Feeding behaviour of the horsefly Tabanus nigrovittatus (Diptera: Tabanidae): effects of dissolved solids on ingestion and destination of sucrose or ATP diets

Abstract Wild caught female horseflies, Tabanus nigrovittatus Macq. (Diptera: Tabanidae), were fed solutions containing either sucrose or ATP as a phagostimulant. The sucrose diets were presented as free liquids at room temperature. The ATP in the other diets was dissolved in 0.15 M NaCl, or in 0.3 M or 1 M cellobiose and was presented covered with a Parafilm membrane at 38^oC. The content of dissolved solids in some diets was adjusted with added cellobiose, which had no phago-stimulatory effect. Diets of 1 M or 0.6 M sucrose were ingested in large amounts and were directed to the crop. Diets of 0.3 M or 0.1 M sucrose induced lower levels of ingestion, and the diet was directed to the midgut. The addition of cellobiose to give a 1 M total concentration of dissolved solids in these sucrose diets induced many insects responding to the lower concentrations of sucrose to direct the diet to the crop. Diets of ATP in saline or in 0.3 M cellobiose were directed to the midgut. ATP in 1 M cellobiose caused many of the insects to put the diet in both the crop and the midgut. Two behavioural models are presented that integrate interactions between the level of phagostimulation, the concentration of dissolved solids, and an endogenous feeding factor. The dissolved solids effect is most likely a response to osmotic pressure.     

Effect of plasma components on the feeding response of the mosquito Aedes aegypti L. to adenine nucleotides

ABSTRACT. The gorging response of Aedes aegypti to the ATP dissolved in platelet-poor plasma is greater than that of ATP dissolved in 0.15 m NaCl. The plasma components NaHCO₃ and albumin account for the full effect of the potentiation. Phosphate or tris buffers do not duplicate the bicarbonate effect. In 0.15 m NaCl with bicarbonate but lacking albumin the concentrations inducing 50% feeding are 58 μM ATP, 140 μM ADP, 460 μM AMP and 1500 μM cAMP. Non-adenine nucleotides such as ITP and GTP and phytic acid, and 2,3-diphosphoglyceric acid had no activity.     

Innate immune response of Aedes aegypti

Insects are able to protect themselves from invasion by pathogens by a rapid and potent arsenal of inducible immune peptides. This fast, extremely effective response is part of the innate immunity exhibited by all insects and many invertebrates, and shows striking similarities with the innate immune response of vertebrates. In Aedes aegypti invasion of the hemocoel by bacteria elicits the production of defensins, cecropins, a peptide active only against Gram-negative bacteria, and several other peptides that we are now characterizing. However, not all insects utilize the same peptides in the same concentrations, which may reflect the pathogens to which they may have been exposed through evolutionary time. These protective measures we see in mosquitoes are the current state of the evolution of a rapid immune response that has contributed to the success of insects in inhabiting essentially every niche on earth. The molecules involved in the response of Aedes aegypti to pathogens, and the potential role of these peptides against eukaryotic parasites ingested and transmitted by mosquitoes are discussed.     

Feeding responses of the horsefly, Tabanus nigrovittatus, to phagostimulants

ABSTRACT. Wild caught horseflies, Tabanus nigrovittatus Macq. (Diptera, Tabanidae), were presented solutions of 0.15 MNaCl at 37°C containing various concentrations of ATP, ADP, AMP, adenosine, phytic acid or 2,3-diphosphoglycerate in an artificial feeding apparatus. The insects fed upward through a Para-film M® membrane. ADP (ED₅₀ 35 μM) was more potent than ATP (ED₅₀ 112 μM) and AMP (ED₅₀ 382 mUM). All of these diets were deposited in the midgut, an indication that the flies were in the 'blood feeding' mode. Adenosine caused only 23% gorging at 1 mM. Phytic acid caused only 10% gorging at 1 mM and 2,3-diphosphoglyceric acid had no activity at 0.6 mM. Flies would feed only in highly reflective cages under high levels of light intensity (1200–1500 lux) at the membrane surface.     

Ingestion and diet destination in Culiseta inornata: responses to water, sucrose and cellobiose

ABSTRACT. At ambient temperature, contact with free liquids containing sugar induces Culiseta inornata Williston (Diptera: Culicidae) females to adopt the nectar-feeding mode of behaviour. While in this mode, two distinct control mechanisms appear to operate: one controlling the amount of ingestion, the other diet destination. Mosquitoes were exposed to water and various concentrations of sucrose and cellobiose in water. Only small or taste amounts of water were imbibed, and it was directed to the midgut. Cellobiose solutions of less than 0.1M were treated as water. Cellobiose was phagostimulatory; as the concentration increased from 0.1 to 0.5 M the percentage of insects ingesting large amounts rose from 11% to 68%. Cellobiose diets were directed primarily to the midgut. Sucrose was also phagostimulatory. As the concentration increased from 0.025 to 0.2 M, the percentage of insects ingesting large amounts increased from 20 to 92. At lower sucrose levels, insects ingesting large amounts put it mainly into the crop, while insects ingesting smaller amounts put the diet into the midgut or both the crop and midgut. At sucrose concentrations of 0.4 M or more, most insects took large amounts and directed it to the crop. Different combinations of sucrose and cellobiose with a total concentration of 0.5 M all caused insects to take mainly large meals. At sucrose concentrations of 0.2M and above, the insects responded as they would to sucrose alone, directing the diet to the crop. At sucrose concentrations of 0.1M or less, diet destination patterns were more complex, with individual animals depositing the diet in the crop, the midgut, or in both. A behavioural model is presented which postulates different receptors for sucrose and cellobiose, control of diet destination via differential control of midgut and crop valves, and variation of threshold values according to endogenous factors.     

Sex-specific differences in nutrient regulation in a capital breeding caterpillar, Spodoptera litura (Fabricius)

Nutrient requirements by male and female insects are likely to differ, but relatively little is known regarding how sexes differ in their regulation of macronutrient acquisition. The present study reports the results from a laboratory experiment in which behavioural and physiological components of nutrient regulation were compared between male and female caterpillars of Spodoptera litura (Fabricius). When provided with choices between two nutritionally complementary foods (one is a protein-biased diet and the other a carbohydrate-biased diet), both males and females adjusted their food selection to defend an intake target. However, the composition of diet preferred by the two differed, with females selecting significantly more protein than males with no difference in carbohydrate intake between the two. When confined to single diets with varying mixtures of protein and carbohydrate [P:C ratios, expressed as the percentage of diet by dry mass: protein 42%:carbohydrate 0% (p42:c0), p35:c7, p28:c14, p21:c21, p14:c28, p7:c35], females consumed more macronutrients than did males across on all P:C diets except the extremely carbohydrate-biased diet (p7:c35). Under both choice and no-choice feeding condition, such sex differences in nutrient intake were not expressed until late in the feeding stage of the final stadium. Sexes also differed in post-ingestive utilization of ingested nutrients. Females utilized ingested protein for body growth with greater efficiency compared to males, presumably reflecting provisioning their adult needs for protein to develop eggs, whereas males were more efficient at depositing lipids from carbohydrate intake than females.     

Effects of temperature and diet on development and interspecies competition in Aedes aegypti and Aedes albopictus

We asked whether climate change might affect the geographic distributions of Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae). We tested the effects of temperature, diet and the presence of congeneric species on the performance of immature stages of these two aedine species in the laboratory. Mosquitoes in three different species-density combinations were reared at four constant temperatures (20 °C, 25 °C, 30 °C, 35 °C) on low- or high-level diets. Of the four temperatures tested, mortality increased only at 35 °C in both species. Mortality was higher on the high-level diet than on the low-level diet at 35 °C, but not at other temperatures. The presence of congeneric species had a significant positive effect on mortality in Ae. albopictus, but not in Ae. aegypti. Both species developed more quickly at higher temperatures within the range of 20-30 °C; development was not enhanced at 35 °C. Population growth of Ae. albopictus was more stable, regardless of diet and temperature; that of Ae. aegypti varied more according to these two factors. These species-specific attributes may help to explain the latitudinal distribution of the mosquitoes and degree of species dominance where they are sympatric.     

Interactions of dietary protein and carbohydrate determine blood sugar level and regulate nutrient selection in the insect Manduca sexta L

The non-homeostatic regulation of blood sugar concentration in the insect Manduca sexta L. was affected by nutritional status. Larvae maintained on diets lacking sucrose displayed low concentrations of trehalose, the blood sugar of insects, which varied from 5 to 15 mM with increasing dietary casein level between 12.5 and 75 g/l. These insects were glucogenic, as demonstrated by the selective 13C enrichment of trehalose synthesized from [3-13C]alanine, and de novo synthesis was the sole source of blood sugar. The distribution of 13C in glutamine established that following transamination of the 13C substituted substrate, [3-13C]pyruvate carboxylation rather than decarboxylation was the principal pathway of Pyr metabolism. The mean blood trehalose level was higher in insects maintained on diets with sucrose. At the lowest dietary casein level blood trehalose was approximately 50 mM, and declined to 20 mM at the highest casein level. Gluconeogenesis was detected in insects maintained on sucrose-free diets at the higher protein levels examined, but [3-13C]pyruvate decarboxylation and TCA cycle metabolism was the principal fate of [3-13C]alanine following transamination, and dietary carbohydrate was the principal source of glucose for trehalose synthesis. Feeding studies established a relationship between nutritional status, blood sugar level and dietary self-selection. Insects preconditioned by feeding on diets without sucrose had low blood sugar levels regardless of dietary casein level, and when subsequently given a choice between a sucrose diet or a casein diet, selected the former. Larvae preconditioned on a diet containing sucrose and the lowest level of casein had high blood sugar levels and subsequently selected the casein diet. Larvae maintained on the sucrose diet with the highest casein level had low blood sugar and self-selected the sucrose diet. When preconditioned on diets with sucrose and intermediate levels of casein, insects selected more equally between the sucrose and the casein diets. It is concluded that blood sugar level may be intimately involved in dietary self-selection by M. sexta larvae, and that in the absence of dietary carbohydrate, gluconeogenesis provides sufficient blood sugar to ensure that larvae choose a diet or diets that produce an optimal intake of dietary protein and carbohydrate.     

Proteolysis in the gut of mosquito larvae results in further activation of the Bacillus sphaericus toxin

Gut proteases from the larvae of the mosquito Culex pipiens convert the 43-kilodalton (kDa) toxin from Bacillus sphaericus 2362 to a 40-kDa peptide. The 50% lethal concentration of this peptide for tissue culture-grown cells of Culex quinquefasciatus was 1.0 microgram/ml (as determined by the intracellular ATP assay), 54-fold less than that of the 43-kDa peptide. Gut proteases from Anopheles gambiae and Aedes aegypti, as well as bovine pancreatic trypsin, also converted the 43-kDa protein to a 40-kDa peptide which was indistinguishable from the peptide formed by the proteases from C. pipiens with respect to its toxicity to tissue culture-grown cells of C. quinquefasciatus. Evidence for the in vivo conversion of the 43-kDa protein to the 40-kDa peptide was also obtained from experiments in which larvae of C. pipiens, Anopheles gambiae, and Aedes aegypti were fed crystals from B. sphaericus 2362. By using the exclusion of trypan blue as an indication of cell viability, it was shown that chitobiose, chitotriose, N-acetylmuramic acid, and N-acetylneuraminic acid decreased the toxicity of the 40-kDa peptide (from 100 to 50% mortality at about 10 mM concentrations of these sugars). Muramic acid, N-acetylgalactosamine, and N-acetylglucosamine were less effective, while several sugars had no effect, suggesting that the 40-kDa toxin binds to specific receptors on the cell membrane. The 40-kDa protein was less toxic to tissue culture-grown cells of Anopheles gambiae and Aedes dorsalis, and the same sugars which reduced the toxicity for cells of C. quinquefasciatus were also effective in reduction of toxicity for these cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)     

Proteolysis in the gut of mosquito larvae results in further activation of the Bacillus sphaericus toxin.

Gut proteases from the larvae of the mosquito Culex pipiens convert the 43-kilodalton (kDa) toxin from Bacillus sphaericus 2362 to a 40-kDa peptide. The 50% lethal concentration of this peptide for tissue culture-grown cells of Culex quinquefasciatus was 1.0 microgram/ml (as determined by the intracellular ATP assay), 54-fold less than that of the 43-kDa peptide. Gut proteases from Anopheles gambiae and Aedes aegypti, as well as bovine pancreatic trypsin, also converted the 43-kDa protein to a 40-kDa peptide which was indistinguishable from the peptide formed by the proteases from C. pipiens with respect to its toxicity to tissue culture-grown cells of C. quinquefasciatus. Evidence for the in vivo conversion of the 43-kDa protein to the 40-kDa peptide was also obtained from experiments in which larvae of C. pipiens, Anopheles gambiae, and Aedes aegypti were fed crystals from B. sphaericus 2362. By using the exclusion of trypan blue as an indication of cell viability, it was shown that chitobiose, chitotriose, N-acetylmuramic acid, and N-acetylneuraminic acid decreased the toxicity of the 40-kDa peptide (from 100 to 50% mortality at about 10 mM concentrations of these sugars). Muramic acid, N-acetylgalactosamine, and N-acetylglucosamine were less effective, while several sugars had no effect, suggesting that the 40-kDa toxin binds to specific receptors on the cell membrane. The 40-kDa protein was less toxic to tissue culture-grown cells of Anopheles gambiae and Aedes dorsalis, and the same sugars which reduced the toxicity for cells of C. quinquefasciatus were also effective in reduction of toxicity for these cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effects of artificial diets on the anterior intestinal cell ultrastructure of Rhodnius prolixus (Hemiptera:Reduviidae)

Mated female Rhodnius prolixus were fed diets of washed rabbit erythrocytes (RBC), rabbit plasma, edestin (a plant storage protein) in Ringer's or Ringer's solution alone. All diets contained 1 mM ATP. The effects of these diets on anterior intestinal cell ultrastructure were evaluated and compared to changes induced by normal blood feeding. Separation of basal labyrinth membranes was induced earlier after Ringer's-feeding than with all other diets. Normal modifications of the rough endoplasmic reticulum (rer) were observed only in response to blood components, while rer cisternae were swollen shortly after edestin feeding. Normal modifications to lysosomal structure were induced by plasma and RBC feeding, but changes were also observed with the non-blood diets. Production of extracellular membrane layers (ECML) by the microvilli occurred in response to each diet: plasma-feeding induced considerably early ECML formation, but prolonged ECML proliferation was greatest after RBC feeding. Storage inclusions were most prominent in cells of plasma-fed insects. Initial changes in the basal labyrinth may be induced by a high luminal fluid: protein ratio; in the rer by blood factor(s); and in lysosomes by plasma factor(s). Gut distention initiates ECML production but continued proliferation is regulated by the midgut contents.     

Effects of azadirachtin on oogenesis in Aedes aegypti

ABSTRACT. . Azadirachtin in blood fed to adult female Aedes aegypti through an artificial membrane does not cause feeding inhibition over a wide dose range (0–200 ng/female), and high doses of ingested azadirachtin fail to inhibit or delay oviposition. However, significant, transient retardation of oocyte growth is observed for up to 72 h after feeding. Immature oocytes are observed in 86% of azadirachtin-fed females decapitated 10 h after a blood meal, whereas 96% of decapitated control females contain maturing oocytes. This suggests that azadirachtin delays the release of one or more factors from the head that regulate oogenesis. We propose that adult females overcome the effect of azadirachtin by rapid metabolism rather than by excretion of the compound, since by 2 h after a blood meal, only 0.1% of ingested azadirachtin was recovered from excreta and 5% recovered from the body.     

Molecular analysis of the Aedes aegypti carboxypeptidase gene family

To gain a better understanding of coordinate regulation of protease gene expression in the mosquito midgut, we undertook a comprehensive molecular study of digestive carboxypeptidases in Aedes aegypti. Through a combination of cDNA cloning using degenerate PCR primers, and database mining of the recently completed A. aegypti genome, we cloned and characterized 18 A. aegypti carboxypeptidase genes. Bioinformatic analysis revealed that 11 of these genes belong to the carboxypeptidase A family (AaCPA-I through AaCPA-XI), and seven to the carboxypeptidase B gene family (AaCPB-I through AaCPB-VII). Phylogenetic analysis of 32 mosquito carboxypeptidases from five different species indicated that most of the sequence divergence in the carboxypeptidase gene family occurred prior to the separation of Aedes and Anopheles mosquito lineages. Unlike the CPA genes that are scattered throughout the A. aegypti genome, six of seven CPB genes were found to be located within a single 120 kb genome contig, suggesting that they most likely arose from multiple gene duplication events. Quantitative expression analysis revealed that 11 of the A. aegypti carboxypeptidase genes were induced up to 40-fold in the midgut in response to blood meal feeding, with peak expression times ranging from 3 to 36 h post-feeding depending on the gene.     

Bloodmeal microfilariae density and the uptake and establishment of Wuchereria bancrofti infections in Culex quinquefasciatus and Aedes aegypti.

The relationship between ingestion of microfilariae (mf), production of infective larvae (L3) and mf density in human blood has been suggested as an important determinant in the transmission dynamics of lymphatic filariasis. Here we assess the role of these factors in determining the competence of a natural vector Culex quinquefasciatus and a non vector Aedes aegypti to transmit Wuchereria bancrofti. Mosquitoes were infected via a membrane feeding procedure. Both mosquito species ingested more than the expected number of microfilariae (concentrating factor was 1.28 and 1.81 for Cx. quinquefasciatus and Ae. aegypti, respectively) but Cx. quinquefasciatus ingested around twice as many mf as Ae. aegypti because its larger blood meal size. Ae. aegypti showed a faster mf migration capacity compared to Cx. quinquefasciatus but did not allow parasite maturation under our experimental conditions. Similar proportions of melanized parasites were observed in Ae. aegypti (2. 4%) and Cx. quinquefasciatus (2.1%). However, no relationship between rate of infection and melanization was observed. We conclude that in these conditions physiological factors governing parasite development in the thorax may be more important in limiting vectorial competence than the density of mf ingested.     

Compensation by locusts for changes in dietary nutrients: behavioural mechanisms

ABSTRACT. The detailed behavioural mechanisms underlying an instance of compensation for changes in dietary nutrients are described for the first time in an insect. Nymphs of Locusta migratoria L. were given one of four artificial diets on the third day of the fifth instar, and their feeding patterns recorded in detail for 12 h. The diets represented combinations of two protein and two digestible carbohydrate levels (28% and 14% dry weight) presented in an otherwise complete nutrient mix. At the nutrient levels used, locusts regulated their intake of food with respect to protein but not digestible carbohydrate in the diet. They ate more of the lower protein diets by eating the same sized meals more frequently than insects fed on the higher protein diets. Compensation was not complete over the 12 h observation period: insects on the lower-protein diets ingested and absorbed 72% as much nitrogen as those insects fed on the higher-protein diets. Possible physiological mechanisms underlying the behavioural mechanisms are discussed.     

Plant feeding site selection on soybean by the facultatively phytophagous predator Orius insidiosus

Experiments were conducted to test whether the facultatively phytophagous predator Orius insidiosus (Say) (Heteroptera: Anthocoridae) ingested phloem, xylem or mesophyll contents from soybean plants (Glycine max L.). Potential uptake of phloem sap was examined by radiolabeling photosynthate with ¹⁴CO₂ and then measuring the accumulation of radiolabeled metabolites in feeding animals. Most O. insidiosus feeding on radiolabeled plants ingested no or very low levels of label; only 3% ingested small amounts of label, indicating the experimental insects fed very little, if at all, on the phloem. In contrast, well known phloem feeding insects used as positive controls accumulated substantial levels of labeled metabolites after feeding on known host plants. O. insidiosus did feed on xylem contents, as shown by ingestion of safranin-labeled xylem fluid. A few of the insects showed signs of feeding on the mesophyll, as indicated by the presence of chloroplasts in the gut. However, the small diameter of the food canal may cause limited passage of chloroplasts, which would contribute to an underestimation of the frequency of mesophyll feeding. Some radiolabeled metabolites remain in the mesophyll so those insects that ingested low levels of radiolabel probably ingested label from the mesophyll, which supports the notion that some level of mesophyll feeding occurred. Feeding site determines the nutrients ingested during phytophagy. These insects obtain water from the xylem, and may ingest small amounts of starches, sugars, and amino acids from the mesophyll. The results suggest that facultative phytophagy by this heteropteran predator primarily provides the insect with water, but also may provide some nutrients that supplement a prey diet and help the predator survive periods when prey are scarce.     

Effect of ATP analogues on the gorging response of Aedes aegypti

ABSTRACT. The ATP analogues adenylylimidodiphosphate and adenylylmethylenediphosphate are 3–5-fold more effective than ATP as gorging stimulants for Aedes aegypti. This increased potency is not due to the fact that the two analogues are not hydrolysed by the mosquito salivary apyrase, but most likely to their greater affinity to the mosquito gustatory receptor protein. The analogues 2'd ATP and 3'd ATP are about half as potent as ATP, while 2',3'-dideoxyadenosine triphosphate is 10-fold more potent than ATP in evoking the gorging response. It is proposed that removal of both hydroxyl groups eliminates binding of the stimulant at the ribose moiety, thus allowing the molecule greater freedom to rotate and bind more effectively to its two other binding sites at the amino group on the purine and at the terminal phosphate. Our data demonstrate that ATP activates the gorging response of Ae.aegypti merely by binding to its receptor protein and is not required as an exogenous source of energy. Gorging response to ATP is competitively inhibited by novobiocin.     

Consumption of a toxic food by caterpillars increases with dietary exposure: support for a role of induced detoxification enzymes

We examined whether there is a link between feeding behavior and detoxification enzyme activity, which increases over time in proportion to the ingested dose of toxic allelochemicals. We hypothesized that, for insects consuming a toxic but nondeterrent food, consumption would initially decline but then increase after continuous exposure in conjunction with the induction of detoxification enzyme activity. We tested this hypothesis by observing the feeding behavior of fall armyworms (Spodoptera frugiperda) when offered diets containing indole 3-carbinol (I3C), a well-known inducer of detoxification enzymes in this species. The I3C concentrations used were toxic but nondeterrent, thereby avoiding any confounding influence of preingestive responses. The feeding pattern of caterpillars previously unexposed to dietary I3C (i.e. with basal detoxification enzyme activity) was substantially altered when offered I3C-containing diets. Compared with control caterpillars fed I3C-free diets, they exhibited fewer feeding bouts and had a greater proportion of long (> 60 min) non-feeding pauses between bouts. In contrast, the feeding pattern of caterpillars pre-exposed to an I3C diet (i.e. with induced detoxification enzyme activity) did not differ from that of control caterpillars. We conclude that there is a relationship between feeding behavior and detoxification activity in these caterpillars, and propose two postingestive mechanisms that could link these processes.     

Pre-ingestive and post-ingestive effects of soya bean extracts and rutin on Trichoplusia ni growth

Total ethanolic crude extracts from soya bean (Glycine max L. Merrill) PI 227687 and `Davis', three purified fractions (A, B and C) from PI 227687, and pure rutin were tested for their effect on the growth of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) larvae, from egg hatching to pupation. Analysis of covariance (ANCOVA), followed by bicoordinate utilisation plots, was used to remove the effect of feeding time from consumption and weight of pupae and to separate pre- ingestive and post- ingestive effects of different diets on T. ni growth. Survival, pupal weight, and feeding time of T. ni were all negatively affected by PI 227687 total extracts. Pupal weight and consumption remained affected by diet, after removing the effect of feeding time by ANCOVA. Larvae fed on the `Davis' extract diet grew larger and consumed more rapidly a larger amount of food than those fed on PI 227687 and control diets. T. ni pupal weight was strongly influenced by consumption. In general, those insects that consumed more food were heavier but larvae fed on diet containing PI 227687 extract grew less than the others, irrespective of the amount of ingested and digested food. The feeding time was adversely affected by PI 227687 fraction A, composed of two flavonol glycosides (rutin and quercetin 3-O-glucosylgalactoside) and of an isoflavone glycoside (genistin). Survival and pupal weight were not affected, although the consumption and assimilation of ingested food was lower when larvae were fed on fractions A and C. The toxicity of the higher rutin concentration (2%) was similar to that observed when the insects were fed on unfractioned PI 227687, except that effects on food consumption were not observed. Results indicated that the feeding deterrence of PI 227687 must be caused by other compounds from fractions A and C. Moreover, rutin, acting as an antibiotic, appears to account for the adverse effects of PI 227687 on the physiology of T. ni.     

Structural organization of posterior midgut muscles in mosquitoes, Aedes aegypti and Anopheles gambiae

In order to adapt to diverse feeding behavior, animal intestines have evolved with distinct differences. Such adaptation may include the structure of the longitudinal and circular muscles that maintain the integrity and the tensile strength of the gut tissue in higher metazoans. Here we examined the structural organization of the posterior midgut muscles of two insects, Aedes aegypti and Anopheles gambiae. We found the estimated number of longitudinal muscles in a cross-section to be 168 in Ae. aegypti and 37 in An. gambiae. Within the region, the estimated number of circular muscles is 77 in Ae. aegypti and 57 in An. gambiae. In An. gambiae, longitudinal muscles appear as sets of parallel bundles. Each set overlaps its neighbor to form a continuous tube. We found that this novel mode of muscle fiber sharing makes all circular muscles interconnected. Both types of muscle lie orthogonally to form a grid that holds the epithelium of the posterior midgut. In Ae. aegypti, the muscle fibers between the bundles are shared extensively, making the organization more intricate. This study implies that, because of its simple structure, the insect midgut may provide a powerful tool with which to study the structural evolution and function of animal intestines.