Parasitoid wasp uses a venom cocktail injected into the brain to manipulate the behavior and metabolism of its cockroach prey

Unlike other venomous predators, the parasitoid wasp Ampulex compressa incapacitates its prey, the cockroach Periplaneta americana, to provide a fresh food supply for its offspring. We first established that the wasp larval development, from egg laying to pupation, lasts about 8 days during which the cockroach must remain alive but immobile. To this end, the wasp injects a cocktail of neurotoxins to manipulate the behavior of the cockroach. The cocktail is injected directly into the head ganglia using biosensors located on the stinger. The head sting induces first 30 min of intense grooming followed by hypokinesia during which the cockroach is unable to generate an escape response. In addition, stung cockroaches survive longer, lose less water, and consume less oxygen. Dopamine contained in the venom appears to be responsible for inducing grooming behavior. For the hypokinesia, our hypothesis is that the injected venom affects neurons located in the head ganglia, which send descending tonic input to bioaminergic neurons. These, in turn, control the thoracic premotor circuitry for locomotion. We show that the activity of identified octopaminergic neurons from the thoracic ganglia is altered in stung animals. The alteration in the octopaminergic neurons' activity could be one of the mechanisms by which the venom modulates the escape circuit in the cockroach's central nervous system and metabolism in the peripheral system.     

Parasitoid wasp affects metabolism of cockroach host to favor food preservation for its offspring

Unlike predators, which immediately consume their prey, parasitoid wasps incapacitate their prey to provide a food supply for their offspring. We have examined the effects of the venom of the parasitoid wasp Ampulex compressa on the metabolism of its cockroach prey. This wasp stings into the brain of the cockroach causing hypokinesia. We first established that larval development, from egg laying to pupation, lasts about 8 days. During this period, the metabolism of the stung cockroach slows down, as measured by a decrease in oxygen consumption. Similar decreases in oxygen consumption occurred after pharmacologically induced paralysis or after removing descending input from the head ganglia by severing the neck connectives. However, neither of these two groups of cockroaches survived more than six days, while 90% of stung cockroaches survived at least this long. In addition, cockroaches with severed neck connectives lost significantly more body mass, mainly due to dehydration. Hence, the sting of A. compressa not only renders the cockroach prey helplessly submissive, but also changes its metabolism to sustain more nutrients for the developing larva. This metabolic manipulation is subtler than the complete removal of descending input from the head ganglia, since it leaves some physiological processes, such as water retention, intact.     

Wasp manipulates cockroach behavior by injecting venom cocktail into prey central nervous system

The parasitoid wasp Ampulex compressa induces behavioral changes in the cockroach prey by injecting venom into its central nervous system. In contrast to most other venomous predators, the wasp's sting does not induce paralysis. Rather, the two consecutive stings in the thoracic and head ganglia induce three stereotypic behavioral effects. The prey behavior is manipulated in a way beneficial to the wasp and its offspring by providing a living meal for its newborn larva. The first sting in the thorax causes a transient front leg paralysis lasting a few minutes. This paralysis prevents the cockroach from fighting with its front legs, thereby facilitating the second sting in the head. A postsynaptic block of central synaptic transmission mediates this leg paralysis. Following the head sting, dopamine identified in the venom induces 30 minutes of intense grooming that appears to prevent the cockroach from straying until the last and third behavioral effect of hypokinesia commences. In this lethargic state that lasts about three weeks, the cockroach does not respond to various stimuli nor does it initiates movement. However, other specific behaviors of the prey are unaffected. We propose that the venom represses the activity of head ganglia neurons thereby removing the descending excitatory drive to specific thoracic neurons.     

Are monoaminergic systems involved in the lethargy induced by a parasitoid wasp in the cockroach prey?

The venom of the parasitoid wasp Ampulex compressa induces long-lasting hypokinesia in the cockroach prey. Previous work indicates that the venom acts in the subesophageal ganglion to indirectly affect modulation of thoracic circuits for locomotion. However, the target of the venom in the subesophageal ganglion, and the mechanism by which the venom achieves its effects are as yet unknown. While the stung cockroaches appear generally lethargic, not all behaviors were affected, indicating that the venom targets specific motor systems and not behavior in general. Stung cockroaches were observed "freezing" in abnormal positions. Reserpine, which depletes monoamines, mimics the behavioral effects of the venom. We treated cockroaches with antagonists to dopamine and octopamine receptors, and found that the dopamine system is required for normal escape response. Dopamine injection induces prolonged grooming in normal cockroaches, but not in stung, suggesting that the venom is affecting dopamine receptors, or targets downstream of these receptors, in the subesophageal ganglion. This dopamine blocking effect fades slowly over the course of several weeks, similar to the time course of recovery from hypokinesia. The similarity in the time courses suggests that the mechanism underlying the hypokinesia may be the block of the dopamine receptors.     

A Wasp Manipulates Neuronal Activity in the Sub-Esophageal Ganglion to Decrease the Drive for Walking in Its Cockroach Prey

Background

The parasitoid Jewel Wasp hunts cockroaches to serve as a live food supply for its offspring. The wasp stings the cockroach in the head and delivers a cocktail of neurotoxins directly inside the prey''s cerebral ganglia. Although not paralyzed, the stung cockroach becomes a living yet docile ‘zombie’, incapable of self-initiating spontaneous or evoked walking. We show here that such neuro-chemical manipulation can be attributed to decreased neuronal activity in a small region of the cockroach cerebral nervous system, the sub-esophageal ganglion (SEG). A decrease in descending permissive inputs from this ganglion to thoracic central pattern generators decreases the propensity for walking-related behaviors.

Methodology and Principal Findings

We have used behavioral, neuro-pharmacological and electrophysiological methods to show that: (1) Surgically removing the cockroach SEG prior to wasp stinging prolongs the duration of the sting 5-fold, suggesting that the wasp actively targets the SEG during the stinging sequence; (2) injecting a sodium channel blocker, procaine, into the SEG of non-stung cockroaches reversibly decreases spontaneous and evoked walking, suggesting that the SEG plays an important role in the up-regulation of locomotion; (3) artificial focal injection of crude milked venom into the SEG of non-stung cockroaches decreases spontaneous and evoked walking, as seen with naturally-stung cockroaches; and (4) spontaneous and evoked neuronal spiking activity in the SEG, recorded with an extracellular bipolar microelectrode, is markedly decreased in stung cockroaches versus non-stung controls.

Conclusions and Significance

We have identified the neuronal substrate responsible for the venom-induced manipulation of the cockroach''s drive for walking. Our data strongly support previous findings suggesting a critical and permissive role for the SEG in the regulation of locomotion in insects. By injecting a venom cocktail directly into the SEG, the parasitoid Jewel Wasp selectively manipulates the cockroach''s motivation to initiate walking without interfering with other non-related behaviors.     

Wasp uses venom cocktail to manipulate the behavior of its cockroach prey

The sting of the parasitoid wasp Ampulex compressa is unusual, as it induces a transient paralysis of the front legs followed by grooming behavior and then by a long-term hypokinesia of its cockroach prey. Because the wasp's goal is to provide a living meal for its newborn larva, the behavioral changes in the prey are brought about by manipulating the host behavior in a way beneficial to the wasp and its offspring. To this end, the wasp injects its venom cocktail with two consecutive stings directly into the host's central nervous system. The first sting in the thorax causes a transient front leg paralysis lasting a few minutes. This paralysis is due to the presence of a venom component that induces a postsynaptic block of central cholinergic synaptic transmission. Following the head sting, dopamine identified in the venom appears to induce 30 min of intense grooming. During the long-term hypokinesia that follows the grooming, specific behaviors of the prey are inhibited while others are unaffected. We propose that the venom represses the activity of head ganglia neurons thereby removing the descending excitatory drive to the thoracic neurons.Abbreviations CNS central nervous system - DA dopamine - GI giant interneuron - PSP postsynaptic potential - SEG sub-esophageal ganglion - TI thoracic interneuron     

Localization of the site of effect of a wasp's venom in the cockroach escape circuitry

The parasitic wasp Ampulex compressa stings a cockroach Periplaneta americana in the neck, toward the head ganglia (the brain and subesophageal ganglion). In the present study, our aim was to identify the head ganglion that is the target of the venom and the mechanisms by which the venom blocks the thoracic portion of the escape neuronal circuitry. Because the escape responses elicited by a wind stimulus in brainless and sham-operated animals were similar, we propose that the venom effect is on the subesophageal ganglion. Apparently, the subesophageal ganglion modulates the thoracic portion of the escape circuit. Recordings of thoracic interneuron responses to the input from the abdominal giant interneurons showed that the thoracic interneurons receive synaptic drive from these interneurons in control and in stung animals. Unlike normal cockroaches, which use both fast and slow motoneurons for producing rapid escape movements, stung animals activate only the slow motoneuron. However, we show that in stung animals, the fast motoneuron still can be recruited with bath application of pilocarpine, a muscarinic agonist. These results indicate that the descending control from the subesophageal ganglion is presumably exerted on the premotor thoracic interneurons to motoneurons connection of the thoracic escape circuitry. Accepted: 19 December 1998     

Testing adaptive significance of host manipulation with a parasitoid wasp

According to the extended phenotype hypothesis, certain parasites manipulate the host's behaviour which ultimately enhances the transmission of parasite genes into the next generation. The parasitoid wasp Ampulex compressa attacks and stings its cockroach Periplaneta americana host and lays an egg on the cockroach's leg. Before the wasp's oviposition, the stung cockroach engages in excessive self-grooming for about 30 min. The prey location hypothesis posits that self-grooming may allow the wasp to easily locate its host before transporting it to the nest. To test this hypothesis, we manipulated the mobility of the stung cockroach under different spatial constraints. Latencies to locate stung cockroaches with unlimited movement were similar than latencies to locate stung, but motionless cockroaches irrespectively of spatial constraints. Wasps were less willing to lay eggs and seal an entrance to the burrow in treatments with motionless cockroaches which suggests that if the expected reproductive success is low, then parental investment decreases. Our results provide no support for the location hypothesis and call for further experimental investigation of the extended phenotype hypothesis in parasite–host interactions.     

Digger wasp versus cricket: immediate actions of the predator's paralytic venom on the CNS of the prey

The females of the palaearctic digger wasp species Liris niger hunt crickets (e.g., Acheta domesticus) as food for their future brood. The wasps paralyze the prey by injecting their venom directly into each of the three thoracic ganglia and the suboesophageal ganglion. This study describes the effects produced by the Liris venom at the level of the intact prey animal (by chronic electromyogram) and at the level of a dissected preparation (by extra- and intracellular records) during the immediate action. Natural or artificial injections of the Liris venom into various ganglia revealed that: (a) The venom injection induced an about 15- to 35-s long tonical discharge of the neurons located in the stung ganglion. This discharge is usually accompanied by convulsions of the prey's limbs. (b) Subsequently, the generation and propagation of action potentials are blocked for up to 30 min (total paralysis). (c) During total paralysis, the venom blocks synaptic transmission. (d) The effects of the venom are restricted to the stung ganglion. Responses of mechanoreceptors in the legs can be recorded from the peripheral nerves of the stung ganglion during the whole period of total paralysis. (e) The neurons almost completely recover after this period. The venom does not selectively affect leg motoneurons, but affects any neuron (e.g., internerneurons or neurosecretory neurons) in any part of the central nervous system of the prey where it was released.     

Direct injection of venom by a predatory wasp into cockroach brain

In this article, we provide direct evidence for injection of venom by a wasp into the central nervous system of its cockroach prey. Venomous predators use neurotoxins that generally act at the neuromuscular junction, resulting in different types of prey paralysis. The sting of the parasitoid wasp Ampulex compressa is unusual, as it induces grooming behavior, followed by a long-term lethargic state of its insect prey, thus ultimately providing a living meal for the newborn wasp larvae. These behavioral modifications are induced only when a sting is inflicted into the head. These unique effects of the wasp venom on prey behavior suggest that the venom targets the insect's central nervous system. The mechanism by which behavior modifying compounds in the venom transverse the blood-brain barrier to induce these central and long-lasting effects has been the subject of debate. In this article, we demonstrate that the wasp stings directly into the target ganglia in the head of its prey. To prove this assertion, we produced "hot" wasps by injecting them with (14)C radiolabeled amino acids and used a combination of liquid scintillation and light microscopy autoradiography to trace radiolabeled venom in the prey. To our knowledge, this is the first direct evidence documenting targeted delivery of venom by a predator into the brain of its prey.     

Involvement of the opioid system in the hypokinetic state induced in cockroaches by a parasitoid wasp

The parasitoid wasp Ampulex compressa stings and injects venom into the cockroach brain to induce a long-lasting hypokinetic state. This state is characterized by decreased responsiveness to aversive stimuli, suggesting the manipulation of a neuromodulatory system in the cockroach’s central nervous system. A likely candidate is the opioid system, which is known to affect responsiveness to stimuli in insects. To explore this possibility, we injected cockroaches with different opioid receptor agonists or antagonists before they were stung by a wasp and tested the escape behavior of these cockroaches to electric foot shocks. Antagonists significantly decreased the startle threshold in stung individuals, whereas agonists led to an increased startle threshold in controls. Yet, neither agonists nor antagonists had any effect on grooming. To further characterize the interaction between the venom and opioid receptors, we used an antenna-heart preparation. In un-stung individuals external application of crude venom completely inhibits antenna-heart contractions. In stung individuals the antenna-heart showed no contractions. Although acetylcholine restored contractions, the opioid receptor antagonist naloxone was unable to antagonize the venom inhibition. These results suggest that the venom of A. compressa might contribute to the manipulation of cockroach behavior by affecting the opioid system.     

Wasp venom injected into the prey's brain modulates thoracic identified monoaminergic neurons

The wasp Ampulex compressa injects a cocktail of neurotoxins into the brain of its cockroach prey to induce an enduring change in the execution of locomotory behaviors. Our hypothesis is that the venom injected into the brain indirectly alters the activity of monoaminergic neurons, thus changing the levels of monoamines that tune the central synapses of locomotory circuits. The purpose of the present investigation was to establish whether the venom alters the descending control, from the brain, of octopaminergic neurons in the thorax. This question was approached by recording the activity of specific identified octopaminergic neurons after removing the input from the brain or after a wasp sting into the brain. We show that the activity of these neurons is altered in stung and "brainless" animals. The spontaneous firing rate of these neurons in stung and brainless animals is approximately 20% that in control animals. Furthermore, we show that an identified octopamine neuron responds more weakly both to sensory stimuli and to direct injection of current in all treated groups. The alteration in the activity of octopamine neurons is likely to be part of the mechanism by which the wasp induces a change in the behavioral state of its prey and also affects its metabolism by reducing the potent glycolytic activator fructose 2,6-bisphosphate in leg muscle. To our knowledge, this is the first direct evidence of a change in electrical activity of specific monoaminergic neurons that can be so closely associated with a venom-induced change in behavioral state of a prey animal.     

Digger wasp versus cricket: mechanisms underlying the total paralysis caused by the predator's venom

The data presented here describe neurophysiological experiments addressing the question of cellular mechanisms underlying the total paralysis of locomotor behavior in crickets occurring after being stung by females of the digger wasp species Liris niger. The Liris venom effects have been studied by both in vivo recordings from identified neurons of the well-described giant fiber pathway and in vitro recordings from cultured neurons isolated from the terminal ganglion of crickets. The total paralysis of the prey is characterized by a general block of action potential generation as well as by a block of synaptic transmission. Intracellular recordings from neurons in intact ganglia under single electrode voltage-clamp conditions, as well as whole-cell patch-clamp recordings from cultured cricket neurons consistently show that the block of action potential generation by the Liris venom is due to a block of voltage-gated sodium inward currents in neurons of the stung ganglia. Furthermore, our data provide evidence that the Liris venom also blocks calcium currents in identified neurosecretory neurons. On the other hand, outward currents are not affected by the Liris venom. The in vitro recordings suggest that the Liris venom contains active venom components, which, at least for the observed block of inward currents, do not require a metabolic modification. Because venom application does not affect the ACh-induced EPSPs in giant interneurons, the Liris venom does not seem to influence the postsynaptic ACh receptors. The possible pre- and postsynaptic sites of venom action and the functional consequences on synaptic transmission within the giant fiber system are discussed.     

Wasp venom blocks central cholinergic synapses to induce transient paralysis in cockroach prey

The parasitoid wasp Ampulex compressa induces a set of unique behavioral effects upon stinging its prey, the cockroach. It stings into the first thoracic segment inducing 2 to 3 min of transient flaccid paralysis of the front legs. This facilitates a second sting in the cockroach's head that induces 30 min of excessive grooming followed by a 2 to 5-week long lethargic state. In the present study, we examine the immediate effect of the first sting, which is a transient paralysis of the front legs. Using radiolabeled wasps, we demonstrate that the wasp injects its venom directly into the cockroach's first thoracic ganglion. The artificial injection of milked venom into a thoracic ganglion abolishes spontaneous and evoked responses of the motoneurons associated with leg movements. To investigate the physiological mechanism of action of the venom, we injected venom into the last abdominal ganglion of the cockroach, which houses a well-characterized cholinergic synapse. Injected venom abolishes both sensory-evoked and agonist-evoked postsynaptic potentials recorded in the postsynaptic neuron for 2 to 3 min without affecting action potential propagation. Thus, the venom blocking effect has a postsynaptic component that follows the same time course as the transient paralysis induced by the thoracic sting. Finally, injection of a nicotinic antagonist in the front thoracic ganglion induces paralysis of the front legs. We conclude that the transient paralytic effect of the thoracic sting can be mainly accounted for by the presence of a venom active component that induces a postsynaptic block of central cholinergic synaptic transmission.     

Venom spraying inParachartergus colobopterus: A novel defensive behavior in a social wasp (Hymenoptera: Vespidae)

Colonies ofParachartergus colobopterus do not defend against vertebrates by attacking and stinging. Instead, defending workers bend the gaster forward and spray a fine mist of venom in the direction of nearby moving objects. Although venom spraying has been reported forP. fraternus, a species that does sting, we found that this occurred only during sting attempts. We conclude that defensive behavior inP. colobopterus is unique among wasps in that (1) venom is sprayed at intruders by workers standing on the nest and (2) the spray is an atomized mist rather than a stream. We suggest that nest crypticity restricts vertebrate predators on this wasp to small, insect gleaning birds, against which a spray of venom in the eyes, mouth, and nasal passages is more effective than is a stinging defense.     

Smooth muscle contracting compounds in the venom of Megascolia flavifrons (Hym: Scoliidae) with notes on the stinging behaviour

1. The wasp Megascolia flavifrons stings larvae of the stag beetle Oryctes nasicornis on the ventral side of all segments, except the last three, which do not contain nerve ganglia. 2. Experiments indicate a central rather than a peripheral action of the venom. 3. From pharmacological analysis it is concluded that the venom does not contain cholinergic or serotonergic activity, but contains histamine- and bradykinin-like substances. 4. The presence of histamine was confirmed by a radioenzymatic method.     

Venom is beneficial but not essential for development and survival of Nasonia

1. Parasitoid wasps sting and inject venom into arthropod hosts, which alters host metabolism and development while keeping the host alive for several days, presumably to induce benefits for the parasitoid young. 2. This study investigates the consequences of host envenomation on development and fitness of wasp larvae in the ectoparasitoid Nasonia vitripennis, by comparing wasps reared on live unstung, previously stung, and cold‐killed hosts. Developmental arrest and suppression of host response to larvae are major venom effects that occur in both stung and cold‐killed hosts, but not in unstung hosts, whereas cold‐killed hosts lack venom effects that require a living host. Thus, cold‐killed hosts mimic some of the effects of venom, but not others. 3. Eggs placed on live unstung hosts have significantly higher mortality during development; however, successfully developing wasps from these hosts have similar lifetime fecundity to that of wasps from cold‐killed or stung hosts. Therefore, although venom is beneficial, it is not required for wasp survival. 4. While wasps developing on both cold‐killed and stung hosts have similar fitness levels, rearing multiple generations on cold‐killed hosts results in significant fitness reductions of wasps. 5. It is concluded that the largest benefits of venom are induction of host developmental arrest and suppression of host response to larva (e.g. immune responses), although more subtle benefits may accrue across generations or under stressful conditions.     

Absence of chemical alarm in a primitively eusocial wasp (Belonogaster petiolata,Hymenoptera: Vespidae)

Summary The hypothesis thatBelonogaster petiolata (fam. Vespidae) is able to communicate alarm chemically, using odours released with the venom, was tested in bioassays involving presentation of artificial targets to a wasp colony, simultaneously with crushed venom apparatuses. The odour of venom did not lower the threshold of attack and visual stimuli alone (particularly a black, moving object) were sufficient to release attack. Venom odour on a previously stung target probably does not play a role in focusing further attacks on such a target. The results therefore support the null hypothesis that a venom-based alarm pheromone is absent in this species.     

The venom of the wasp Campsomeris sexmaculata (F.) blocks synaptic transmission in insect CNS

1. The action of the venom of the wasp Campsomeris sexmaculata on the insect CNS has been studied using the cercal nerve-giant interneuron preparation of the sixth abdominal ganglion of the cockroach. 2. The venom blocks synaptic transmission either transiently (at low concentration) or for a long time (at higher concentration), and causes a permanent depolarization of the neuron with a delay. 3. The venom does not affect directly the axonal excitability.