Paolo Solari

Gustatory Sensitivity and Food Acceptance in Two Phylogenetically Closely Related Papilionid Species: Papilio hospiton and Papilio machaon

In herbivorous insects, food selection depends on sensitivity to specific chemical stimuli from host-plants as well as to secondary metabolites (bitter) and to sugars (phagostimulatory). Bitter compounds are noxious, unpalatable or both and evoke an aversive feeding response. Instead, sugars and sugar alcohols play a critical role in determining and enhancing the palatability of foods. We assumed that peripheral taste sensitivity may be related to the width of the host selection. Our model consists of two closely phylogenetically related Papilionid species exhibiting a difference in host plant choice: Papilio hospiton and Papilio machaon. The spike activity of the lateral and medial maxillary styloconic taste sensilla was recorded following stimulation with several carbohydrates, nicotine and NaCl, with the aim of characterizing their gustatory receptor neurons and of comparing their response patterns in the light of their different acceptability in feeding behaviour. The results show that: a) each sensillum houses phagostimulant and phagodeterrent cells; b) the spike activity of the gustatory neurons in response to different taste stimuli is higher in P. hospiton than in P. machaon; c) sugar solutions inhibit the spike activity of the deterrent and salt cells, and the suppression is higher in P. machaon than in P. hospiton. In conclusion, we propose that the different balance between the phagostimulant and phagodeterrent inputs from GRNs of maxillary sensilla may contribute in determining the difference in food choice and host range.

Effects of Avermectins on Olfactory Responses of Culicoides imicola (Diptera: Ceratopogonidae)

Abstract The aim of this study was to examine the role of the olfactory system of the midge Culicoides imicola Kieffer as the major system mediating repellency to antihelminthic avermectins. Incidental observations indicate that treatment with Dectomax or Ivomec (commercial formula of the avermectins doramectin and ivermectin, respectively) protects sheep from infection by bluetongue (BT) viruses. Our electrophysiological data from midge antennae showed that the stimulating effectiveness of l-(+)-lactic acid, butanone, and sheep fleece odor decreased after addition of avermectins. The results show that these antihelminthics affect the olfactory sensitivity of the insect toward the animal host by reducing the response to those compounds that attract the insect, consequently reducing the possibility of biting the sheep and thereby transferring the BT virus.

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 hormones 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.

Morpho-functional identification of abdominal olfactory receptors in the midge Culicoides imicola

The aim of the present study was to examine the presence and the possible role of abdominal olfactory sensilla in Culicoides imicola mediating the search for potential hosts and oviposition sites, by means of a morphological, electrophysiological and behavioural approach. The results reported here show that in the midge C. imicola the whole abdomen, comprising the ovipositor, are endowed with three morphotypes of multiporous sensilla that display olfactory sensitivity towards kairomones related to the host-animal skin such as l-(+)-lactic acid and 1-octen-3-ol, to the host-animal urine such as 3-ethylphenol and 4-propylphenol, and to the potent attractant sesame seed oil. Electrophysiological and behavioural data for the first time suggest in the midge the involvement of abdominal olfactory structures in the choice of the oviposition sites and allow in discussing their possible role in the host-animal localisation. Field experiments showed that light traps baited with the aforementioned compounds elicited a stronger degree of attractiveness on midges with respect to the unbaited traps (control), although to a different extent. Our results, while implying a number of considerations concerning the role of molecules tested as kairomones, also suggest their use in the control of the midge C. imicola population.

Transduction mechanism(s) of Na-saccharin in the blowfly Protophormia terraenovae: evidence for potassium and calcium conductance involvement

The study on transduction mechanisms underlying bitter stimuli is a particularly intriguing challenge for taste researchers. The present study investigates, in the labellar chemosensilla of the blowfly Protophormia terraenovae, the transduction mechanism by which saccharin evokes the response of the “deterrent” cell, with particular attention to the contribution of K+ and Ca2+ current and the role of cyclic nucleotides, since second messengers modulate Ca2+, Cl− and K+ currents to different extents. As assessed by extracellular single-sensillum recordings, our results show that the addition of a Ca2+ chelator such as EGTA or the Ca2+ current blockers SK&F-96365, Mibefradil, Nifedipine and W-7 decrease the response of the “deterrent” cell to saccharin. A similar decreasing effect was also obtained following the addition of 4-aminopyridine, a K+ current blocker. On the contrary, the membrane-permeable cyclic nucleotide 8-bromoguanosine 3′,5′-cyclic monophosphate (8Br-cGMP) activates this cell and shows an additive effect when presented mixed with saccharin. Our results are consistent with the hypothesis that in the labellar chemosensilla of the blowfly both Ca2+ and K+ ions are involved in the transduction mechanism of the “deterrent” cell in response to saccharin. Our results also suggest a possible pathway common to saccharin and 8Br-cGMP.

Taste discriminating capability to different bitter compounds by the larval styloconic sensilla in the insect herbivore Papilio hospiton (Géné).

Herbivorous animals may benefit from the capability to discriminate the taste of bitter compounds since plants produce noxious compounds, some of which toxic, while others are only unpalatable. Our goal was to investigate the contribution of the peripheral taste system in the discrimination of different bitter compounds by an herbivorous insect using the larvae of Papilio hospiton Géné as the experimental model, showing a narrow choice range of host plants. The spike activity from the lateral and medial styloconic sensilla, housing two and one bitter-sensitive gustatory receptor neurons (GRNs), respectively, was recorded following stimulation with nicotine, caffeine, salicin and quercitrin and the time course of the discharges was analyzed. Nicotine and caffeine activated all three bitter-sensitive GRNs, while salicin and quercitrin affected only two of them. In feeding behavior bioassays, intact larvae ate glass-fiber disks moistened with salicin and quercitrin, but rejected those with nicotine and caffeine, while lateral sensillum-ablated insects also ate the disks with the two latter compounds. The capability to discriminate bitter taste stimuli and the neural codes involved are discussed.

Stimulation of cyclic AMP formation and nerve electrical activity by octopamine in the terminal abdominal ganglion of the female gypsy moth Lymantria dispar

The biogenic amine octopamine is known to be present in the abdominal ganglia of some insects, but the expression of functional octopamine receptors in these neuronal structures has not yet been characterized. In the present study, we describe the presence in the female gypsy moth terminal abdominal ganglion (TAG), a key structure in the control of the insect reproductive behavior, of an octopamine receptor coupled to stimulation of adenylyl cyclase through the GTP-binding protein G(s). The rank order of potency of different antagonists, which discriminate between the different classes of octopamine receptors, indicated the involvement of the neuronal type 3 receptor. The octopamine-stimulated adenylyl cyclase activity was inhibited by Ca(2+) in the low micromolar range and by activation of either protein kinase A or protein kinase C. In the isolated TAG, bath application of octopamine caused an increase of the spontaneous bursting activity of the emerging nerve of the 5th pair (V), whereas the antagonist mianserin reduced the nerve spiking activity and blocked the stimulatory effect of octopamine. These data demonstrate that the gypsy moth TAG expresses functional octopamine receptors, which may participate in the neuronal control of the insect reproductive behavior.

Chemosensory basis of larval performance of Papilio hospiton on different host plants

Papilio hospiton Géné is an oligophagous species, endemic of the islands of Corsica and Sardinia, using various Apiaceae and Rutaceae as host plants, such as Ferula communis, Ferula arrigonii, Peucedanum paniculatum, Ruta lamarmorae and Pastinaca latifolia. We previously found that the lateral maxillary styloconic sensillum in the larva has two deterrent neurons, one phagostimulant and one salt specific, while the medial sensillum has two phagostimulant neurons, one deterrent and one salt specific. In this work we studied the sensitivity of gustatory receptor neurons (GRNs) to saps of F. communis, F. arrigonii, P. paniculatum, P. latifolia and R. lamarmorae and evaluated the relationship between taste sensitivity to different host-plants and larval growth rate on each of them. The spike activity was recorded from medial and lateral taste sensilla stimulated with plant saps, and GRN response patterns were cross compared in the light of a different feeding acceptance. The phagodeterrent GRNs show a higher activity in response to F. arrigonii and R. lamarmorae than to F. communis, P. paniculatum and P. latifolia. Behavioral trials showed that the time to pupation is significantly longer when larvae are reared on F. arrigonii and R. lamarmorae than on the other host-plants. These results suggest that the different activity of the phagodeterrent GRNs may inhibit food acceptance and extend the duration of the larval stage.

Regulatory mechanisms and the role of calcium and potassium channels controlling supercontractile crop muscles in adult Phormia regina.

Bioassays and electrophysiological recordings were conducted in the adult blowfly Phormia regina to provide new insights into the regulatory mechanisms governing the crop filling and emptying processes of the supercontractile crop muscles. The cibarial pump drives ingestion. Simultaneous multisite extracellular recordings show that crop lobe (P5) distension during ingestion of a 4.7 μl sugar meal does not require muscle activity by any of the other pumps of the system. Conversely, pumping of fluids toward the anterior of the crop system during crop emptying is brought about by active muscle contraction, in the form of a highly coordinated peristaltic wave starting from P5 and progressively propagating to P6, P4 and P3 pumps, with P5 contracting with a frequency about 3.4 times higher than the other pumps. The crop contraction rate is also modulated by hemolymph-borne factors such as sugars, through ligand recognition at a presumptive receptor site rather than by an osmotic effect, as assessed by both behavioural and electrophysiological experiments. In this respect, sugars of equal osmolarity produce different effects, glucose being inhibitory and mannose ineffective for crop muscles, while trehalose enhances crop activity. Finally, voltage and current clamp experiments show that the muscle action potentials (mAPs) at the P4 pump are sustained by a serotonin-sensitive calcium conductance. Serotonin enhances calcium entry into the muscle cells and this could lead, as an indirect modulatory effect, to activation of a Ca(2+)-activated K(+) conductance (IK(Ca)), which sustains the following mAP repolarization phase in such a way that further mAPs can be generated early and the frequency consequently increased.

Antennular Morphology and Contribution of Aesthetascs in the Detection of Food-related Compounds in the Shrimp Palaemon adspersus Rathke, 1837 (Decapoda: Palaemonidae)

Shrimp are an essential ecological component of marine ecosystems, and have commercial importance for human consumption and aquaculture. Like other decapod crustaceans, shrimp rely on chemical senses to detect and localize food resources by means of chemosensilla that are located mainly on the cephalothoracic appendages. Using the shrimp Palaemon adspersus, a model organism with omnivorous feeding behavior, we aimed to provide comparative information on the role of aesthetascs, antennular sensilla, and flicking behavior in food detection. To this end, we examined i) the morphology of antennular sensilla by field emission scanning electron microscopy, ii) the shrimp’s sensitivity to a number of food-related compounds (amino acids and sugars) by means of whole-animal bioassays, and iii) the contribution of the aesthetasc sensilla to food detection. Our results showed that, aside from the aesthetascs, only three other main morphotypes of setae with chemoreceptive features were present in the antennules, thus accounting for relatively simple sensillar equipment. Nevertheless, we found broad-spectrum sensitivity of the shrimp to a number of amino acids (i.e., isoleucine, leucine, methionine, phenylalanine, glycine, tryptophan, cysteine, and tyrosine) and carbohydrates (trehalose, maltose, cellobiose, and fructose) that was consistent with the omnivorous or scavenging habits of the animal. Although aesthetasc ablation attenuated flicking behavior in a chemical stimulus-independent manner, success in detection and short-range localization of food did not rely on the presence of aesthetasc sensilla. This finding confirms the existence of a non-aesthetasc alternative pathway for feeding, with functional redundancy in simple generalist feeder models such as shrimp.