding regulation are proposed: (a) volumetric feedback mediated by stretch receptors of the foregut and anterior third of the midgut which terminates meals; (b) the development and subsequent reduction of satietyspecific behaviors mediated either by stretch receptors or by some other means which, e.g., allow the next meal to begin; and (c) metabolites whose levels drop during deprivation, triggering a series of events which lead to the excess feeding observed.     

The control of meal termination in the locust

The relationship between the amount eaten during a meal, meal duration and ingestion rate was investigated in the locust, Locusta migratoria (L.). The results suggested a relatively simple model for the control of meal termination whereby a variety of exogenous and endogenous factors, including chemosensory stimuli provided by the food, the presence of other locusts nearby, age and deprivation-related factors, are integrated within the central nervous system and together contribute to the level of excitation present at the start of a meal. The amount of excitation determines both the mean rate of ingestion during the meal and the amount of inhibition needed to end the meal. Since much of the inhibitory feedback comes from gut stretch, meal size and mean ingestion rate are positively correlated, in fact increasing proportionally, and, hence, meal duration remains constant. At high levels of excitation, however, ingestion becomes rate limited and meal duration subsequently increases proportionally with meal size. Changes in the rate of ingestion throughout a meal were also investigated. The rate declined exponentially but, unlike in the rat, the initial ingestion rate remained constant with changes in excitation and the decay function varied such that small meals began at the same ingestion rate as large meals but the rate of decline was subsequently faster.     

Regulation of sensitivity in the peripheral chemoreceptor systems for host-seeking behaviour by a haemolymph-borne factor in Aedes aegypti

The taking of a blood meal and subsequent development of eggs by a female mosquito is reported to suppress host-seeking behaviour. This change in behaviour may be partly mediated by changes in certain chemosensitive antennal afferent neurones that influence the behaviour of female mosquitoes. Electrophysiological activity of the lactic acid-excited neurons to lactic acid—a normal host-attractant substance—is depressed following a blood meal. This reduction in lactic acid sensitivity is coincident with the reported inhibition of host-seeking behaviour. The reduction in lactic acid sensitivity is reversible; this sensitivity returns to the pre-blood-fed level following oviposition. Like the inhibition of host-seeking behaviour, the reduced lactic acid sensitivity is due to a transfusable, haemolymph-borne factor. A role for the peripheral sensory system in the control of behaviour in female mosquitoes is discussed.     

COURTSHIP FEEDING AND THE FITNESS OF FEMALE KATYDIDS (ORTHOPTERA: TETTIGONIIDAE)

Katydid males (Requena verticalis) produce spermatophores with a large sperm-free spermatophylax, which is eaten by the female after mating. In this study, I asked 1) how do spermatophylax nutrients affect the fitness of the mated female and her progeny? 2) does this male-produced food substitute for other food in the diet of the female, or is it a source of specialized nutrients? and 3) does an increase in the size of the spermatophylax eaten influence female reproduction in the same way as the additional spermatophylaxes that would be obtained from multiple mating? An experimental increase either in dietary protein or the number of spermatophylaxes eaten increased the number of eggs produced. However, spermatophylax size had no effect on egg number. An increase in either the size or number of spermatophylaxes eaten resulted in an increase in egg size. There was no influence of protein in the general diet on egg size. This suggests that males provide nutrition not available from other sources. Although there were no direct effects of number of spermatophylaxes eaten by females on the overwintering survival of their progeny, offspring from females producing larger eggs had a relatively higher probability of surviving winter. The amount of spermatophylax eaten had no influence on the mean adult size of progeny but significantly increased the mean date at which sons matured to adulthood. There was no influence of dietary protein on these variables. Since maturation date is positively correlated with adult size in both sexes, it is suggested that the influence of courtship feeding on maturation date may result in an increase in adult size and thus the fitness of sons. A significant correlation between the size of the female and the mean size of her sons (but not daughters) suggests that there is also a heritable component to body size.     

Courtship role reversal and deceptive signals in the long-tailed dance fly, Rhamphomyia longicauda

We examined the function of secondary sexual characters in the role-reversed, lekking behaviour of female long-tailed dance flies, Rhamphomyia longicauda Loew (Empididae), to test the hypothesis that the degree of abdominal distention is an honest female signal about the state of egg development. Female Rhamphomyia cannot hunt for prey and they receive all of their protein from males by exchanging copulations for nuptial prey gifts. Females compete for male gifts within leks that are organized for a brief period each evening before dark. Before hovering within leks, females swallow air, inflating expandable pouches on the pleural margins of the abdomen. The result is a large saucer-like abdomen which is further exaggerated by wrapping scaled pro-, meso- and metathoracic legs along its pleural margins. Male preference for an enlarged abdomen was confirmed by suspending plastic models of varying size from monofilament lines and recording which models attracted the most males. There was a positive relationship between egg development and abdominal distention in a related species, R. sociabilis (Williston), which lacks inflatable abdominal pouches. Multiple regression showed that in R. longicauda, abdominal inflation completely masks the state of egg development. We conclude that female R. longicauda deceive mate-seeking males with the unreliable message that eggs are nearing maturation in order to obtain a protein meal in exchange for copulation. Males that fail to identify a female bearing mature eggs risk near-certain cuckoldry and an increased probability that the female will die before oviposition. Copyright 2000 The Association for the Study of Animal Behaviour.     

The role of hemolymph proline as a nitrogen sink during blood meal digestion by the mosquito Aedes aegypti

Mosquitoes utilize the amino acids derived from blood meal protein to produce egg proteins. But the amino acids can also be used to produce egg lipid or can be oxidized for energy production. These latter two processes result in the release of nitrogen as toxic ammonia. Therefore, amino acids must be processed in such a way that amino acid nitrogen can be incorporated into non-toxic waste products. Proline is the predominant amino acid in the hemolymph of the adult female mosquito Aedes aegypti. After feeding on albumin meal, hemolymph proline levels increased five-fold over unfed levels, reached maximal levels in the first hours after feeding and remained high through oviposition. Hemolymph proline levels increased as the concentration of protein in the meal increased. When starved of sugar for 24 h prior to feeding on an albumin meal, hemolymph proline levels increased four-fold over the proline levels of non-starved mosquitoes. Proline levels after feeding on a protein deficient in essential amino acids, pike parvalbumin, increased to twice the levels of albumin fed mosquitoes. Based on these observations, we propose that mosquitoes utilize proline as a temporary nitrogen sink to store ammonia arising from deamination of blood meal amino acid.     

Arrest, resorption, or maturation of oöcytes in Aedes aegypti: dependence on the quantity of blood and the interval between blood meals

ABSTRACT. After emergence, the follicles of A. aegypti double in length and the oöcytes may deposit a small amount of yolk, but within 2 days growth is arrested. Renewed growth and vitellogenesis, as well as the number of eggs finally produced, depends on the quantity of blood ingested. All females, given either a small (1 μl) or large (4 μl) meal of rat blood by enema, began yolk deposition in a nearly equal number of oöcytes, and each oöcyte had about the same amount of yolk 8 h later. Within 48 h, females fed 4 μl had each produced more than 100 eggs, whereas females fed 1 μl either had continued yolk deposition in some oöcytes, while most degenerated, or had all re-entered oögenic arrest. Consequently, 48 h after the 1 μl meal, a female had either c. 50 or 0 eggs. Even by 14 h after a 1-μl meal, females were either committed to re-enter oögenic arrest or to complete maturation of some oöcytes and resorb the yolk of others. This was shown by removing and examining one ovary 14 h after a blood meal and then giving a second blood meal. The second meal stimulated meal maturation in the remaining ovary, but only in those females whose oöcytes had been in oögenic arrest when the first ovary was examined; the second meal had no effect on females whose first ovary had contained both vitellogenic and degenerating oöcytes. Oösorption was not reversed by a second blood meal. Our results do not support the hypothesis that the female 'evaluates' the ingested meal and begins vitellogenesis in an 'appropriate' number of oöcytes. The results demonstrate that the ovary is an unreliable indicator of the frequency of blood-feeding, when females take a small meal.     

Delayed allatectomy and feeding in Phormia regina (Meig): effects on follicular development.

A delay in feeding a protein meal to the female blow-fly following allatectomy results in inhibition of follicular development in the previtellogenic stage. On the other hand, a protein meal given immediately after the operation permits 55% of the flies that do not deposit yolk in their oocytes fail to do so because of the absence of a corpus allatum hormone necessary for the synthesis of vitellogenin.     

Recurrent nerve inhibition of protein feeding in the blowfly Phormia regina

ABSTRACT. Stretch receptors located in the foregut appear to contribute negative feedback to the ingestion of protein by Phormia ; as fluid is consumed, the concomitant stretch impedes further drinking. This inhibitory feedback, transmitted to the frontal ganglion via the recurrent nerve, seems to depress protein ingestion less than carbohydrate ingestion. Such a disparity may help to ensure that the female fly acquires enough protein for egg development through the retention of her ability to consume available protein even after carbohydrate consumption has been terminated.     

Metabolic fate of [14C]-labeled meal protein amino acids in Aedes aegypti mosquitoes

We developed a method to follow the metabolic fate of [(14)C]-labeled Euglena gracilis protein amino acids in Aedes aegypti mosquitoes under three different adult nutritional regimes. Quantitative analysis of blood meal protein amino acid metabolism showed that most of the carbon of the amino acids was either oxidized to CO(2) or excreted as waste. Under the three different adult nutritional regimes, no significant differences in the metabolism of amino acids were found, which indicated that the female A. aegypti mosquitoes possess a substantial capacity of maintaining metabolic homeostasis during a gonotrophic cycle. The amount of maternal glycogen and lipid after egg laying were significantly lower in the mosquitoes that underwent a partial starvation before a blood meal and/or starvation after the blood meal. The content of egg lipid or protein or the number of eggs laid did not show a significant difference among the three different regimes, which indicates that stable fecundity of A. aegypti under the partial starvation before a blood meal and/or starvation after the blood meal seemed to result from a trade-off between current fecundity and future survival after the eggs laid. The methods described in this paper can be applied to a wide range of questions about the effects of environmental conditions on the utilization of blood meal amino acids.     

Utilization of pre-existing energy stores of female Aedes aegypti mosquitoes during the first gonotrophic cycle

Pre-existing energy reserves may play an important role in regulating the utilization of blood meal proteins in female anautogenous mosquitoes. Determining the fate of reserves derived from the sugar meal and larval food during the first gonotrophic cycle would help to elucidate the relative contributions of larval and adult nutrition to survival and reproduction. We measured the allocation of pre-blood-meal reserves to egg production or energy production during the first gonotrophic cycle by using [14C]-labeled female Aedes aegypti mosquitoes. Feeding adults [3,4-14C]-glucose labeled the glycogen and sugar stores (approximately 50%), lipid stores (approximately 25%), and protein and amino acid stores (approximately 25%). During the first gonotrophic cycle, about 60% of the glycogen and sugar stores were metabolized and all were used for energy production. About 33% of the labeled protein and 72% of the labeled amino acid stores were metabolized, with about 9% being transferred to the eggs and the rest oxidized. About 30% of the lipid was metabolized, with about 65% being transferred to the eggs and the rest oxidized. Feeding [1-14C]-oleic acid to larvae effectively labeled adult lipid stores with about 75% of the label in lipid stores and 16% in proteins and 6% in glycogen. During the first gonotrophic cycle, about 35% of the labeled lipid stores were metabolized, with equal amounts being oxidized and transferred to the eggs. None of the other maternal stores labeled by fatty acid were metabolized during the first gonotrophic cycle. These results show that carbohydrate reserves are a critical source of energy during the first gonotrophic cycle, while lipid reserves are used equally for energy production and provisioning the eggs.     

Gluttony and sex in female ixodid ticks: how do they compare to other blood-sucking arthropods?

The central issue dealt with here is the role of copulation in the control of feeding behaviour in ticks and some haematophagous insects. Female ticks of the family Ixodidae normally engorge to approximately 100 x their unfed body weight, and then drop from the host, produce and lay eggs, and die. Virgins, on the other hand, normally do not exceed 5-40% (depending on species) of the normal engorged body weight. But instead of detaching voluntarily at that point most virgins remain fixed to the host for extended periods, waiting for males to find them so they can complete engorgement. Virgin haematophagous insects, and virgin ticks of the family Argasidae display little, if any, reduction in blood meal size compared to mated females, at least not during the first ovarian cycle. During subsequent ovarian cycles, meal size in some virgin insects may be somewhat reduced depending on how many eggs are retained in the reproductive tract, but the reduction is not nearly to the same extent as that observed for virgin ixodid females. The stimulatory effect of copulation on engorgement in the latter is caused by a pair of proteins (voraxin alpha and beta) produced in the testis and transferred to the female with the spermatophore. Here, I propose why it might be adaptive for an ixodid female to remain small until mated. The hypothesis is suggested from the facts that ixodid ticks remain attached to the host for days (rather than minutes), and that virgin ticks, above a certain critical weight, lose all opportunity for producing viable offspring should they be groomed off the host prematurely, or should the host die while ticks are still attached.     

Predatory mites avoid ovipositing near counterattacking prey

Attacking prey is not without risk; predators may endure counterattackby the prey. Here, we study the oviposition behaviour of a predatory mite(Iphiseius degenerans) in relation to its prey, thewesternflower thrips (Frankliniella occidentalis). This thrips iscapable of killing the eggs of the predator. Thrips and predatory mites - apartfrom feeding on each other - can also feed and reproduce on a diet of pollen.Because thrips may aggregate at pollen patches, such patches may be risky foroviposition by the predatory mites. We found that, in absence of thrips,predatory mites lay their eggs close to pollen, but further away when thripsarepresent. Predatory mite eggs near pollen were killed more frequently by thripsthan when they were deposited further away. The oviposition behaviour of thepredatory mite was also studied in absence of thrips, but in presence of thealarm pheromone of thrips. This pheromone is normally secreted upon contactwithpredators or competitors. When applied close to the pollen, predatory mitesoviposited significantly further away from it. When the alarm pheromone wasapplied away from the food source, most eggs were found near the pollen. Theseresults indicate that female predatory mites show flexible ovipositionbehaviourin response to the presence of their counterattacking prey.     

The role of free amino acids in osmoregulation of cricket blood (Acheta domesticus)

The blood volume increased during normal feeding, and did not decrease during fasting at the end of the stadium. The unexpectedly high blood volume of starved crickets might be an adaptation to increase chances for moulting via stretch receptor stimulation.The amount of blood amino acids was not changed by feeding, but increased with fasting or starvation. Thus amino acid levels in the blood were not directly related to amino acid input from the gut.The blood protein concentration did not change during starvation, but the amino acid concentration was 33% higher in starved crickets that drank water as opposed to those given saline to drink. Thus amino acid levels in the blood were not related directly to blood protein concentration.The blood amino acid concentration was 19–22 mM/l in response to salt intake by feeding crickets or starved crickets drinking saline. The concentration was 32–38 mM/l when the crickets were fasting prior to and after ecdysis or when starved with water to drink during the time when they would normally be feeding. The increase of amino acids during fasting was due to a proportional increase in all amino acids augmented by a 3 × increase in tyrosine. The increase during salt depleting starvation was due to a doubling of the two predominant amino acids proline and glycine. Proline and glycine were not increased in starved crickets drinking saline, thus starvation was not the reason for the increase. This is the first instance where specific amino acids have been implicated in osmoregulation.     

Infection of the malaria mosquito Anopheles gambiae with the entomopathogenic fungus Metarhizium anisopliae reduces blood feeding and fecundity

The entomopathogenic fungus Metarhizium anisopliae is being considered as a biocontrol agent against adult African malaria vectors. In addition to causing significant mortality, this pathogen is known to cause reductions in feeding and fecundity in a range of insects. In the present study we investigated whether infection with M. anisopliae affected blood feeding and fecundity of adult female malaria vectors Anopheles gambiae Giles sensu stricto. Mosquitoes were contaminated with either a low or a moderately high dose of oil-formulated conidia of M. anisopliae, and offered a single human blood meal 48, 72, or 96 h later to assess feeding propensity and individual blood meal size. In a second experiment, individual fungus-infected females were offered a blood meal every third day (to a total of 8 gonotrophic cycles), and allowed to oviposit after each cycle in order to quantify feeding propensity and fecundity. Infected females took smaller blood meals and displayed reduced feeding propensity. It was found that mosquitoes, inoculated with a moderately high dose of fungal conidia, exhibited reduced appetite related to increasing fungal growth. Of the fungus-infected females, the proportion of mosquitoes taking the second blood meal was reduced with 51%. This was further reduced to 35.3% by the 4th blood meal. During 8 feeding opportunities, the average number of blood meals taken by uninfected females was 4.39, against 3.40 (low dose), and 2.07 (high dose) blood meals for the fungus-infected females. Moreover, infected females produced fewer eggs per gonotrophic cycle and had a lower life-time fecundity. Epidemiological models show that both blood feeding and fecundity are among the most important factors affecting the likelihood of a mosquito transmitting malaria, which suggests that this fungus may have potential as biocontrol agent for vector-borne disease control.     

Lipophorin levels in the yellow fever mosquito,Aedes aegypti,and the effect of feeding

High density lipophorin (HDLp) is the major lipid transport vehicle in insect hemolymph. Using an indirect ELISA, levels of HDLp were measured in the yellow fever mosquito, Aedes aegypti. The level of lipophorin, when normalized to the total weight of the insect, was similar in the different developmental stages. Starvation (access to water only) of adult females did not affect the level of HDLp nor its density when compared to sugar-fed females. On the other hand, blood feeding (of normally sugar-fed females) resulted in a three-fold increase of the HDLp level at 40 h after feeding. This increase was accompanied by a slight but significant increase in the density of HDLp at 24 h after feeding. Ingestion of a lipid-free protein meal or a lipid-supplemented protein meal induced changes in HDLp level and density that were comparable to those induced by ingestion of a blood meal. Ingestion of a blood meal, following starvation (access to water only) from the moment of adult emergence, did not induce an increase in HDLp level. The results presented indicate that, in contrast to other insect species, A. aegypti responds to an increased need for lipid transport in the hemolymph by increasing the amount of HDLp. Arch. Insect Biochem. Physiol. 34:301–312, 1997. © 1997 Wiley-Liss, Inc.     

Modelling the effect of feeding-related mortality on the feeding strategy of tsetse (Diptera: Glossinidae)

Abstract. Free-living haematophagous insects risk death through host grooming responses or through increased susceptibility to predation whenever they take a bloodmeal. In this paper we investigate the effects of these risks on the feeding strategy of tsetse. A model is presented that allows for death of tsetse by starvation if they do not succeed in feeding within a fixed time (set at 6 days in the first instance) and for mortality specifically associated with feeding. In addition there is background mortality that applies to all flies at all times.
The model is used to compute the individual life-time fertility (number of female puparia per female) as a function of the probability of obtaining a meal (indicated by field data to be very high, usually > 0.85 per day) and the day on which flies start to search for a meal. We suggest that the feeding strategy that would be selected for is that which allows the maximum reproductive output. The model shows that this strategy involves making no attempts to feed for 3–4 days after the previous meal and then attempting to feed with the greatest possible probability until a meal is obtained. The predicted feeding interval, obtained independently of any trapping data, agrees closely with all previous estimates from field studies using a variety of methods. Preliminary results from a laboratory experiment reveal an increased risk of predation of recently fed as compared with hungry tsetse. The lower the actual feeding mortality the more frequently will flies be able to feed should conditions so demand. It is adaptive, however, for tsetse to delay attempting to feed for as long as they can, which is made possible by the near certainty of locating and feeding on a host within 1 day, using their sophisticated sensory systems.