Long-term regulation of sucrose intake by the adult Mediterranean fruit fly, Ceratitis capitata (Wiedemann)

Male and female medflies, Ceratitis capitata, were maintained for up to 8 days after emergence on 2% (58 mM), 8% (234 mM), or 16% (468 mM) sucrose solutions. Sucrose intake was recorded daily and whole-body analyses of lipid and glycogen were made at regular intervals.Greater volumes of fluid were imbibed by all flies on more dilute solutions. However, the amount of sucrose taken up over time was greatest for males on the 16% diet. Female intake was equal on the 8 and 16% diets.Males lost body triacylglycerol at a rate proportional to the dilution of the diet. Females on 8 and 16% sucrose lost no triacylglycerol throughout the 8-day period. Males and females, starved for 2 days after emergence lost 80% of their triacylglycerol and did not resume triacylglycerol synthesis when offered a diet rich in sucrose.     

Expression and regulation of vitellogenin messenger RNA in the mosquito,Aedes aegypti

Vitellogenin (yolk protein) gene expression in the mosquito was investigated at the level of mRNA using a subcloned fragment (403-1c) of the vitellogenin DNA derived from an Aedes aegypti genomic library. Message appeared 1–3 hr after a blood meal, peaked at 36 hr and was rapidly degraded thereafter. Fluctuations in levels of 20-hydroxyecdysone after a blood meal coincided with accumulation of vitellogenin message. Blood-fed, decapitated females injected with 5 μg of 20-hydroxyecdysone accumulated up to 75% of the message found in blood-fed controls. Fat bodies from non-blood-fed females incubated with physiological levels of 20-hydroxyecdysone and the juvenile hormone analog methoprene contained twice as much vitellogenin message as those incubated with 20-hydroxyecdysone alone. Methoprene alone had no effect.     

Toward a description of the sialome of the adult female mosquito Aedes aegypti

To describe the set of mRNA and protein expressed in the salivary glands (sialome) of Aedes aegypti mosquitoes, we randomly sequenced a full-length cDNA library of this insect and performed Edman degradation of PVDF-transferred protein bands from salivary homogenates. We found 238 cDNA clusters which contained those coding for 10 of the 11 proteins found by aminoterminal degradation. All six previously described salivary proteins were found in this library. Full-length sequences of 32 novel cDNA sequences are reported, one of which is the product of a transposable element. Among the 31 novel protein sequences are 4 additional members of the D7 protein family; 4 novel members of the antigen 5 family (a protein family not reported in Aedes); a novel serpin; a novel member of the 30-kDa allergen of Ae. Aegypti; a secreted calreticulin; 2 proteins similar to mammalian angiopoietins; adenosine deaminase; purine hydrolase; lysozyme; a C-type lectin; 3 serine proteases, including one with high similarity to Bombyx prophenoloxidase activating enzyme; 2 proteins related to invertebrate immunity; and several sequences that have no significant matches to known proteins. The possible role of these proteins in blood and sugar feeding by the mosquito is discussed.     

Nectar feeding by the early-spring mosquito Aedes provocans

Abstract. Nectar feeding by males and females of the mosquito Aedes provocans was studied at a site near Belleville, Ontario, Canada. Canada plum, Prunus nigra , and especially pin cherry, P. pensylvanica , bloomed contemporaneously with the emergence of Ae. provocans and were important nectar sources for adult mosquitoes during their first week of life. Blossoms of P. pensylvanica shielded for 24 h from foragers produced an average of 0.14 mg of sugar (≅ 2.3 J). This nectar was avidly sought by both sexes of Ae.provocans; > 97% of the blossoms were visited by mosquitoes in the first few days of blooming. Young adult mosquitoes were found on blossoms at all hours of the day and night; feeding on P. nigra was strongly eocrepuscular, whereas on P.pensylvanica feeding was much less strongly periodic. Adults foraged for nectar in an energy-conserving, pedestrian strategy, devoting 56% (females) and 68% (males) of their time on blossoms to nectar feeding during foraging bouts that lasted a median of 5.3min. Both sexes sought nectar soon after emergence - males before they had completed hypopygial rotation or swarmed, and females before mating or host seeking. Female Ae.provocans sought nectar in all stages of oogenesis, but primarily at the initiation of a gonotrophic cycle. Energy stores in the crop averaged 18 J per female, with a distribution that depended on gonotrophic age and parity.     

Internal regulation of rate of digestion of blood meals in the mosquito, Aedes aegypti.

Rate of digestion of blood meals proceeded more rapidly in females of Aedes aegypti that were inseminated or injected with matrone (extract of male accessory glands) than in virgin females. Digestion rate was determined by interfacial precipitin tests and immunodiffusion tests for undigested blood-meal proteins. Application of a cervical ligature within 1 hr of blood feeding retarded the digestion rate. Ligated females that received brain transplants from blood-fed donors digested their blood meals at a rate similar to that of unligated controls. When ligatures were not applied until 12 hr after feeding no delay in digestion was observed.     

Role of the fat body in the regulation of host-seeking behaviour in the mosquito,Aedes aegypti

Following a blood meal that initiates oöcyte development, the host-seeking behaviour of Aedes aegypti mosquitoes is inhibited by a haemolymph-borne factor that is released in response to a humoral signal from a vitellogenic ovary. This inhibition is accompanied by a decrease in the sensitivity of the peripheral lactic acid receptors. Implantation of corpora allata, medial neurosecretory cells, or terminal abdominal ganglia from blood-fed donors could not induce the inhibition in sugar-fed recipients. However, fat body transplanted from blood-fed into sugar-fed females suppressed host-seeking behaviour as well as the sensitivity of lactic acid receptors, suggesting that the source of the behavioural inhibitor is the fat body. Resting-stage ovaries from other mosquito species inhibited host-seeking after the A. aegypti host was fed on blood only if the fat body was activated by the donor ovary.     

Distention-mediated egg maturation in the mosquito,Aedes aegypti

Abdominal distention accelerates the release of a factor from the head of blood-fed Aedes aegypti mosquitoes. The critical period during which the head is required for oögenesis following blood ingestion is approx 6 h with a 5 μl meal, but small blood meals of 1 μl require the head to be present for significantly longer. Increasing the abdominal distention by supplementing the 1 μl meal with saline results in a critical period similar to that with 5 μl of blood. The information from the distended abdomen appears to travel via the ventral nerve cord. Transection of the ventral nerve cord prevents oögenesis from occurring after small blood meals, but not with larger blood volumes. Topical application of 100 pg of juvenile hormone III can substitute for the distention message.     

Osmoregulation in larvae of the salt-marsh mosquito, Aedes taeniorhynchus

Larvae of Aedes taeniorhynchus (Wiedemann) were reared in media with salinities from that of distilled water up to and including 300% of that of sea water to investigate certain aspects of their potential physiological range. Regulation of hemolymph osmotic pressure and chloride ions was also studied.Larvae showed normative growth rate in all concentrations from distilled water to 150% sea water (SW), but in salinities between 150% to 300% growth was retarded. Hemolymph osmotic pressure and hemolymph chloride were both hyper-and hypoosmotically regulated. Anal papillae size was inversely related with increased concentration of the sea water medium, e.g., from 443×142 in distilled water to 116×62 in 100% SW. The average hemolymph osmotic pressure was higher in fed larvae than in starved larvae. Hemolymph osmotic pressure increased for 7 hr before equilibrating with the medium when larvae reared to the 4th instar in 10% SW were transferred to 100% SW, whereas larvae reared in 100% SW and transferred to 10% SW showed a decrease in hemolymph osmotic pressure before equilibrating. Regulation of hemolymph chloride was found to be a function of the anal papillae, as chloride levels dropped significantly in larvae with chemically cauterized anal papillae when they were maintained in lower concentrations. It is suggested that the limitations of A. taeniorhynchus larvae primarily to salt-marshes are not due to an inability to survive and grow successfully in fresh water, but due to other ecological interactions.

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Zusammenfassung Larven von Aedes taeniorhynchus wurden in Medien mit einer Salinität von a.ddest. bis zu der von 300% Meerwasser (MW) gehalten, um die folgenden Aspekte einer möglichen physiologischen Wirkung zu untersuchen: a) Überleben und Wachstum der Larven, b) Osmotischer Druck der Hämolymphe (HL), sowie Grösse der Analpapillen in Abhängigkeit vom Zuchtmedium, c) Wirkung von Fütterung und Hungern auf den osmotischen Druck der HL, d) Wirkung der Übertragung von niedrigerer zu höherer Salinität und umgekehrt auf den osmotischen Druck und die Analpapillengrösse und e) Regulation des Chloridions in der HL.Osmotischer Druck der HL wurde bestimmt mit Hilfe des Mikrocryoskops, die Chloridio-nenkonzentration der HL durch Ultramikro-Volhard-Titration.Die Larven zeigten normales Wachstum und normale Überlebensrate bei allen Konzentrationen von a. dest. bis 150% MW, zwischen 150% und 300% MW war das Wachstum verzögert. Osmotischer Druck der HL und Chlorid der HL waren hyperosmotisch reguliert bis 10% MW und hypotonisch zwischen 25% und 300% MW. Die Grösse der Analpapillen nahm mit zunehmender Konzentration des MW-Mediums ab, z. B. von 443×142 m in a. dest. auf 116×62 m in 100% MW. Der durchschnittliche osmotische Druck der HL war bei gefütterten Larven höher als bei gehungerten. Wenn Larven, die bis zum 4. Stadium in 10% MW gehalten wurden, in 100% MW übertragen wurden, stieg der osmotische Druck der HL weit über die für 100% MW festgestellte Gleichgewichtslage hinaus an und näherte sich dieser (also durch Abnahme) erst nach 7 Stunden; bei Übertragung von 100% auf 10% MW erfolgte entsprechend zunächst eine übernormale Abnahme des osmotischen Drucks der HL vor Erreichen des Gleichgewichts. Die Regulation der HL-Chloride erwies sich als eine Funktion der Analpapillen, da der Chloridspiegel signifikant abfiel bei Larven, die mit chemisch kautorisierten Analpapillen in niedrigen Konzentrationen gehalten wurden. Es ist anzunehmen, dass die Beschränkung des Vorkommens von Aedes taeniorhynchus-Larven auf Aussenmarschen nicht verursacht wird durch die Unfähigkeit, in Süsswasser zu überleben und sich zu entwickeln, sondern durch andere ökologische Einflüsse.

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Contribution no. 315, Florida State Division of Health, Florida Medical Entomology Laboratory, P.O. Box 520, Vero Beach, Florida 32960.     

The salivary purine nucleosidase of the mosquito,Aedes aegypti

A cDNA clone originating from adult female Aedes aegypti mosquitoes was found with substantial similarity to nucleosidases of the EC 3.2.2.1 enzyme class. Although this type of enzyme is unusual in animals, abundant enzyme activity was found in salivary homogenates of this mosquito, but not in salivary homogenates of the mosquitoes Anopheles gambiae and Culex quinquefasciatus, or the sand fly Lutzomyia longipalpis. Aedes salivary homogenate hydrolyses inosine and guanosine to hypoxanthine and xanthine plus the ribose moiety, but does not hydrolyse the pyrimidines uridine and cytidine, thus characterizing the presence of a purine nucleosidase activity. The enzyme is present in oil-induced saliva, indicating that it is secreted. Male Ae. aegypti salivary gland homogenates (SGH) have very low purine nucleosidase activity, suggesting that the enzyme plays a role in mosquito blood feeding. A novel isocratic HPLC method to separate nucleosides and their bases is described.     

Activation in vitro of vitellogenin uptake by the oocytes of the mosquito, Aedes aegypti

A blood meal stimulates the ovaries of the mosquito, Aedes aegypti , to internalize vitellogenin from the haemolymph for storage as yolk. The nature of the stimulation is biphasic, with a pulsed factor from the head released within the first 5 min of feeding stimulating the first phase of uptake, and the continuous release of another head factor initiating the second phase. Using an in vitro assay of [³⁵S]-vitellogenin uptake by ovaries, we sought to determine the intracellular signalling pathways utilized by the ovaries during stimulation of both phases of uptake. The adenylate cyclase activators forskolin and ethanol produced a statistically significant increase in rate of uptake in previtellogenic ovaries to levels characteristic of first-phase activated ovaries. Forskolin also induced the second phase of uptake in ovaries that had been arrested in the first phase of uptake by decapitation of the mosquito within 5 min of ingesting blood. Initiation and up-regulation of the rate of vitellogenin uptake does not depend on new protein synthesis.     

FLP-mediated recombination in the vector mosquito, Aedes aegypti.

The activity of a yeast recombinase, FLP, on specific target DNA sequences, FRT, has been demonstrated in embryos of the vector mosquito, Aedes aegypti. In a series of experiments, plasmids containing the FLP recombinase under control of a heterologous heat-shock gene promoter were co-injected with target plasmids containing FRT sites into preblastoderm stage mosquito embryos. FLP-mediated recombination was detected between (i) tandem repeats of FRT sites leading to the excision of specific DNA sequences and (ii) FRT sites located on separate plasmids resulting in the formation of heterodimeric or higher order multimeric plasmids. In addition to FRT sites originally isolated from the yeast 2 microns plasmid, a number of synthetic FRT sites were also used. The synthetic sites were fully functional as target sites for recombination and gave results similar to those derived from the yeast 2 microns plasmid. This successful demonstration of yeast FLP recombinase activity in the mosquito embryo suggests a possible future application of this system in establishing transformed lines of mosquitoes for use in vector control strategies and basic studies.