Quality and environmental factors affecting Trichogramma brassicae efficiency under field conditions

Field parasitism of the egg parasitoid Trichogramma brassicae Bezdenko (Hymenoptera, Trichogrammatidae) (synonymous to T. maidis Pint. et Voegele) on Ostrinia nubilalis Hübner was compared to four single quality parameters (walking speed, fecundity on the factitious host Ephestia keuhniella Zeller, fecundity on the natural host O. nubilalis, and life span) previously measured in the laboratory and, a quality index calculated from three of the measured parameters. A single quality parameter (fecundity on E. keuhniella) and the calculated quality index showed a correlation to T. brassicae field parasitism, for different T. brassicae populations. The number of female Trichogramma released clearly influenced parasitism but not in the same proportion for all populations tested, reflecting that numbers released can only to some extent compensate for low quality in Trichogramma. Variation indegree hours above 18°C also influenced field parasitism. When incorporating degree hours to the quality index a significant correlation R²=0.56 (P=0.01) is obtained. In this work fecundity of T. brassicae on E. keuhniella can be as good an indicator for the potential field performance of T. brassicae, as the quality index which takes into account three quality parameters. Since environmental factors can obscure the potential performance of a population, i.e., one which under optimal laboratory conditions performs well, the quality index gives only a partial indication of how the released strains will perform in the field.     

Locomotor activity ofTrichogramma brassicae (Hymenoptera) under the influence of high-temperature shocks

In order to assess the effects of high temperatures prevailing during the releases ofTrichogramma brassicae Bezdenko parasitoid used against the European corn borerOstrinia nubilalis Hübner, we exposed wasps in the white pupa stage (WP) to single or double shocks (at 35 or 44°C for 6 h) in the laboratory. The locomotor activity of adults in the presence ofO. nubilalis egg masses is greater than in pure air. Single or double shocks at 35°C have little effect on the females. On the contrary, wasps exposed to single or double shocks at 44°C had a lower motility than control wasps.     

Mise en évidence d'une activité kairomonale des œufs de la pyrale du maïs sur les femelles deTrichogramma maidis

Résumé Des extraits de la surface d'œufs de Pyrale du ma?s ont été testés en microenceinte et dans un olfactomètre linéaire sur des ♀♀ deTrichogramma maidis. En microenceinte, les extraits stimulent la locomotion et le comportement exploratoire des ♀♀. En olfactomètre ils stimulent à distance la remontée des tubes vers la source. Ces résultats impliquent la présence de substances à activité kairomonale sur les œufs de Pyrale du ma?s. Des analyses préliminaires des extraits ont été faites par chromatographie en phase gazeuse couplée ou non à la spectrométrie de masse et démontrent la présence de nombreux hydrocarbures.

---

Summary Extracts from the surface of eggs of the European corn borer were tested in small observation chambers and in a linear olfactometer for their effects on the searching behaviour of the egg-parasitoidTrichogramma maidis. In the observation chambers, the extracts elicited exploratory behaviour by female parasitoids with no previous oviposition experiences and increased the time the females spent in walking and drumming their antennae. Recording the position of 10 ♀♀ each minute for 15 min in 2 tubes of an olfactometer, indicated that upwind motion was significantly greater in the tube containing 10 μl of an egg extract than in the control tube containing just solvent. It is concluded that kairomones are present onO. nubilalis eggs and could act as searching incitants and short-range attractants. Preliminary GC and GC/MS analysis revealed the abundance of hydrocarbons in the extracts but the active chemicals were not characterized.

     

Does mass rearing of field collected Trichogramma brassicae wasps influence acceptance of European corn borer eggs?

The purpose of our study was to identify a trait that changes quickly during Trichogramma mass rearing, and that could therefore be used to monitor stock deterioration. Quality deterioration in mass reared Trichogramma wasps was evaluated by examining host acceptance behaviour of Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) on the target host Ostrinia nubilalis Hübner (Lepidoptera: Crambidae). We compared three replicate lines (designated ‘E’) reared in the laboratory on the factitious host Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) for 27 generations, a line ‘O’ reared in the laboratory on O. nubilalis for 24 generations, and a line ‘F’ reared in the laboratory on O. nubilalis for only two generations. All lines were initiated from field collections of O. nubilalis egg masses. We also evaluated natal host effects by rearing each line on E. kuehniella as well as on O. nubilalis for the last generation prior to testing. The percentage of wasps accepting the O. nubilalis egg mass was significantly higher for the E lines (69.6%) than for the F line (46.5%), while wasps of the O line showed intermediate (57.4%) acceptance. Thus, wasps laboratory reared on E. kuehniella performed better than wasps which had recently been collected in the field. Wasps of the O line showed extended probing behaviour compared to the other lines. Lines did not differ in the duration from the first host contact to the beginning of the drilling, probing or trembling behaviour. Natal host (E. kuehniella or O. nubilalis) did not affect acceptance of the target host O. nubilalis. Even though there is some evidence of adaptation to laboratory rearing conditions, we found no indication for quality deterioration in terms of acceptance behaviour of the target host O. nubilalis when T. brassicae was mass reared on the factitious host E. kuehniella.     

Release density,efficiency and disappearance ofTrichogramma nubilale for control of European corn borer

We evaluated how manyTrichogramma nubilale should be released at a single location to controlOstrinia nubilalis in sweet corn. Six 8.6×16 m plots received 18.4 to 2 090 ΦΦT. nubilale/SAI when plants were in the mid to late whorl stage, where SAI, surface area index, is the plant surface area/m². To evaluate the potential control by our releases, we exposed laboratory-rearedO. nubilalis egg masses to the released parasitoids at 4 times after the release. When an egg mass was parasitized byT. nubilale, 75.7% of the eggs in the egg mass were parasitized. We developed an equation to estimate the percent of egg masses that a single female was expected to parasitize in a day (efficiency of parasitism) and female disappearance (death and dispersal) rates, if both were constant during our experiment. The exponential disappearance rate was −0.52±0.03 day⁻¹, which implied that 40% of the remaining ΦΦ disappeared per day. The efficiency of parasitism was 0.050% parasitism/Φ/SAI/day, which implied that at least 351,000 ΦΦ/ha would be needed to achieve 90% parasitism. Clearly, forT. nubilale to be a successful biological control agent, efficiency of parasitism must be increased and disappearance rates must be reduced.      

Suppression ofOstrinia nubilalis byTrichogramma nubilale in sweet corn

We conducted inundative release experiments withTrichogramma nubilale (Hymenoptera: Trichogrammatidae) to suppressOstrinia nubilalis (Lepidoptera: Pyralidae) in sweet corn (Zea mays): two experiment duringO. nubilalis first generation and three experiments during second generation. Five measurements of ear and stalk damage were used to assess.O. nubilalis control in treated and untreated plots within each experimental field. In one experiment during second generation, naturalO. nubilalis populations were sufficiently high to demonstrate that the parasitoids (three releases totaling 4.4 million parasitoids per ha) parasitized an estimated 57.4% of the placedO. nubilalis egg masses and reduced the mean number ofO. nubilalis larvae per ear by 97.4% the number of tunnels per stalk by 92.9%, and the number of larvae per stalk by 94.3% in the release plot. Ear damage in this experiment was suppressed to meet acceptable standards for use in cut-corn commercial processing. Larval mortality was apparently density independent, which implies that density-dependent larval loss would not compensate for egg parasitism byT. nubilale.     

Dynamics of Nosema pyrausta in natural populations of the European corn borer, Ostrinia nubilalis: A six-year study

Nosema pyrausta (Paillot) (Microsporida: Nosematidae) is an obligatory intracellular parasite of the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae). This pathogen is maintained in natural populations of O. nubilalis by both horizontal and vertical transmission. The impact of N. pyrausta on fecundity of adults and survival of larvae has been well documented in laboratory and field research. In an extensive study covering a 6 year period at one site, we described the effect of N. pyrausta within O. nubilalis populations in a continuous corn following corn ecosystem. We documented the presence of the pathogen through all life stages of O. nubilalis (egg, larva, pupa, adult), by collecting throughout the crop season and examining each insect stage in the laboratory for the frequency of infection with N. pyrausta. The percentage of infection with N. pyrausta and magnitude of the O. nubilalis population fluctuated throughout generation 1 and generation 2. Both horizontal and vertical transmission played a role in maintaining N. pyrausta in the population in both generations. There were strong correlations between percentage adults with N. pyrausta and percentage larvae with N. pyrausta, and between percentage eggs with N. pyrausta and percentage larvae with N. pyrausta. There was a weak correlation between percentage adults with N. pyrausta and percentage eggs with N. pyrausta. The percentage of insects infected with N. pyrausta was always lowest in the egg.     

Overwintering of the egg parasitoid Trichogramma brassicae in Northern Switzerland

As part of a general risk assessment study weinvestigated the overwintering abilities ofTrichogramma brassicae Bezd. (Hym.: Trichogrammatidae) in Northern Switzerland. Eggs of six host species parasitized by T. brassicae were exposed under outdoor conditions every two weeksbetween 26 September and 7 November 1998. Wefound that T. brassicae is able tooverwinter successfully on six lepidopteranspecies in the families Tortricidae, Noctuidae,Plutellidae, Pyralidae and Crambidae. Between75 and 100% emergence was observed in thefollowing spring for all of the six tested hostspecies exposed on 26 September. On laterexposure dates, spring emergence decreasedsignificantly and no development of T. brassicae offspring occurred from host eggsparasitized on 7 November.Emergence of T. brassicae from eggs ofEphestia kuehniella Zeller parasitized atweekly intervals in autumn 1999 occurred in thesame year until the end of October if the eggswere parasitized by 10 September. All offspringfrom eggs that were parasitized from 17September onwards went into diapause. Springemergence of T. brassicae adults wasobserved between end of April and beginning ofMay both in 1999 and 2000. Fecundity of T.brassicae females that overwintered on E.kuehniella was not significantly different fromthe fecundity of control females that werereared without diapause under optimalconditions at 25 °C. Our resultsdemonstrate that the egg parasitoid T.brassicae is able to overwinter successfullyin Northern Switzerland and has the potentialto establish in Switzerland. As a result,potential non-target effects are not locallyrestricted but may occur on a largegeographical scale.     

Olfactory responses ofTrichogramma maidis Pint,et Voeg.: Effects of chemical cues and behavioral plasticity

Oriented responses of Trichogramma maidisPint, et Voeg. to airborne odors were observed in a four-armed olfactometer. Experiments were carried out with odors of host eggs, the sex pheromone of Ostrinia nubilalisHbn, and maize extract, offered singly or in combination, both to naive wasps and to wasps previously exposed to the tested odor during an oviposition experience. The exploratory behavior in the olfactometer was quantified by means of a computer program which performed a space-time analysis of the insect 's movements. Whereas the naive wasps did not respond to the odor of the eggs, the synthetic sex pheromone of O. nubilalis,or the maize extract presented singly, they did react to a mixture of these three volatile cues. Prior oviposition in the odor of maize extract or in the combination of odors induced an increased preference toward the conditioning scent. This phenomenon did not occur when the wasps were conditioned to egg odor or sex pheromone alone. These results suggest that females can learn to associate some olfactory cues with the presence of the host. Immediately following the presentation of the combination of odors, a strong attraction of experienced wasps occurred; it decreased as the experiment progressed and finally reached the level presented by naive insects. Adult conditioning to the combination of odors also resulted in reduced variability in the behavioral responses.     

Inoculative releases of Trichogramma ostriniae for suppression of Ostrinia nubilalis (European corn borer) in sweet corn: field biology and population dynamics

The effectiveness of inoculative releases of Trichogramma ostriniae Pang and Chen for suppression of Ostrinia nubilalis (Hübner) in sweet corn was assessed. Early-season, low-density (75,000 females ha⁻¹) releases were made, and establishment, levels of parasitism and sex ratios of emerging T. ostriniae quantified. T. ostriniae established effectively for each season that they were released, but appeared to be unable to overwinter. Parasitism levels tracked egg mass numbers closely, and T. ostriniae persisted in fields even where insecticides were applied. Parasitism by indigenous Trichogramma species was 3%. Field populations of T. ostriniae were distinctly female biased (78%), with males produced in the majority of broods. Numbers of males did not increase linearly with number of O. nubilalis eggs parasitized, but appeared to remain constant above an egg mass size of about 20 eggs. A Type-I functional response to increasing egg and egg mass density was found under field conditions, where the proportion of egg masses parasitized remained constant with increasing egg mass density. A relatively consistent percentage of eggs per egg mass was parasitized, with a linear increase in number of eggs parasitized with increasing number of eggs per egg mass. These results show that T. ostriniae established viable reproductive populations in sweet corn following inoculative release, with the potential to contribute to reduced dependence on insecticides for the control of O. nubilalis in an integrated pest management program.     

Changes in host acceptance and host suitability as an effect of rearing Trichogramma maidis on a factitious host

Host acceptance and suitability of Ostrinia nubilalis Hbn. for Trichogramma maidis Pint. et Voeg. (Hymenoptera: Trichogrammatidae) were studied, after rearing the parasitoids on O. nubilalis or Ephestia kuehniella Oliv. (Lepidoptera: Pyralidae) for various numbers of generations. In cages, containing two maize plants with in total 30 egg masses of the natural host (O. nubilalis), wasps continuously reared on either E. kuehniella or O. nubilalis found egg masses in equal proportions. The former however, did not parasitize the eggs successfully, due to either low degree of host acceptance or insufficient host suitability or both. Subsequently, host acceptance behaviour of individual females from different strains was studied by means of direct observations after releasing them into a small arena containing eggs of O. nubilalis or E. kuehniella. Wasps continuously reared on O. nubilalis or 1–3 generations on E. kuehniella accepted egg masses of the natural host better than wasps continuously reared on E. kuehniella. Acceptance of O. nubilalis by the wasps gradually decreased with increasing numbers of generations reared on E. kuehniella. By rearing T. maidis, previously cultured on E. kuehniella, for five generations on O. nubilalis, acceptance of egg masses of this host species did not change. Host suitability of O. nubilalis appears to correspond with host acceptance of the strains. Contrary to the observations on O. nubilalis, acceptance and suitability of E. kuehniella were not influenced by the host on which T. maidis was reared. Results of this study show that host acceptance and suitability of the natural host are important quality factors for T. maidis when mass produced on a factitious host.

---

Zusammenfassung Wirtsakzeptanz und Wirtseignung von Ostrinia nubilalis Hbn. für den Eiparasitoiden, Trichogramma maidis Pint. et. Voeg., wurden untersucht nach der Zucht auf dem natürlichen Wirt, O. nubilalis, oder auf dem Ersatzwirt, der Mehlmotte, Ephestia kühniella Zell.. Versuche mit Käfigen, in denen je zwei Maispflanzen mit insgesamt 30 Eigelegen von O. nubilalis und 40 Weibchen von T. maidis eingesperrt waren, zeigten, dass Weibchen, die seit über 150 Generationen auf O. nubilalis bzw. E. kühniella gezüchtet worden waren, die Eigelege gleich gut fanden. Während die auf O. nubilalis gezüchteten Parasitoide die Eigelege zu 80% parasitierten, war die Parasitierungsrate der auf E. kühniella gezüchteten Parasitoide gleich Null. Die Ursachen dieses Phänomens wurden in weiteren Versuchen durch direkte Beobachtungen über Wirtsakzeptanz und Wirtseignung untersucht. Einzelne Weibchen von Stämmen, die nur auf O. nubilalis oder während einer zunehmenden Generationenzahl auf E. kühniella gezüchtet worden waren, untersuchte man während 15 Minuten unter der Stereoskoplupe in einer kleinen Arena, die ein Eigelege des Maiszünslers oder Eier der Mehlmotte enthielt. Dabei zeigte sich, dass Wespen, die ausschliesslich auf Maiszünslereiern oder nur 1–3 Generationen auf Mehlmotteneiern gezüchtet worden waren, die Eigelege von O. nubilalis besser akzeptierten als solche, die seit vielen Generationen auf E. kühniella vermehrt worden waren. Die Wirtsakzeptanz verschlechterte sich mit zunehmender Generationenzahl auf E. kühniella. Wenn T. maidis nach 23 Generationen auf E. kühniella wieder während 5 Generationen auf O. nubilalis gezüchtet wurde, verbesserte sich die Akzeptanz der Eigelege von O. nubilalis nicht. Die Wirtseignung der Eigelege von O. nubilalis, stimmt überein mit der Wirtsakzeptanz der verschiedenen Stämme. Dies ergaben Versuche mit 15 Minuten Direktbeobachtung und bei 24 Stunden Expositionszeit. Im Gegensatz zu den Beobachtungen auf Eigelegen von O. nubilalis wurde die Akzeptanz und Eignung der Eier von E. kühniella durch die Zucht auf verschiedenen Wirten nicht beeinflusst. Das Ergebnis dieser Untersuchung zeigt, dass Akzeptanz und Eignung des natürlichen Wirtes wichtige Qualitätsfaktoren für T. maidis in der Massenzucht auf Ersatzwirten sind.

     

Inbreeding and incompatibility in Trichogramma nr. brassicae: evidence and implications for quality control

Inbreeding effects and incompatibility relationships were examined in strains of the egg parasitoid Trichogramma nr brassicae (Hymenoptera: Trichogrammatidae) from southeastern Australia. Crosses between strains provided weak evidence of incompatibility in a few cases. However sex ratio in crosses within strains tended to be more female-biased than in crosses between strains. Inbreeding was imposed for four generations (F>0.59) of sib mating. The fitness of inbred strains was compared to that of outbred strains generated by crossing the inbred strains. No effects of inbreeding were found for any of the four female traits examined (fecundity, body length, head width and hind tibia length), indicating that T. nr. brassicae is not subjected to inbreeding depression. Inbreeding effects were also not found for male mating success as expected for the haploid sex. There were differences among strains for all traits apart from fecundity, indicating heritable variation. Strain differences for fitness measures were uncorrelated with wasp size. The potential use of inbreeding in the quality control of Trichogramma for mass-release is discussed. Inbreeding may be a useful tool in minimising the effects of laboratory adaptation, thereby extending the useful life of a strain.     

<Emphasis Type="Italic">Trichogramma Brassicae</Emphasis> and SLAM<Superscript>®</Superscript>, an Integrated Approach to Managing European Corn Borer and Corn Rootworms

European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae); western corn rootworm (WCRW), Diabrotica virgifera virgifera LeConte; and northern corn rootworm (NCRW), Diabrotica barberi (Smith & Lawrence) (Coleoptera: Chrysomelidae) are important pests of corn, Zea mays L., that occur simultaneously in the US Corn Belt. Areawide management strategies for northern and western corn rootworm in Iowa, Kansas, South Dakota, Illinois, and Indiana, and for the Mexican strain in Texas, use SLAM^®, which contains cucurbitacin, a feeding stimulant, and a small amount of the insecticide carbaryl. This management strategy is environmentally sound because it introduces a substantially lesser amount of chemical insecticide into the environment than prophylactic soil insecticide applications for WCRW management. To develop a management program for both corn pests, the compatibility of SLAM^® to control CRW, along with a biological agent to manage European corn borer, was investigated. Laboratory studies were conducted using the egg parasitoid Trichogramma brassicae (Bezdenko) (Hymenoptera: Trichogrammatidae). In experiments in which SLAM^® and T. brassicae were evaluated as components of a management system, SLAM^® did not affect parasitism or parasitoid emergence rates. Trichogramma brassicae displayed vigorous biotic fitness, emergence rates, sex ratios, and fecundity over the duration of the study. Data collected under these laboratory conditions indicated that SLAM^® had no adverse effect on the quality of T. brassicae, indicated by its ability to reproduce in the filial generation.     

Comparative Prey Suitability ofOstrinia nubilalisEggs andAcyrthosiphon pisumforColeomegilla maculata

The effects ofOstrinia nubilalis(Hübner) (Lepidoptera: Pyralidae) eggs andAcyrthosiphon pisum(Harris) (Homoptera: Aphididae), when provided as single prey species and in combination, on life history characteristics ofColeomegilla maculataDeGeer (Coleoptera: Coccinellidae) larvae and adults were quantified. Preimaginal development was not influenced by the larval prey regime; development at 26 ± 1°C was completed in approximately 13.5 days onO. nubilaliseggs,A. pisum,orA. pisumalternated daily withO. nubilaliseggs. The resulting adults weighed 13.0, 10.7, and 12.5 mg when reared onO. nubilaliseggs,A. pisum,andA. pisumalternated daily withO. nubilaliseggs, respectively. Eighteen percent of the individuals died when reared onA. pisum,28% died when reared onO. nubilaliseggs, and 22% died when fedA. pisumalternated daily withO. nubilaliseggs. Seven adult diet combinations, based on diet regimes of larvae and adults, did not cause significant differences in preoviposition period, interoviposition period, and the number of days on which eggs were laid. Total fecundity was influenced both by larval and adult diet. The diet that resulted in highly fecund females wasA. pisumalternated daily withO. nubilaliseggs for larvae andO. nubilaliseggs for adults. FemaleC. maculatafedO. nubilaliseggs had the highest intrinsic rate of increase and net reproductive rate.     

Biology ofTrichogramma ostriniae (Hym.: trichogrammatidae) reared onOstrinia nubilalis (Lep.: pyralidae) and survey for additional hosts

Trichogramma ostriniae has been identified as a candidate for biological control ofOstrinia nubilalis. As little was known about the biology of this parasitoid when reared onO. nubilalis, we undertook experiments to quantify biological parameters important to mass-rearing and ase ofT. ostriniae. When reared continuously onO. nubilalis, femaleT. ostriniae on average lived 2.7 days and produced 24 progeny. Continuous access to honey resulted in a four-fold increase in longevity and fecundity and a significant increase in the percentage of females parasitizing eggs. Rates of fecundity and parasitism decreased with age of female. Likewise, emergence rates and percentage of female progeny decreased with age of parental female.T. ostriniae successfully parasitizedO. nubilalis eggs until the blackhead stage. Most parasitism of eggs and eclosion of adults occurred during the first half of photophase. Eggs of 13 Lepidopterans were parasitized byT. ostriniae. Eggs of the Noctuidae, Pyralidae, and Plutellidae experienced higher levels of parasitism than others tested.T. ostriniae appears to be similar to other species ofTrichogramma in several respects and does not possess any characteristics that limit its potential for mass rearing and use for augmentative biological control ofO. nubilalis.     

Interactions among three trophic levels: the influence of host plant on performance of Pieris brassicae and its parasitoid, Cotesia glomerata

The relative suitability of four host plants was determined both for unparasitized Pieris brassicae L. caterpillars and for Cotesia glomerata (L.) developing in P. brassicae. For unparasitized P. brassicae, growth rate and pupal weight were highest on Brussels sprouts and Swedish turnip, intermediate on rape, and lowest on nasturtium. In contrast, C. glomerata larval developmental rate and adult longevity were greatest for wasps from P. brassicae reared on nasturtium.On all four plants, the host-parasitoid complex attained a lower final weight than unparasitized P. brassicae, and it is argued that this difference is due primarily to reduced consumption by parasitized P. brassicae. Among parasitized caterpillars, however, complex weight was positively correlated with clutch size, suggesting that C. glomerata larvae are able to partially counteract the effect of parasitization per se on host consumption.The host plants of P. brassicae appear to face an evolutionary dilemma: in order to increase the total mass of parasitoids produced, they must suffer greater loss of foliage. This trade-off, if common in nature, may represent a formidable constraint on coevolution between host plants and parasitoids.     

On-farm evaluation of Beauveria bassiana for control of Ostrinia nubilalis in Iowa, USA

Ostrinia nubilalis is (Lepidoptera: Crambidae) a severe pest ofcorn in the major corn growing areas of theUnited States. The efficacy of a Beauveria bassiana application, for season-long suppression of O. nubilalis was evaluated in 1996 and 1997 at locationsacross Iowa. Beauveria bassiana,Mycotech 726 (Mycotech Corporation, Butte, MT)formulated on corn grit granules (14–20 mesh)at 2.2 × 10⁹ conidia/g and applied with ahand-held applicator at the rate of 0.4 g/plant(8.8 × 10⁸ conidia/plant). Applicationswere made when plants were in the V7 or R1growth stage. The length of larval tunneling,percentage of plants not infested with O.nubilalis, percentage of plants with anendophyte, and yield from treated and controlplots were determined. Whorl-stage applicationof B. bassiana in 1996 resulted in asignificant reduction in centimeters oftunneling (46–55%) and the percentageplants not infested by O. nubilalis. In1997, B. bassiana caused significantreductions in larval tunneling at all locations(20–53%); however, a significant increase inthe percentage of plants not infested with O. nubilalis occurred at only one location. Treatment of plants with B. bassiana in1997 did not significantly increase thepercentage of plants with an endophyte;however, the trend, with the exception of onesite, was for a greater percentage ofendophytic plants in treated versus untreatedplots. A whorl-stage application of a granularformulation of B. bassiana was mostefficacious in reducing O. nubilalis larval damage.     

Response of the wasp (<Emphasis Type="Italic">Cotesia glomerata</Emphasis>) to larvae of the large white butterfly (<Emphasis Type="Italic">Pieris brassicae</Emphasis>)

The large white butterfly (Pieris brassicae L) first invaded northernmost Japan from Siberia around 1994, and after a few years, began to expand its range. The wasp, Cotesia glomerata (L) parasitizes larvae of the small white butterfly (Pieris rapae crucivora Boisduval), a usual host in the same geographic area. Some Pieris brassicae larvae in Hokkaido have been parasitized by Cotesia glomerata, but the parasitism rate of Pieris brassicae larvae tends to be lower than that of Pieris rapae. To examine the process of parasitizing Pieris brassicae larvae, we observed how the parasitoid wasp responded to the host larvae on damaged leaves. Cotesia glomerata females tended to avoid Pieris brassicae larvae, and even when female wasps inserted their ovipositors into Pieris brassicae larvae, none laid eggs. The parasitoids obtained from Pieris rapae larvae failed to parasitize Pieris brassicae during the host-acceptance step.     

Odor specificity testing in the assessment of efficacy and non-target risk for Trichogramma ostriniae (Hymenoptera: Trichogrammatidae)

Trichogramma ostriniae Pang and Chen (Hymenoptera: Trichogrammatidae) is an indigenous egg parasitoid of Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) in China. We evaluated T. ostriniae's responses in olfactometer and wind tunnel assays to various host and plant odors that are likely to impact the efficacy and non-target risk of utilizing T. ostriniae as an augmentative biocontrol agent against Ostrinia nubilalis Hübner in the USA. In a Y-olfactometer, female T. ostriniae exhibited innate positive responses to the egg mass volatiles, scale volatiles, and synthetic sex pheromones of O. nubilalis. When exposed to O. nubilalis pheromone while walking on a platform in a wind tunnel, the wasps manifested significant changes in patch exploration behavior, including delayed dispersal from the platform and slowed walking. The wasps did not respond innately to the synthetic pheromone of a non-target species, Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae), however. Exposing wasps to S. frugiperda pheromone together with eggs of a factitious rearing host prior to testing also did not alter the wasps’ lack of response to the pheromone, indicating that associative learning of the novel odor did not occur. Lastly, wasps showed no innate responses to leaf volatiles from corn (Zea mays L.) and pepper (Capsicum annuum L.), two crops attacked by O. nubilalis. We conclude that T. ostriniae is likely to be highly efficient at finding O. nubilalis eggs in the field through the exploitation of host chemical cues. Further, T. ostriniae's response to moth pheromone appears to be relatively host-specific, since the wasps responded to the pheromone of a congener to their natural host, but not to the pheromone of a more distantly related non-target species. This type of odor-specificity could be an important mechanism for reducing the risk of T. ostriniae attack on non-target species.     

Behavioural observations of Pieris brassicae larvae indicate multiple mechanisms of action of analogous drimane antifeedants

We tested 11 analogous synthetic drimane antifeedant compounds for their feeding inhibiting effects on larvae of the large white butterfly Pieris brassicae L. (Lepidoptera: Pieridae) in no-choice tests on the host plant Brassica oleracea L. Furthermore, we observed larval feeding behaviour in no-choice tests to analyze temporal effects of five drimanes. The results show that the five analogous antifeedants differentially influence feeding behaviour and locomotion activity. Warburganal and polygodial are most likely sensory mediated antifeedants. Habituation to these compounds occurs soon after the onset of the tests (i.e., within 0.5–1.5 h). Compound 5 and confertifolin are probably not direct, sensory mediated antifeedants. After 0.5–1.5 h of exposure, these compounds inhibit not only feeding, but also locomotion behaviour, indicating postingestive, toxic effects. Isodrimenin inhibits feeding from the onset of the test and is probably a sensory mediated antifeedant. No habituation occurs to this compound, indicating that isodrimenin is either a very strong antifeedant or that it additionally has postingestive, toxic effects. Topical application of the drimanes on the larval cuticle revealed feeding inhibiting effects, but these could not be related to the occurrence of postingestive feeding inhibiting effects, indicating that this method is inappropriate to show possible postingestive effects of drimanes in P. brassicae. In conclusion, the behavioural observations performed in this research indicate that analogous drimanes inhibit feeding by P. brassicae larvae through multiple mechanisms of action. The results show that, when developing a structure activity relationship (SAR) for a series of antifeedants, it is important to distinguish the mode of action which underlies inhibition of feeding.