Sugar Meal Research Articles

Species identification of plant tissues from the gut of An. sergentii by DNA analysis

There are three commonly used assays to identify plant material in insect guts: the cold anthrone test for fructose, the cellulose staining test for visualizing plant tissue and gas chromatography for seeking unique sugar content profiles. Though sugar and cellulose tests can distinguish between the general sources of sugar meal (nectar versus tissue), they cannot identify the species of plant sources. Even gas chromatography profiles can be problematic; there are reported instances of intra-specific variation as well as inter-specific and intergeneric variation that can mar results. Here, we explore the potential for DNA analysis to help resolve this issue. First, Anopheles sergentii were exposed to branches of two species of highly attractive flowering bushes in the laboratory and the great majority (∼90–98%) were positive for sugar from nectar while very few were positive for cellulose (∼0.5–8%) and DNA (6–19%). Moreover, laboratory An. sergentii showed opposing preferences, tending to obtain sugar from nectar of one plant ( Tamarix nilotica) but to feed more on tissue from the other ( Ochradenus baccatus). An. sergentii are exposed to a wide variety of plants in their natural desert habitats and in the absence of flowers in the dry season, they resort to feeding specifically on tissues of a few plants. According to DNA analysis the favorite plants were Suaeda asphaltica, Malva nicaeensis and Conyza dioscoridis, which are succulents that account for less than 1% of vegetation in the area.

Effects of Melezitose and Stachyose on Adult Longevity and Virus Persistence inCulicoides sonorensis(Diptera: Ceratopogonidae)

A wide variety of blood-feeding Diptera feed on extrafloral sugar sources such as homopteran honeydew. The significance of these sugar sources to insect survival and disease transmission is poorly known. Culicoides sonorensis, a biting midge, can survive on plant sugars but might also feed on homopteran honeydew. The survival of C. sonorensis fed 10% melezitose or 10% stachyose was compared to that of C. sonorensis fed 10% sucrose. A significantly greater number of C. sonorensis survived longer when fed melezitose than when fed either stachyose or sucrose. The effect of sugar meals on vector competence was measured using bluetongue and epizootic hemorrhagic disease viruses. There was no significant difference in viral persistence. However, if C. sonorensis feeds on homopteran honeydew in the wild, a larger number of midges could survive to transmit orbiviruses.

Honeydew and nectar sugars differentially affect flight performance in female black flies

In the wild, black flies (Diptera: Simuliidae) feed on both floral nectar and homopteran honeydew. We tested the hypothesis that sugar meal composition affects flight performance using tethered black flies that had been flown to exhaustion. Individuals were then fed distilled water (dH2O), 10% (m/v) artificial nectar (AN), or 20% (m/v) artificial honeydew (AH) sugars, and the distance, speed, and duration of the resultant flights were recorded using a computerized flight mill. Following a dH2O meal, flies were unable to regain flight. There were three different artificial honeydew diets (AHlow, AHmed, AHhigh), each differing in the proportion and composition of simple sugars (glucose, fructose, sucrose) and complex sugars (melezitose, stachyose). Flies flew the farthest and longest on AH meals that contained higher levels of complex sugars (AHmed, AHhigh) compared with flies fed on AN (simple sugars only) or AHlow. On this flight mill, the mean flight speed was 10.6 ± 3.5 m/min and was not affected by the type of sugar meal ingested.

Asaia, a versatile acetic acid bacterial symbiont, capable of cross‐colonizing insects of phylogenetically distant genera and orders

Bacterial symbionts of insects have been proposed for blocking transmission of vector-borne pathogens. However, in many vector models the ecology of symbionts and their capability of cross-colonizing different hosts, an important feature in the symbiotic control approach, is poorly known. Here we show that the acetic acid bacterium Asaia, previously found in the malaria mosquito vector Anopheles stephensi, is also present in, and capable of cross-colonizing other sugar-feeding insects of phylogenetically distant genera and orders. PCR, real-time PCR and in situ hybridization experiments showed Asaia in the body of the mosquito Aedes aegypti and the leafhopper Scaphoideus titanus, vectors of human viruses and a grapevine phytoplasma respectively. Cross-colonization patterns of the body of Ae. aegypti, An. stephensi and S. titanus have been documented with Asaia strains isolated from An. stephensi or Ae. aegypti, and labelled with plasmid- or chromosome-encoded fluorescent proteins (Gfp and DsRed respectively). Fluorescence and confocal microscopy showed that Asaia, administered with the sugar meal, efficiently colonized guts, male and female reproductive systems and the salivary glands. The ability in cross-colonizing insects of phylogenetically distant orders indicated that Asaia adopts body invasion mechanisms independent from host-specific biological characteristics. This versatility is an important property for the development of symbiont-based control of different vector-borne diseases.

GLUT2 mutations, translocation, and receptor function in diet sugar managing

Cloned 20 years ago, GLUT2 is a facilitative glucose transporter in the liver, pancreas, intestine, kidney, and brain. It ensures large bidirectional fluxes of glucose in and out the cell due to its low affinity and high capacity. It also transports other dietary sugars, such as fructose and galactose, within the range of physiological concentrations. Sugars and hormones regulate its gene expression. The contribution of GLUT2 to human metabolic diseases previously appeared modest. However, in the past decade, three major features of the GLUT2 protein have been revealed. First, GLUT2 mutations cause the severe but rare Fanconi-Bickel syndrome, mainly characterized by glycogenosis. Recently, a GLUT2 polymorphism has been associated with preferences for sugary food. Second, the GLUT2 location at the cell surface is regulated; this governs cellular activities dependent on glucose in the intestine and possibly those in the liver and pancreas. For instance, GLUT2 translocation from an intracellular pool to the apical membrane after a sugar meal transiently increases sugar uptake by enterocytes (reviewed in 32). Third, GLUT2 functions as a membrane receptor of sugar. Independently of glucose metabolism, GLUT2 detects the presence of extracellular sugar and transduces a signal to modulate cell functions, including beta-cell insulin secretion, renal reabsorption, and intestinal absorption according to the sugar environment. These recent developments are examined here in heath and metabolic disease, highlighting various unanswered questions.

The salivary gland transcriptome of the neotropical malaria vector Anopheles darlingi reveals accelerated evolution of genes relevant to hematophagy

BackgroundMosquito saliva, consisting of a mixture of dozens of proteins affecting vertebrate hemostasis and having sugar digestive and antimicrobial properties, helps both blood and sugar meal feeding. Culicine and anopheline mosquitoes diverged ~150 MYA, and within the anophelines, the New World species diverged from those of the Old World ~95 MYA. While the sialotranscriptome (from the Greek sialo, saliva) of several species of the Cellia subgenus of Anopheles has been described thoroughly, no detailed analysis of any New World anopheline has been done to date. Here we present and analyze data from a comprehensive salivary gland (SG) transcriptome of the neotropical malaria vector Anopheles darlingi (subgenus Nyssorhynchus).ResultsA total of 2,371 clones randomly selected from an adult female An. darlingi SG cDNA library were sequenced and used to assemble a database that yielded 966 clusters of related sequences, 739 of which were singletons. Primer extension experiments were performed in selected clones to further extend sequence coverage, allowing for the identification of 183 protein sequences, 114 of which code for putative secreted proteins.ConclusionComparative analysis of sialotranscriptomes of An. darlingi and An. gambiae reveals significant divergence of salivary proteins. On average, salivary proteins are only 53% identical, while housekeeping proteins are 86% identical between the two species. Furthermore, An. darlingi proteins were found that match culicine but not anopheline proteins, indicating loss or rapid evolution of these proteins in the old world Cellia subgenus. On the other hand, several well represented salivary protein families in old world anophelines are not expressed in An. darlingi.

Methylparaben in Anopheles gambiae s.l. sugar meals increases longevity and malaria oocyst abundance but is not a preferred diet

The antimicrobial and antifungal chemical methylparaben (methyl-4-hydroxybenzoate) was added to the adult sucrose diet of Anopheles gambiae and Anopheles arabiensis, and its effect on longevity was determined. In all cases, significant increases in longevity were observed when 0.2% (w/v) methylparaben was added to meals that were refreshed weekly. When fresh sugar diet was refreshed daily, no increase in longevity was observed due to methylparaben suggesting that the effect of methylparaben is to preserve the quality of the sugar diet. No longevity effect of providing pure water in addition to sugar- or methylparaben-supplemented meals was observed. Feeding preference tests were performed to determine whether meals containing methylparaben were preferred, and whether, when given no choice but the less-preferred diet, mosquitoes would consume less sugar. Using the stable carbon isotope 13C in paired tests, we show that the sugar diet containing methylparaben was clearly avoided by A. gambiae but not A. arabiensis. Little effect of methylparaben on the total amount of sugar consumed was observed when mosquitoes were given no diet choice. Methylparaben effects on Plasmodium cynomolgi B oocyst formation and encapsulation were observed in a normal A. gambiae stock and one which encapsulates at a high frequency. Nearly two-fold increases in the number of both normal and encapsulated oocysts were observed as a result of methylparaben in the diet. Because of its longevity effects, we have implemented methylparaben use for all mosquitoes in our holdings and recommend it as a routine sugar meal supplement.

High nocturnal CO2 emanation guides the sand fly Phlebotomus papatasi to sugar‐rich plants

Abstract In arid areas, Phlebotomus papatasi obtains essential carbohydrates by feeding on green tissues of plants. There is a great variation in sugar content, metabolic rates and dark respiration between conspecific plants, and also between branches and leaves of the same plant. It is hypothesized that high nocturnal release of CO2 may guide the sand flies to rich sugar sources. Comparisons of Ph. papatasi feeding on branches of different plant species demonstrate a significant positive correlation between the level of sugar in leaves, the rate of sand fly feeding on them and the size of ingested sugar meals. Mean nocturnal CO2 emanation of low‐sugar Ricinus communis branches is 26.5 ppm and that of sugar rich branches is 86.3 ppm above room level. Low‐sugar Capparis spinosa branches release 45.0 ppm and the emission from sugar rich branches is 76.0 ppm above room level. Branches with similar emissions, placed behind net partitions, are used in no‐choice orientation experiments: R. communis branches releasing high CO2 levels are approached by 53.7% of the flies compared with 3.0% of flies that orientate to low CO2 branches. Capparis spinosa with high CO2 emission are approached by 9.8% of the flies compared with 0.95% of flies that orientate to low CO2 branches.

Identification, expression and characterisation of a major salivary allergen (Cul s 1) of the biting midge Culicoides sonorensis relevant for summer eczema in horses

Salivary proteins of Culicoides biting midges are thought to play a key role in summer eczema (SE), a seasonal recurrent allergic dermatitis in horses. The present study describes the identification, expression and clinical relevance of a candidate allergen of the North American midge Culicoides sonorensis. Immunoblot analysis of midge saliva revealed a 66kDa protein (Cul s 1) that was bound by IgE from several SE-affected (SE+) horses. Further characterisation by fragmentation, mass spectrometry and bioinformatics identified Cul s 1 as maltase, an enzyme involved in sugar meal digestion. A cDNA encoding Cul s 1 was isolated and expressed as a polyhistidine-tagged fusion protein in a baculovirus/insect cell expression system. The clinical relevance of the affinity-purified recombinant Cul s 1 (rCul s 1) was investigated by immunoblotting, histamine release testing (HRT) and intradermal testing (IDT) in eight SE+ and eight control horses. Seven SE+ horses had rCul s 1-specific IgE, whereas only one control animal had IgE directed against this allergen. Furthermore, the HRT showed rCul s 1 induced basophil degranulation in samples from seven of eight SE+ horses but in none of the control animals. rCul s 1 also induced immediate (7/8), late-phase (8/8) and delayed (1/8) skin reactivity in IDT on all SE+ horses that had a positive test with the whole body extract (WBE) of C. sonorensis. None of the control horses showed immediate or delayed skin reactivity with rCul s 1, and only one control horse had a positive late-phase response, while several non-specific late-phase reactions were observed with the insect WBE. Thus, we believe rCul s 1 is the first specific salivary allergen of C. sonorensis to be described that promises to advance both in vitro and in vivo diagnosis and may contribute to the development of immunotherapy for SE in horses.

An insight into the sialotranscriptome of the non-blood feeding Toxorhynchites amboinensis mosquito

All adult mosquitoes take sugar meals, and most adult females also take blood meals to develop eggs. Salivary glands (SG) of males are thus much smaller and do not contain many of the antihemostatic and antiinflammatory compounds found in females. In the past 5 years, transcriptome analyses have identified nearly 70 different genes expressed in adult female SG. For most of these, no function can be assigned in either blood or sugar feeding. Exceptionally, Toxorhynchites mosquitoes are unusual in that they never feed on blood, and the SG of adults are identical in both sexes. Transcriptome analysis of the adult SG of this mosquito was performed to increase knowledge of the evolution of blood feeding—and to identify polypeptide families associated with sugar feeding—in mosquitoes.

Insulin Internalizes GLUT2 in the Enterocytes of Healthy but Not Insulin-Resistant Mice

A physiological adaptation to a sugar-rich meal is achieved by increased sugar uptake to match dietary load, resulting from a rapid transient translocation of the fructose/glucose GLUT2 transporter to the brush border membrane (BBM) of enterocytes. The aim of this study was to define the contributors and physiological mechanisms controlling intestinal sugar absorption, focusing on the action of insulin and the contribution of GLUT2-mediated transport. The studies were performed in the human enterocytic colon carcinoma TC7 subclone (Caco-2/TC7) cells and in vivo during hyperinsulinemic-euglycemic clamp experiments in conscious mice. Chronic high-fructose or high-fat diets were used to induce glucose intolerance and insulin resistance in mice. In Caco-2/TC7 cells, insulin action diminished the transepithelial transfer of sugar and reduced BBM and basolateral membrane (BLM) GLUT2 levels, demonstrating that insulin can target sugar absorption by controlling the membrane localization of GLUT2 in enterocytes. Similarly, in hyperinsulinemic-euglycemic clamp experiments in sensitive mice, insulin abolished GLUT2 (i.e., the cytochalasin B-sensitive component of fructose absorption), decreased BBM GLUT2, and concomitantly increased intracellular GLUT2. Acute insulin treatment before sugar intake prevented the insertion of GLUT2 into the BBM. Insulin resistance in mice provoked a loss of GLUT2 trafficking, and GLUT2 levels remained permanently high in the BBM and low in the BLM. We propose that, in addition to its peripheral effects, insulin inhibits intestinal sugar absorption to prevent excessive blood glucose excursion after a sugar meal. This protective mechanism is lost in the insulin-resistant state induced by high-fat or high-fructose feeding.

Glycolytic and chitinolytic activities of Phlebotomus papatasi (Diptera: Psychodidae) from diverse ecological habitats

The sand fly Phlebotomus papatasi Scopoli, 1786, the vector of Leishmania major Yakimoff et Schokhor, 1914, is found in desert areas where sugars are scarce but also in habitats that abound in sugar sources. The sand flies require sugar meals from plant sources for their energy requirements and to hydrolyze these complex sugars, they need a repertoire of glycosidases. We presumed that there are differences in the levels of glycosidase activities in flies from such habitats and also assumed that they may be instrumental in modulating the flies' susceptibility to L. major infections. Phlebotomus papatasi originating from diverse ecological habitats ranging from an oasis to desert sites were colonized. They were analyzed for weight changes and glycosidase activities before and after feeding on 1M sucrose solution. Oasis flies were smaller than desert flies but took larger sugar meals. Homogenates of these flies hydrolyzed 16 synthetic and 2 natural glycoside substrates to varying degrees. The arid-region flies tended to produce more glycosidase activity than those originating in sugar-rich environments, especially sucrase, alpha- and beta-glucosidase, aalpha-fucosidase, alpha-mannosidase, and alpha- and beta-N-acetylgalactosaminidase. However, chitinolytic enzyme activities and particularly the beta-N-acetylhexosaminidase activity of oasis flies were higher than other flies tested. In comparing the desert flies, there were also significant differences in glycolytic enzyme activities between the spring-line (flowering season) of flies and the autumn-line (end of dry season) flies. A range of saccharide inhibitors was tested to demonstrate the specificity of the enzymes.

Frequent Occurrence of Rapid as Well as Delayed Gastric Emptying of a Corn Flakes and Milk Meal in Clinical Patients With Gastrointestinal Symptoms

Early satiety, nausea and vomiting have traditionally been associated with delayed gastric emptying (GE). A study was performed to determine the frequency of rapid/delayed GE in 100 patients sequentially referred for scintigraphic GE using a corn flakes and milk meal. A retrospective review of 100 consecutive GE studies at the University Hospital, San Antonio, Texas, was performed. Each patient received a semisolid meal containing corn flakes, milk, and sugar (200 kcal, 6 g fat, 7 g protein, and 30 g carbohydrates) and 37.0 MBq (1 mCi) of Tc-99m sulfur colloid according to a standard clinical protocol followed by dynamic 1-minute planar acquisitions for 60 minutes. Gastric emptying times were classified based on the 50% emptying time as follows: 30 to 60 minutes for normal, abnormally delayed as >60 minutes, and abnormally rapid as <30 minutes. Twenty-eight patients demonstrated rapid GE, 25 delayed GE, and 45 normal GE. Fifteen (54%) patients with rapid GE were diabetic, 4 (14%) had impaired fasting glucose values, and 9 (32%) were normoglycemic. Fourteen (56%) patients with delayed GE were diabetic, one (4%) had impaired fasting glucose, and 10 (40%) were normoglycemic. Both patients with delayed and those with rapid GE had nausea as the most common symptom followed by early satiety (rapid GE) and vomiting (delayed GE). Of 28 patients with rapid GE, 26 were on promotility agents. The number of patients with rapid GE of the corn flakes, milk, and sugar meal is appreciably greater (28%) than previously reported with other meals. This relative large number is likely related to the meal composition and the homogeneous dispersal of the label within the meal.

An annotated catalogue of salivary gland transcripts in the adult female mosquito, Ædes ægypti*

BackgroundSaliva of blood-sucking arthropods contains a cocktail of antihemostatic agents and immunomodulators that help blood feeding. Mosquitoes additionally feed on sugar meals and have specialized regions of their glands containing glycosidases and antimicrobials that might help control bacterial growth in the ingested meals. To expand our knowledge on the salivary cocktail of Ædes ægypti, a vector of dengue and yellow fevers, we analyzed a set of 4,232 expressed sequence tags from cDNA libraries of adult female mosquitoes.ResultsA nonredundant catalogue of 614 transcripts (573 of which are novel) is described, including 136 coding for proteins of a putative secretory nature. Additionally, a two-dimensional gel electrophoresis of salivary gland (SG) homogenates followed by tryptic digestion of selected protein bands and MS/MS analysis revealed the expression of 24 proteins. Analysis of tissue-specific transcription of a subset of these genes revealed at least 31 genes whose expression is specific or enriched in female SG, whereas 24 additional genes were expressed in female SG and in males but not in other female tissues. Most of the 55 proteins coded by these SG transcripts have no known function and represent high-priority candidates for expression and functional analysis as antihemostatic or antimicrobial agents. An unexpected finding is the occurrence of four protein families specific to SG that were probably a product of horizontal transfer from prokaryotic organisms to mosquitoes.ConclusionOverall, this paper contributes to the novel identification of 573 new transcripts, or near 3% of the Æ. ægypti proteome assuming a 20,000-protein set, and to the best-described sialome of any blood-feeding insect.

Associative learning of odor with food- or blood-meal by Culex quinquefasciatus Say (Diptera: Culicidae)

The ability of many insects to learn has been documented. However, a limited number of studies examining associative learning in medically important arthropods has been published. Investigations into the associative learning capabilities of Culex quinquefasciatus Say were conducted by adapting methods commonly used in experiments involving Hymenoptera. Male and female mosquitoes were able to learn a conditioned stimulus that consisted of an odor not normally encountered in nature (synthetic strawberry or vanilla extracts) in association with an unconditioned stimulus consisting of either a sugar (males and females) or blood (females) meal. Such information could lead to a better understanding of the ability of mosquitoes to locate and select host and food resources in nature.

The sialotranscriptome of adult male Anopheles gambiae mosquitoes

Adult mosquitoes feed on sugary meals to obtain energy for flight and other activities, while anautogenous females take a blood meal to develop eggs. Accordingly, female but not male salivary glands possess several antihemostatic components to facilitate acquisition of blood, while both sexes have activities related to digestion of the sugar meal as well as antimicrobials to maintain meal integrity. Studies on adult female sialotranscriptomes indicated a set of ∼70 proteins and peptides possibly secreted in saliva that presumably facilitate sugar and blood meals. Most of these proteins have no known function, so no assignment to blood or sugar feeding is possible. Microarray and RT-PCR studies attempted to identify sex specificity of these transcripts. Our present study complements the previous data set, comparing ∼1000 randomly sequenced clones of a male adult salivary gland cDNA library with the female set. Statistically significant differences were found in 16 transcripts found exclusively in the female library, 4 transcripts significantly female enriched but also found in male glands, and 6 transcripts enriched in male glands. We additionally found a transcript in male salivary glands with a trypsin inhibitor-like (TIL) domain that we presume codes for an antimicrobial peptide; a novel defensin transcript was also found in the male sialotranscriptome. Supplemental tables can be found at http://www.ncbi.nlm.nih.gov/projects/omes/

The effects of blood feeding and exogenous supply of tryptophan on the quantities of xanthurenic acid in the salivary glands of Anopheles stephensi (Diptera: Culicidae)

Xanthurenic acid (XA), produced as a byproduct during the biosynthesis of insect eye pigment (ommochromes), is a strong inducer of Plasmodium gametogenesis at very low concentrations. In previous studies, it was shown that XA is present in Anopheles stephensi (Diptera: Culicidae) mosquito salivary glands and that during blood feeding the mosquitoes ingested their own saliva into the midgut. Considering these two facts together, it is therefore likely that XA is discharged with saliva during blood feeding and is swallowed into the midgut where it exerts its effect on Plasmodium gametocytes. However, the quantities of XA in the salivary glands and midgut are unknown. In this study, we used high performance liquid chromatography with electrochemical detection to detect and quantify XA in the salivary glands and midgut. Based on the results of this study, we found 0.28 ± 0.05 ng of XA in the salivary glands of the mosquitoes, accounting for 10% of the total XA content in the mosquito whole body. The amounts of XA in the salivary glands reduced to 0.13 ± 0.06 ng after mosquitoes ingested a blood meal. Approximately 0.05 ± 0.01 ng of XA was detected in the midgut of nonblood fed An. stephensi mosquitoes. By adding synthetic tryptophan as a source of XA into larval rearing water (2 mM) or in sugar meals (10 mM), we evaluated whether XA levels in the mosquito (salivary glands, midgut, and whole body) were boosted and the subsequent effect on infectivity of Plasmodium berghei in the treated mosquito groups. A female specific increase in XA content was observed in the whole body and in the midgut of mosquito groups where tryptophan was added either in the larval water or sugar meals. However, XA in the salivary glands was not affected by tryptophan addition to larval water, and surprisingly it reduced when tryptophan was added to sugar meals. The P. berghei oocyst loads in the mosquito midguts were lower in mosquitoes fed tryptophan treated sugar meals than in mosquitoes reared on tryptophan treated larval water. Our results suggest that mosquito nutrition may have a significant impact on whole body and midgut XA levels in mosquitoes. We discuss the observed parasite infectivity results in relation to XA’s relationship with malaria parasite development in mosquitoes.

Sucrose sham feeding on a binge schedule releases accumbens dopamine repeatedly and eliminates the acetylcholine satiety response

Drinking a sugar solution on an intermittent schedule can promote sugar bingeing and cause signs of dependence while releasing dopamine repeatedly like a drug of abuse. It is hypothesized that sweet taste alone is sufficient for this effect in sucrose bingeing rats. On the theory that acetylcholine in the nucleus accumbens plays a role in satiety, it is further hypothesized that purging the stomach contents will delay acetylcholine release. Rats with gastric fistulas and nucleus accumbens guide shafts for microdialysis were fed 12 h each day. During the first hour, fistulas were open for the sham-feeding group and closed for the real-feeding group, and 10% sucrose was the only food source. For the remaining 11 h, liquid rodent diet was available as well as the 10% sucrose to provide a balanced diet. In microdialysis tests during the first sugar meal on days 1, 2 and 21, extracellular dopamine increased at least 30% each day in both groups. Acetylcholine also increased during the sugar meals for the real-feeding animals, but not during sham feeding. In conclusion, the taste of sugar can increase extracellular dopamine in the nucleus accumbens without fail in animals on a dietary regimen that causes bingeing and sugar dependency. During sham feeding, the acetylcholine satiation signal is eliminated, and the animals drink more. These findings support the hypothesis that dopamine is released repeatedly in response to taste when bingeing on sweet food, and the acetylcholine satiety effect is greatly reduced by purging; this may be relevant to bulimia nervosa in humans.

An updated catalogue of salivary gland transcripts in the adult female mosquito,Anopheles gambiae

Salivary glands of blood-sucking arthropods contain a variety of compounds that prevent platelet and clotting functions and modify inflammatory and immunological reactions in the vertebrate host. In mosquitoes, only the adult female takes blood meals, while both sexes take sugar meals. With the recent description of the Anopheles gambiae genome, and with a set of approximately 3000 expressed sequence tags from a salivary gland cDNA library from adult female mosquitoes, we attempted a comprehensive description of the salivary transcriptome of this most important vector of malaria transmission. In addition to many transcripts associated with housekeeping functions, we found an active transposable element, a set of Wolbachia-like proteins, several transcription factors, including Forkhead, Hairy and doublesex, extracellular matrix components and 71 genes coding for putative secreted proteins. Fourteen of these 71 proteins had matching Edman degradation sequences obtained from SDS-PAGE experiments. Overall, 33 transcripts are reported for the first time as coding for salivary proteins. The tissue and sex specificity of these protein-coding transcripts were analyzed by RT-PCR and microarray experiments for insight into their possible function. Notably, two gene products appeared to be differentially spliced in the adult female salivary glands, whereas 13 contigs matched predicted intronic regions and may include additional alternatively spliced transcripts. Most An. gambiae salivary proteins represent novel protein families of unknown function, potentially coding for pharmacologically or microbiologically active substances. Supplemental data to this work can be found at http://www.ncbi.nlm.nih.gov/projects/omes/index.html#Ag2.

Influence of indoor microclimate and diet on survival of Anopheles gambiae s.s. (Diptera: Culicidae) in village house conditions in western Kenya

The survival of female Anopheles gambiae s.s. mosquitoes inside two village house types (grass-thatched and iron-roofed) was studied in relation to diet and ambient indoor microclimatic conditions. Two batches of 20–30, 1-day-old laboratory-bred mosquitoes were maintained inside cages in the grass-thatched (n = 2) and iron-roofed (n = 2) houses and fed daily, one group on 10% glucose and the other on human blood. Throughout the experiments, indoor temperature and relative humidity of the houses were recorded, and mortality of mosquitoes monitored daily until all had died. The experiments were replicated thrice. There was no significant variation in the overall mean temperature (P = 0.93) or relative humidity profiles (P = 0.099) between the two house types, although the iron-roofed houses recorded higher temperature peaks. A Kaplan-Meier survival analysis showed that the mean survival times of mosquitoes were 8 and 10 days in the two grass-thatched huts and 7 and 10 days in the two iron-roof houses for mosquitoes feeding on blood and sugar meals, respectively. The mean survival times of mosquitoes maintained inside similar house types differed only due to diet. In the proportionality of hazards model (Cox regression), the dietary regimes significantly influenced the probability of survival (P = 0.0001), with mosquitoes surviving longer on sugar meals than on blood. Microclimatic factors inside houses also significantly influenced mosquito survival. Although higher peak temperatures were recorded in corrugated iron-roofed houses, the survival of the mosquitoes resting in them did not differ significantly from that in grass-thatched houses. However, the impact of these temperatures on the development of malaria parasites inside the vector needs to be investigated.