Olfactory Associative Learning of the Pupal Parasitoid Pimpla luctuosa Smith (Hymenoptera: Ichneumonidae)

Laboratory experiments were conducted to determine the role of learning in olfactory host searching by the ichneumonid pupal parasitoid, Pimpla luctuosa Smith. Females learned to associate novel odors such as vanilla and strawberry with hosts when they oviposited in at least several hosts with the odors. Repeated experiences of hosts with an odor increased the response of the experienced odor, and females that had experienced host odor seven times responded to the experienced odors 90% of the time. Although the response by females to a learned odor gradually decreased with increasing host-deprivation time, 60% of the females that had experienced host odor 7 days earlier still responded to the experienced odor. Females also learned two separate odors associated with hosts at a time and responded to both odors without a preference for one odor over the other. When trained two separate odors with hosts, females learned the second odor more quickly than the first odor. After females experienced several stings in simulated hosts with the previously learned odor, they ceased to respond to the learned odor, suggesting that repeated unrewarding experiences cause females to cease to respond to the learned odors.     

Effects of Wind Speed and Atmospheric Pressure on Mate Searching Behavior in the Aphid Parasitoid Aphidius nigripes (Hymenoptera: Aphidiidae)

The effects of wind speed and atmospheric pressure on male mate searching behavior, modulated by a female sex pheromone, were investigated in the aphid parasitoid Aphidius nigripes (Hymenoptera: Aphidiidae). Male A. nigripe generally did not reach females at wind speeds of 100 cm/sec, as the majority of individuals taking flight in the pheromone plume (81.8%) were unable to sustain upwind flight. At lower wind velocities, male responsiveness to females generally decreased with distance from the source. However, wind speeds approaching the upper threshold (100 cm/sec) tended to eliminate this distance effect. Therefore, there appears to be a trade-off between the need for higher wind speeds to detect the pheromone source from long distances, and a reduction in male flight capacity as wind velocity increases. Our results also indicate that chemical communication in A. nigripes could be affected by variations in atmospheric pressure, as we observed a relationship between pressure fluctuations in the 24 hr prior to testing and male responsiveness to females. The importance of these abiotic factors on mate searching behavior is discussed within the context of the reproductive biology of A. nigripes.     

The Influence of Oviposition Experience on Response to Host Pheromone in Trichogramma sibericum (Hymenoptera: Trichogrammatidae)

Searching times and residence times of Trichogramma sibericum Sorokina were measured in the laboratory on individual cranberry leaves that had been treated with the sex pheromone of blackheaded fireworm [Rhopobota naevana (Hübner)] and on leaves that were left untreated. Mean searching time was higher on leaves treated by passive diffusion with either 50 or 100 g of the main component of fireworm pheromone, (Z)-11-tetradecen-1-ol acetate (Z11-14: Ac), than on control leaves. Mean residence times were also higher on leaves treated by passive diffusion with 50 g of Z11-14: Ac than on leaves untreated with pheromone. Pretrial oviposition experience in either the presence or the absence of host pheromone did not influence variation in searching time or residence time. This indicates that neither associative learning of the odor of host pheromone nor a more generalized increase in response to chemical stimuli after oviposition (priming) affects retention responses of T. sibericum to pheromone. Results are discussed in the context of current theories on the evolution of learning in insect parasitoids and as they relate to the concurrent use of pheromone-based mating disruption and releases of T. sibericum for pest management of the blackheaded fireworm.     

Reproductive Biology ofCephalonomia hyalinipennis(Hymenoptera: Bethylidae), a Native Parasitoid of the Coffee Berry Borer,Hypothenemus hampei(Coleoptera: Scolytidae), in Chiapas, Mexico

The coffee berry borer,Hypothenemus hampei(Ferrari) (Coleoptera: Scolytidae), is the most important pest of coffee worldwide.Cephalonomia hyalinipennisAshmead (Hymenoptera: Bethylidae) was recently discovered naturally attackingH. hampeiin Mexico.C. hyalinipennisboth preys upon and parasitizesH. hampei.We report laboratory investigations on the reproductive biology ofC. hyalinipennisaimed at assessing the suitability of this parasitoid for culturing and release in biological control programs. Adult females lived up to 95 days (mean for mated females = 57 days), and risk of death increased with age. Mating status, reproductive effort, and female size influenced adult female longevity, but only weakly. The number ofH. hampeipreyed upon did not affect longevity. Rates of attack (predation plus parasitism) declined with age, but the proportion of attackedH. hampeithat were parasitized increased. Overall, about half of the attackedH. hampeiwere preyed upon and half were parasitized. Females received approximately 46 sperm per mating and 77% became sperm depleted before the end of their reproductive lives. Estimated mean lifetime fecundity was 88 eggs. Offspring survival from egg to adulthood was 60%, with batches of eggs tending to survive or die collectively; 21.2% of surviving progeny were males. These results suggest thatC. hyalinipennismay be suitable for mass rearing and release, but further work is needed to investigate interactions betweenC. hyalinipennisand other parasitoids ofH. hampei.     

Sex Allocation by a Parasitoid Wasp (Hymenoptera: Ichneumonidae) to Different Host Species: A Question for the Mechanism of Host Size Estimation

Sex allocation by the polyphagous solitary pupal parasitoid wasp Pimpla luctuosa Smith to a small host species, Galleria mellonella (L.), and a large host species, Mamestra brassicae L., was investigated to test whether female wasps responded to hosts of different sizes across different host species. In the experiments, both host species were presented to each test female wasp. Primary and secondary sex ratio experiments revealed that female wasps laid more female eggs in larger pupae of each host species, indicating that female wasps recognized size differences within host species. The wasp sex ratio (male ratio) from M. brassicae, however, was much higher than that expected on the basis of the sex ratio curve from different-sized G. mellonella. Larger hosts of each host species yielded larger wasps, indicating that the host size estimation by female wasps across different host species was incomplete or was not simple. These results suggested that P. luctuosa evaluated host size not only by physical measures such as dimension but also by other unknown measures. A possible explanation for the adaptiveness of different sex ratio responses by Pimpla luctuosa to different host species was discussed.     

Influence of physiological state and experience on the responsiveness ofLysiphlebus testaceipes (Cresson) (hymenoptera: aphidiidae) to aphid honeydew and to host plants

The effect of physiological state (hunger) and experience on the responsiveness of the aphid parasitoid, Lysiphlebus testaceipes(Cresson), to clean and honeydew-contaminated host plants was investigated in laboratory bioassays. Both fed and unfed parasitoids spent significantly longer examining honeydew-contaminated plants compared to uncontaminated controls, but the presence of honeydew did not influence attack latency (i.e., the speed with which naive parasitoids found and attacked hosts). Hunger, however, had a significant negative effect on attack latency, presumably as a result of a physiologically based shift from host- to food-location behavior in starved parasitoids. The parasitoid 's response to clean plants was significantly increased as a result of classical conditioning procedures, whereas the response to honeydew-contaminated plants was not.     

Exploitation of the Fecal Shield of the Lily Leaf Beetle, Lilioceris lilii (Coleoptera: Chrysomelidae), by the Specialist Parasitoid Lemophagus pulcher (Hymenoptera: Ichneumonidae)

We investigated signal sources used by the parasitoid Lemophagus pulcher in locating and accepting larvae of its host, the lily leaf beetle, Lilioceris lilii. Olfactometer bioassays revealed that larvae with fecal shields, larvae without shields, the shield alone, and lily leaves damaged by L. lilii were all attractive to female parasitoids. In contact bioassays, L. pulcher females were attracted to shields and showed ovipositor probing independently of whether the larva underneath was L. lilii or a nonhost, suggesting that the shield plays a primary role in short-range host location and host acceptance by L. pulcher. The attractiveness of the shield is at least partly of a chemical nature, since shield extracts applied to dummies increased contact duration and induced ovipositor probing by L. pulcher. Another putative defense system of L. lilii, i.e., oral discharge which is emitted by disturbed larvae, was also attractive to experienced, but not to naive, female parasitoids. In all other tests, naive and experienced female L. pulcher responded to the same signal sources tested, suggesting that the host-selection behavior of this biological control candidate is governed largely by innate responses to host-associated cues.     

Associative Learning in Response to Color in the Parasitoid Wasp Nasonia vitripennis (Hymenoptera: Pteromalidae)

A parasitoid that can learn cues associated with the host microenvironment should have an increased chance of future host location and thereby increase its reproductive success. This study examines associative learning in response to simultaneous exposure to the colors yellow and blue in mated females of the parasitoid wasp Nasonia vitripennis. Preference was measured as the proportion of time spent on a color. When trained with one color rewarded with hosts and honey and the other unrewarded, females showed an increase in preference for the rewarded color with increasing number of training days (1, 3, and 7 days). Hosts and honey together produced a slightly greater preference toward the rewarded color than just hosts, which produced a greater preference than just honey. When trained with a variable reward on one color and a constant reward on the other, females preferred the color associated with the variable reward when it was yellow, but not when it was blue. Thus, relative to no reward, the presence of a variable reward decreased the strength of preference toward the constantly rewarded color. Finally, females trained with regular hosts on one color and used hosts on the other preferred the color associated with the regular hosts when that color was blue but showed no preference in the reverse situation. The presence of used hosts instead of no reward did not increase the strength of preference for the color associated with the regular hosts.     

Comparison of Host-Searching and Ovipositional Behavior of Cardiochiles nigriceps Viereck (Hymenoptera: Braconidae), a Parasitoid of Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae), in Tobacco and Cotton

To compare host-searching and ovipositional behavior of Cardiochiles nigriceps Viereck (CN) in tobacco and cotton, behavior of CN females was observed on Hellothis virescens (Febricius)-infested plants and recorded using a Psion Organizer II with Observer software. Hover, search, oviposit, preen, rest, walk, agony-search, drop-search, and miss-search were the nine behaviors observed for each plant species. Agony-search was a searching behavior exhibited when CN females were prevented from ovipositing in a host because they were unable to reach the host. Drop-search and miss-search were the searching behaviors that occurred by CN females after a host dropped from the plant and when CN females missed ovipositing in a host, respectively. Hovering, searching, oviposition, miss-search, resting, and walking were equally common for CN on tobacco and cotton. Agony-search was exhibited more on tobacco than cotton, while drop-search and preening occurred more on cotton than tobacco. Female CN spent more time hovering in tobacco than in cotton, indicating that these females preferred tobacco to cotton. Female parasitoids also spent more time in the main ovipositional sequence, hover–search–oviposit, in tobacco than in cotton, mainly because females hovered more in tobacco than in cotton. Frequency and time of behavioral sequences associated with experiencing frustration in ovipositing due to difficulty in reaching or finding a host and subsequent level of success in ovipositing in a new host were determined. Overall, the rate of successful oviposition by CN females was significantly higher in tobacco (87.7%) than in cotton (79.5%). For cotton and tobacco, the rate of successful oviposition by CN females was significantly reduced (ca. 40.0%) when they experienced frustration in ovipositing compared to the rate of ovipositional success (100%) for CN females when hosts were present and easily accessible. Encountering ovipositional frustration also increased the duration of time for successful oviposition in comparison to the time spent for problem-free oviposition in cotton and tobacco. Interestingly, with successful oviposition, the amount of time spent in a behavioral sequence was not significantly different for tobacco and cotton regardless of whether females experienced ovipositional frustration or not. However, female CN invested more time in tobacco than in cotton when they were unsuccessful in ovipositing due to the inaccessibility or disappearance of hosts. CN females' preference for tobacco over cotton probably gave them more motivation to continue searching for hosts, especially those not easily attained, on tobacco than on cotton. Nevertheless, CN females readily searched in cotton for HV even in the presence of host-infested tabacco. CN females possibly could be successful in maintaining levels of HV below an economic threshold when using a tobacco trap crop to protect cotton as the main crop.     

Volatile and Contact Chemicals Released by Nezara viridula (Heteroptera:Pentatomidae) Have a Kairomonal Effect on the Egg Parasitoid Trissolcus basalis (Hymenoptera: Scelionidae)

The responses of females of the egg parasitoid Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae) to volatile and contact chemicals from its host Nezara viridula (L.) (Heteroptera: Pentatomidae) were investigated in a Y-tube olfactometer and under open arena conditions. In the Y-tube tests, volatiles from virgin males and from females in a preovipositional state attracted T. basalis females, while volatiles from host virgin females did not. In an open arena, traces left by N. viridula adults in different physiological conditions function as contact cues inducing the wasps to remain longer in the arena and to change the pattern of their walking behavior. However, only contact kairomones from N. viridula mated females in a preovipositional condition induced an arrestment response characterized by an increase in patch searching time and turning rates and a reduction in linear speed. The chemical ecological implications of these results on this host–parasitoid association are discussed.     

Host Age Response in the Parasitoid Wasp Spalangia cameroni (Hymenoptera: Pteromalidae)

Female Spalangia cameroni produced more offspring from younger house fly pupae, both when given a choice of host ages and when not given a choice. Host age did not affect offspring survivorship. Offspring were larger when they had developed on younger hosts and the effect was independent of offspring sex. Having previously parasitized old hosts versus young hosts did not reduce a female's production of offspring in subsequent hosts. Females distinguished between young and old hosts both in the light and in the dark. Females did not distinguish between host ages prior to physical contact with the host but could distinguish by the time they first began exploring a host by tapping it with their antennae; thus, they could distinguish before drilling into a host.     

Effect of Temperature on the Life History of Cirrospilus sp. near lyncus (Hymenoptera: Eulophidae), a Parasitoid of Phyllocnistis citrella (Lepidoptera: Gracillariidae)

The effect of four constant temperatures on the life history of Cirrospilus sp. near lyncus was examined in the laboratory. This species is one of the most abundant generalist indigenous parasitoids of the citrus leafminer, Phyllocnistis citrella Stainton, in Spain. Adult lifespan of C. sp. near lyncus decreased from 50.2 to 9.1 days as temperatures increased from 15 to 30°C, respectively. Both gross fecundity and host-feeding were highest at 20°C (170.48 eggs and 32.33 hosts). Oviposition rates were optimal at higher temperatures (5.22 eggs per day at 25°C and 4.79 eggs per day at 30°C) and were dependent on female age. In contrast, host-feeding rates for a given temperature did not depend on age. Generation time decreased with increasing temperatures from 68.05 days at 15°C to 12.19 days at 30°C. Net reproduction peaked at 20°C (68.86 viable females per female). Intrinsic rate of increase doubled from 15°C (0.059 females per female per day) to 20°C (0.127 females per female per day) and almost doubled again from 20 to 30°C (0.210 females per female per day). Given these parameters, C. sp. near lyncus could perform optimally in the area occupied by P. citrella in the Mediterranean region.     

Foraging Behaviour and Patch Exploitation by Campoplex dubitator (Hymenoptera: Ichneumonidae), a Parasitoid of Bark-mining Larvae

This study examines the foraging behaviour of Campoplex dubitator Horstmann (Hymenoptera: Ichneumonidae), a parasitoid of the highly concealed bark-mining host, Enarmonia formosana Scopoli (Lepidoptera: Tortricidae), and how this foraging behaviour is affected by host density. The ultimate aim was to determine whether foraging behaviour and patch fidelity could explain patterns of parasitism in the field. An ethogram was first constructed to portray C. dubitator behaviour prior to and immediately following host attacks. The pre-attack behavioural sequence was highly structured whereas behavioural transitions became less predictable after a sting event. Females spent more time on patches with higher host densities and host encounters caused a significant reduction in the leaving tendency. The giving up time was only slightly affected by host encounters. Campoplex dubitator did not demonstrate the ability to distinguish between occupied and empty host mines. The results show that C. dubitator forages more efficiently at lower host densities and this behavioural phenomenon may result in the patterns of parasitism observed in the field.     

Odor learning byLeptopilina boulardi,a specialist parasitoid (Hymenoptera: Eucoilidae)

We studied odor learning in Leptopilina boulardi,a specialist larval parasitoid of Drosophila melanogaster.The behavioral responses of differently experienced females to an artificial odor (Must de Cartier, Paris) were analyzed using a fourarmed airflow olfactometer. The responses of females with an oviposition experience in the presence of the perfume were compared with those of four control groups. As controls we used naive females, females with an oviposition experience in the absence of odor, females which had been previously exposed to perfume but without an oviposition experience, and females with an oviposition experience which also had been exposed to perfume but not at the same time. The results demonstrate that a specialist such as L. boulardican learn very well to respond to an artificial odor by associating this odor with a reward, i.e., an oviposition. The four control groups responded more or less in a similar way.     

Cloning of cDNA for Vitellogenin of the Parasitoid Wasp, Pimpla nipponica (Hymenoptera: Apocrita: Ichneumonidae): Vitellogenin Primary Structure and Evolutionary Considerations

The cDNA for vitellogenin (Vg) of the parasitoid wasp Pimpla nipponica (Hymenoptera: Apocrita) was cloned and sequenced.¹ The deduced amino acid sequence with 1807 residues was obtained. The N-terminal 20 amino acids chemically determined for vitellin (Vn) agreed completely with the deduced 20 amino acids that follow the 16 amino acid residues for putative signal peptide. The cDNA clone for the Vg of the turnip sawfly Athalia rosae (Hymenoptera: Symphyta), previously obtained and partially sequenced, was also completely sequenced and the amino acid sequence deduced. Amino acid sequences were compared between these two species and also with known Vg sequences from other insects. Common to all these insects is the presence of two long regions with relatively well-conserved amino acid sequences, one near the N-terminal extending 267–282 residues (including two cysteines at conserved locations), and the other starting at position 450 to 655 and extending 279–283 residues, and of a region at the C-terminal extending some 200 residues (about 250 in Aedes aegypti due to the presence of a serine-rich stretch) with 10 cysteines at conserved locations. A molecular phylogenetic tree was constructed.     

Influence of adult experience on host microhabitat location by the generalist parasitoid,Campoletis sonorensis (Hymenoptera: Ichneumonidae)

The effect of adult experience on microhabitat location behavior of the generalist parasitoid Campoletis sonorensis (Cameron)was examined in a wind tunnel bioassay. Responses were tested to the odors of two host plants (cotton and sesame) of Heliothis virescens (F.) or a nonhost plant (potato), either damaged and infested with host larvae and host products (host/plant complex) or intact, clean and uninfested. Parasitoid females remained naive or were allowed one oviposition experience on either of the plants, 1 min, 2 h, or 24 h prior their introduction into the wind tunnel. In a no-choice test, parasitoids experienced 1 min prior to bioassay completed significantly more flights to sesame and potato host/plant complexes than did naive parasitoids. However, 24 h after experience, only females experienced on potato completed more flights to the host/ plant complex than did naive females. Parasitoids experienced 1 min prior to flight to undamaged plants showed a slight increase in flight response (significant only for potato) but, after 24 h, completed only as many flights as naive parasitoids. In a dual-choice situation, parasitoids did not show a preference for either of the two host plants but did prefer a host to a nonhost plant. This innate plant preference was not changed by a single oviposition experience. The potential significance of these results to the microhabitat location behavior of C. sonorensisin the field is discussed.     

In Vitro Rearing of the Parasitoid Exorista larvarum (L.) (Diptera: Tachinidae) on Meat Homogenate-Based Diets

The tachinid Exorista larvarum (L.), a polyphagous gregarious larval endoparasitoid of Lepidoptera, was reared from egg to fecund adult on media containing commercial meat homogenates for babies as the main ingredient. Four media, each containing a diverse homogenate supplemented with extract of Galleria mellonella L. pupae, were tested first. Despite the difference in nutrient content, the kind of homogenate did not significantly affect the adult yields (30.2 to 40.7%) or puparial weights. Two other diets free of host materials (I and II) were then tested. Both were based on Gerber veal homogenate combined with different amounts of yeast extract and chicken egg yolk and were supplemented with wheat germ (I) or saccharose (II). Adult yields (28.7 to 32.7%) and puparial weights did not differ significantly between the two diets. Fly longevity and fecundity of the females obtained on diet I were comparable to those of the females emerged from puparia formed in G. mellonella larvae. Male and female puparial weights were, however, higher and development times longer on the diet than in the host.     

Field Evaluation of Amitus bennetti (Hymenoptera: Platygasteridae), a Parasitoid of Bemisia argentifolii (Hemiptera: Aleyrodidae), in Cotton and Bean

Field cage experiments were conducted in Riverside, California to quantify the impact of releases of the parasitoid Amitus bennetti Viggiani & Evans on mortality of the whitefly Bemisia argentifolii Bellows & Perring. Single-row 50-m-long plots were planted with either cotton or bean. Cages were erected over the plants in each row, and adult whiteflies were released into the cages. Approximately 10 days later, adult parasitoids were released. Marked individual whiteflies were scored every 4 days for 6 weeks. Paired life tables were then constructed from census data from release and control cages over a single whitefly generation. Total whitefly mortality in release cages (71% in bean, 61% in cotton) was significantly greater than in control cages (25% in bean, 34% in cotton). The marginal rate for mortality attributable directly to the parasitism was 0.535 in the bean plots and 0.201 in the cotton plots. In addition, other mortality was greater in the release plots, possibly reflecting death of parasitized hosts before larval parasitoids could complete development. Parasitism was the greatest mortality factor in the study.