Physiology of feeding preference patterns of female black blowflies (Phormia regina Meigen)

Summary The relative effectiveness of 0. 1 M sucrose and 10% yeast extract in eliciting feeding by femalePhormia regina has been shown to be related to the fly's carbohydrate reserve levels. Females that have had ad lib. access to 0.1 M sucrose (S flies, standard condition) take larger drinks of 10% yeast than 0.1 M sucrose starting at three days of age, indicating that 10% is the more effective feeding stimulus. Females that have had ad lib. access to 0.1 M sucrose and 10% yeast (S/Y flies, standard condition) always drink more 0.1 M sucrose than 10% yeast (Fig. 1). S/Y (standard condition) flies appear to respond to the yeast as if it were dilute carbohydrate: the volumes of 0.1 M sucrose and 10% yeast that they drink fluctuate in parallel. S (standard condition) flies respond to the yeast differently: starting day 3, their drink volumes of 10% yeast increase as drink volumes of 0.1 M sucrose decrease.The elevation of the volume of 0.1 M sucrose drunk by flies whose carbohydrate reserve levels have been depleted by diet, starvation or flight (Figs. 2–4) supports the hypothesis that some physiological state or process related to carbohydrate metabolism regulates the fly's responsiveness to yeast extract. Possible mechanism by which the fly translates carbohydrate reserve level into feeding preference are discussed. Available evidence points to a CNS phenomenon, probably involving the stomatogastric nervous system. The pattern of the selective feeding cycle of femalePhormia can be explained in terms of the accumulation and utilization of carbohydrate reserves.This work was supported by a Spencer Foundation Predoctoral Fellowship from Princeton University and by NSF Grant number BNS 76-02172 to Dr. Vincent G. Dethier. I am grateful to the Departments of Zoology and Entomology of the University of Massachusetts, Amherst, for the use of their facilities. Many thanks to Drs. V.G. Dethier, Elizabeth Bowdan and David Falk for critical discussion and assistance.     

The diverticulated crop of adult Phormia regina

The crop of adult Phormia regina consists of a duct that diverges from the esophagus, just in front of the cardia, and extends ventrally and posteriorly into the thorax and abdomen where it forms a bilobed sac. Flattened epithelial cells produce the cuticular lining of the crop. When empty, or partially full, the epithelial cells and cuticular lining form folds extending into the lumen, thus providing for expansion as the crop sac fills. Covering the sac on the hemolymph side is a layer of anastomosed, intrinsic muscles connected to one another by intercellular cytoplasmic bridges. Mitochondria are located at the periphery of the sarcomere. Also inside the sarcomere are glycogen, sarcoplasmic reticula, and transverse tubular systems (T-system). I, A, and Z-bands are present and the Z-bands are not in register making the muscle-type supercontractile. Important structures, not previously researched and associated with the crop muscles, are the crop nerves. Coming off the corpora cardiaca, and running down each side of the crop duct, is a pair of nerves, each housing several axons. These nerves extend to and branch over the crop sac. Here they penetrate the muscle mass and form neuromuscular junctions where electron-dense droplets of neurosecretion are released. Based on the literature, and research in our laboratory, it has now been shown that these nerves carry adipokinetic hormone, Drosophila insulin-like peptide, and a dromyosuppressin-like neuropeptide.     

The effect of diet on activity in male Calliphora vicina blowflies

ABSTRACT. The effect of diet on the activity of male Calliphora vicina R.D. (Diptera, Calliphoridae) was studied. Protein–fed males (PF) were significantly more active than males fed only on sugar and water (NPF); as a consequence they ingested more sugar. The phenomenon was probably due to PF insects spending more time in flight than NPF, as no significant differences were found between the wingbeat frequencies and flight speeds of tethered flies of the two treatments. Protein ingestion also had a physiological effect, as PF flies had heavier thoraces than NPF flies with increased titres of protein.     

Central control of feeding in the diapausing adult blowfly Phormia regina.

1. The tarsal acceptance threshold of non-diapausing adult blowflies rises with age. 2. Diapausing flies have a non-significantly elevated tarsal acceptance threshold, but drink significantly less sucrose solution than their non-diapausing counterparts. 3. Removal of diapausing flies to non-diapause inducing conditions resulted in a lowering of the threshold. 4. Failure to demonstrate a significant difference between the input signal from the peripheral receptors of non-diapausing and diapausing flies showed that the inhibition of feeding in diapausing flies was not due to peripheral inhibition but was controlled centrally.     

Presence and regulation of trehalose in the body of adult Phormia regina

Trehalose, the major blood sugar of Phormia regina, is present within its tissues in an amount exceeding that in the total blood volume. A major part of the reserve is found in the abdominal fat body. An investigation of trehalose regulation, pursued with the use of a trehalose tolerance test, indicates that within a period of 4 h the adult fly can remove from its blood amounts of this sugar in excess of twice its normal level. The surplus is dealt with in an as yet unknown way, being either sequestered in the tissues (not as trehalose or glucose), metabolized, or excreted in a form other than trehalose or glucose. The process is regulated by the head, and a link between the body and the head must be maintained throughout the entire period of activity.     

The effects of Calliphora vomitoria Tachykinin-I and the FMRFamide-related peptide Perisulfakinin on female Phormia regina crop contractions, in vitro

The dipteran crop is an elaborate diverticulation of the foregut that serves as an important food reservoir and feeding regulator. Peptidergic innervation has been associated with the crop of the blow fly Phormia regina and myotropic neuropeptides have been previously demonstrated to affect crop contraction rates, in vitro. The blow fly peptide, callitachykinin-1 was found to increase the rate of contractions and alter the contractile morphology of the P. regina crop, in vitro. The cockroach peptide perisulfakinin, however, had no measurable affect on crop contractions.     

Regulation of mating behavior by nutrition and the corpus allatum in both male and female Phormia regina (Meigen)

Both dietary protein and the corpus allatum (CA) were required for normal mating behavior in both male and female black blow fly, Phormia regina (Meigen). Nutrition (protein diet) activated the CA in both sexes. More than 10 mg of dietary liver was required for each male to result in 80% insemination of females, while 20 mg of liver was required for each female to allow 78% of females to become inseminated. Between 10 to 15 mg of protein meal (i.e., liver) was required to activate sexual receptivity in 71% of the females, while between 15 to 20 mg of liver was needed to support full o?cyte development in 70% of the females (Yin, C.-M., Zou, B.-X., Li, M.-F., Stoffolano, J.G., Jr. 1994. Discovery of a midgut peptide hormone which activates the endocrine cascade leading to oogenesis in Phormia regina Meigen. Journal of Insect Physiology, 40, 283-292). Allatectomy suppressed mating behavior more than 2-fold in both sexes. Topical application of 10 &mgr;g of S-methoprene (a juvenile hormone analogue) at 12 h after the onset of liver feeding restored sexual activity of both allatectomized males and females. Incidence of successful insemination increased as the o?cyte development progressed. Ovariectomy suppressed sexual receptivity more than 3-fold in liver-fed females. Thus, in addition to nutrition and the CA, ovaries and their developmentaal status can also affect the sexual receptivity in female P. regina.     

Effect of alkaloids on feeding by Phormia regina

Eight plant-derived alkaloids are shown to significantly reduce intake of sugar solutions by adult Phormia regina Meigen. The alkaloids differed in degree of inhibition caused, and their relative effectiveness as deterrents was similar with 2, 4, and 6 day old flies. Results are discussed with reference to current ideas on possible modes of action of plant secondary compounds acting as feeding deterrents to phytophagous insects.

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Résumé Huit alcaloïdes réduisent la consommation de solutions sucrées chez les adultes de Phormia regina Meigen. Ces alcaloïdes diffèrent selon le degré d'inhibition induit. Leur efficacité relative en tant que substances dissuadantes est la même pour les mouches agées de 2, 4 ou 6 jours. La spartéine, l'atropine, la tomatine, la quinine et la papavérine sont les plus efficaces, provoquant une baisse de l'alimentation de plus que 80% en 6 h. L'étude démontre clairment que les alcaloödes végétaux influencent l'alimentation de cette mouche polyphage. Les auteurs discutent des implications de ces résulats sur notre perception des effects des composés végétaux secondaires sur les systèmes sensoriels des insects phytophages.

     

Conditioning odor-shock associations in the black blowfly,Phormia regina

Discriminative classical conditioning of an olfactory avoidance response was demonstrated in the blowfly, Phormia regina.Learning indices were calculated as the fraction of flies avoiding the CS+ (conditioned stimulus paired with electric shock reinforcement) minus the fraction of flies avoiding the CS-(conditioned stimulus not paired with electric shock), averaged over two different groups of flies, in which reciprocal odors were used as the CS+. Avoidance responses to both odors presented simultaneously at a T-maze choice point yielded mean learning indices of zero for naive flies or pseudoconditioned (shock alone) or sensitized (odors alone) controls. In contrast, pairing an odor with electric shock produced a mean learning index significantly greater than zero. These results are similar to those reported for Drosophila melanogaster.Blow flies, however, showed much lower levels of associative learning than fruit flies.     

Allatotropic activity in the brain of female Phormia regina (Diptera: Calliphoridae)

In Phormia regina, the rate of juvenile hormone (JH) synthesis rises rapidly after the ingestion of an adequate protein meal. In a previous publication we have localized the neurons containing Manduca sexta allatotropin (Mas-AT)-like substances in the brain of P. regina and demonstrated the allatotropic effect of synthetic Mas-AT in sugar-fed flies in vitro. In this current study, we examined the possible role of the brain of P. regina after the fly received a protein meal. In vitro studies showed that the brain releases, at 8 h after a protein meal, a factor(s) with a strong allatotropic effect. This factor(s) stimulates the corpus allatum (CA) to produce 6.9 times more juvenile hormones (JHs) than the control CA. Time course studies showed that the release of this allatotropic factor(s) is temporally controlled. Only the brains collected from flies at 6 and 8 h after the onset of a liver meal release allatotropic factor(s). Injection of anti-Mas-AT antiserum partially suppressed the fly follicle development in vivo. Presence of anti-Mas-AT antiserum decreased the allatotropic effect of the brain released allatotropic factor(s) in vitro. In addition to a Mas-AT-like substance, it is possible that the brains of liver-fed P. regina females may synthesize other allatotropic factors that are chemically unrelated or partially related to Mas-AT, which cannot be recognized/neutralized by our anti-Mas-AT antiserum.     

The effects of injection of amphetamine on female insemination in the black blow fly, Phormia regina (Diptera: Calliphoridae)

Abstract. An earlier study on the blowfly Phormia regina (Meigen) demonstrated that the injection of amphetamine (12 μg) depletes biogenic amine levels in the CNS. In the present study, P. regina females were injected with amphetamine (12 μg), each female was placed with three males and insemination success was determined. Amphetamine inhibited female insemination by 43.3% at 2–90 min post-injection and by 70% at 10–60 min post-injection. At 180–270 min post-injection, there was no significant inhibition of female insemination. This study indicates a possible role in insects for the biogenic amines in female insemination.     

Role of Phote-HrTH (Phormia terraenovae hypertrehalosemic hormone) in modulating the supercontractile muscles of the crop of adult Phormia regina Meigen

Phote-HrTH (Phormia terraenovae hypertrehalosemic hormone) has been demonstrated in the Diptera to be involved in flight metabolism, reproduction, and diapause. Each of these events needs the hormone’s action and requirement for carbohydrates is the common denominator. In Diptera, carbohydrates are taken up during feeding by action of the cibarial pump and are then stored in the crop. Using adult Phormia regina, both a bioassay and electrophysiological recordings show that Phote-HrTH slows down or inhibits the crop lobe muscles (P5) and, at the same time, stimulates the muscles of the pump 4 (P4) involved in pushing fluids out of the crop and up into the midgut for digestion. The EC₅₀ for P4 was in the nanomolar range while the IC₅₀ for P5 was 1.4–75.1 pM. The effect of Phote-HrTH on P4/5 suggests that the peptide is important in coordinating the two pumps, which are involved in moving carbohydrates up into the midgut for digestion. The adult crop organ is an essential storage organ for carbohydrates and now should be considered an important structure capable of delivering nutrients to the midgut for digestion.     

The role of olfaction in the stimulation of oviposition in the blowfly,Phormia regina

The odour of a liquid medium containing powdered milk and yeast stimulates Phormia regina to oviposit. In the presence of a sufficiently high odour concentration, contact stimulation by the same medium does not increase egg laying. The receptors chiefly involved in the reception of the odour are located on the antennae and/or palps. Evidence is given for olfactory reception by the ovipositor.     

Faeces feeding by adult Phormia regina (Diptera: Calliphoridae): impact on reproduction

resh beef liver, sugar, and five different types of faeces were evaluated as supportive diets for egg development in the blowfly Phormia regina . Females on a sugar diet were unable to develop follicles beyond stage 3, whereas liver proved to be the best diet for complete egg maturation. Some faecal diets were unable to support egg maturation when fed upon for a short period of time; however, longer periods of feeding produced complete egg maturation. The necessity to feed for longer periods of time in order to produce eggs on most of these diets was attributed to their low protein content. Males, in a shorter period of time than females, obtained enough protein from faeces to activate the neuroendocrine system involved in mating.     

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

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