In VitroRearing ofTrichogramma minutumRiley (Hymenoptera: Trichogrammatidae) for Ten Generations, with Quality Assessment Comparisons ofin Vitroandin VivoReared Adults

Trichogramma minutumRiley were reared for 10 generations on an artificial diet containing a yeast extract, FreeAmine III, nonfat dry milk, chicken egg yolk, chicken embryo extract, andManduca sexta(L.) egg liquid. Quality control parameters, including adult longevity, sex ratio, pupation rate, percentage of pupae to emerge as adults, adult female body length, number ofHelicoverpa zea(Boddie) eggs parasitized by a female, and percentage of deformed females were assessed and compared to insects rearedin vivoon irradiatedH. zeaeggs. The development time was longer forin vitroreared insects, but there were more deformed females in thein vitroculture. The sex ratio, however, was generally not significantly different between thein vitroandin vivocultures. Thein vitroreared females generally were larger, lived longer, and parasitized moreH. zeaeggs. Emergence of adults was in excess of 75% in all but the firstin vitrogeneration and was generally not significantly different from adult emergence in thein vivoculture. These findings will be of value in the development of a practical system forin vitromass rearing ofTrichogrammafor use in biological control.     

In vitro culture of Trichogramma pretiosum on an artificial medium

The composition of an artificial medium and environmental conditions are described for the in vitro rearing of the egg parasite Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). The medium was composed of defined amounts of protein, carbohydrates, lipid, salts, and vitamins, but also contained up to 40% insect hemolymph. The hemolymph was necessary to induce pupation. T. pretiosum eggs were obtained by dissection of Heliothis virescens (F.) (Lepidoptera: Noctuidae) eggs. In vitro reared T. pretiosum were similar in size to H. virescens reared T. pretiosum, and females were fecund.

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Résumé Les oeufs de Trichogramma pretiosum ont été obtenus par dissection d'oeufs d'Heliothis virescens. T. pretiosum Riley (Hymenoptère, Trichogrammatidae) a été élevé avec succès sur un substrat synthétique. Outre des quantités définies de protéines, glucides, lipides, éléments minéraux et vitamines, la ration contenait aussi jusqu'à 40% d'hémolymphe de Manduca sexta. L'hémolymphe était nécessaire pour induire la nymphose. En plus de la nourriture, les conditions d'environnement sont apparues extrêmement importantes pour élever T. pretiosum dans des conditions satisfaisantes. Le contrôle de l'humidité relative, en particulier, était le facteur le plus important. Les adultes produits au cours de cette étude étaient d'apparence normale; ils se sont accouplés sans problèmes, les femelles étaient fécondes et leur taille ne différait pas de celle d'individus élevés sur H. virescens.

     

Isoenzyme patterns and biology ofTrichogramma minutum as influenced by rearing temperature and host

Females ofTrichogramma minutum Riley reared at temperatures below 17°C had lower percent emergence, lower fecundity on the first day of emergence, and a reduced rate of population increase (r_i) than those reared at temperatures above 17°C. Parasitoids were more fecund and lived longer when raised onChoristoneura fumiferana (Clemens) compared toEphestia kuehniella (Zeller). The parasitoids performed better when parasitizing the same species of host on which they had been reared. Isoenzyme patterns were not affected whenT. minutum was reared at 13 or 27°C but additional bands appeared when parasitoids were reared onSitotroga cerealella (Olivier) compared toC. fumiferana. No correlation could be made between biological and biochemical changes inT. minutum. Isoenzymes have the potential to assess the quality of parasitoids during mass production.

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Einfluss von Zuchttemperatur und Wirt auf Isoenzymmuster und Biologie vonTrichogramma minutum Riley

Zusammenfassung Weibchen vonTrichogramma minutum, welche bei 17°C gezüchtet wurden, hatten eine geringere Schlüpfrate, eine geringere Fruchtbarkeit am ersten Tag nach dem Schlüpfen sowie ein reduziertes Populationswachstum (r_i) im Vergleich zu Tieren welche sich bei Temperaturen über 17°C entwickelt hatten. Wurden die Parasitoiden aufChoristoneura fumiferana gezogen, so war ihre Fruchtbarkeitsrate wie auch Lebensdauer grösser als wennEphestia kuehniella als Wirt diente. Die parasitische Fähigkeit vonT. minutum war immer auf derjenigen Wirtsart am grössten auf der die Schlupfwespen vorher gezogen wurden. Aufzuchttemperaturen von 13°C oder 27°C hatten keinen Einfluss auf den Isoenzymbändern wenn die Tiere statt aufC. fumiferana aufSitotroga cerealella gezogen wurden. Es konnte aber kein Zusammenhang zwischen den biologischen und biochemischen Veränderungen gefunden werden. Hingegen besteht die Möglichkeit während der Massenproduktion die Qualität der Parasitoiden anhand von Isoenzymprofilen zu überwachen.

     

Refinement of a cell line based artificial diet for rearing the parasitoid wasp, Trichogramma pretiosum

An artificial diet incorporating insect cells originally developed for Trichogramma australicum Girault (Hymenoptera: Trichogrammatidae) was successfully used to rear Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). To refine the diet, individual components were removed. Chicken egg yolk and the insect cells were identified as the most important components for T. pretiosum development. Their removal resulted in few pupae and no adults. Removal of Grace’s insect medium, a common component of artificial diets, was found to markedly improve the development of T. pretiosum, producing 60% larva to pupa transition and 19% pupa to adult transition. There was no significant difference in T. pretiosum development on diets in which milk powder, malt powder or infant formula were interchanged, despite differences in nutrient composition. The use of yeast extract resulted in significantly higher survival to the adult stage when compared with yeast hydrolysate enzymatic and a combination of yeast extract and yeast hydrolysate enzymatic. Comparison of four antimicrobial agents showed the antibacterial agent Gentamycin and the antifungal agent Nystatin had the least detrimental effect on T. pretiosum development. The use of insect cell line diets has the potential to simplify artificial diet production and significantly reduce T. pretiosum production costs in Australia compared to diets using insect hemolymph or the use of natural or factitious hosts.     

Oviposition and reproductive performance of a generalist parasitoid (Trichogramma pretiosum) exposed to host species that differ in their physical characteristics

The response of generalist egg parasitoids to alternative natural hosts that are present simultaneously is not well known. We investigated the behavior of Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) in relation to two field hosts Helicoverpa armigera Hübner and Spodoptera litura Fabricius, in choice and no choice tests. We quantified the effects of natal host species and post-emergence adult age on the oviposition preference of the parasitoids. H. armigera eggs were consistently preferred over S. litura eggs, regardless of the natal host and adult age. When only S. litura eggs were available as hosts, they were parasitized at statistically similar rates to H. armigera eggs (average of 17 ± 2.7 vs. 13 ± 3.0, H. armigera to S. litura). The adult lifespan and lifetime fecundity of T. pretiosum were variable but were affected by natal host species and/or host species to which they were exposed. Mean lifespan and fecundity of parasitoids that had developed in H. armigera eggs and were exposed to H. armigera eggs for oviposition were 13.9 ± 1.8 days and 98.7 ± 11.0 adult offspring. By contrast, those that developed in S. litura eggs and were exposed to S. litura eggs for oviposition lived for 7 ± 0.9 days and produced 53.8 ± 8.0 adult offspring. The ovigeny index (OI) was significantly lower in the parasitoids exposed to H. armigera eggs than in those exposed to S. litura eggs, regardless of the natal host, indicating that H. armigera eggs sustain the adult parasitoids better than S. litura eggs. These results are used to predict parasitoid behavior in the field when both hosts are available.     

Response of the egg parasitoid Trichogramma pretiosum to the sex pheromone of its host Heliothis zea

This paper presents results of olfactometer experiments with the egg parasitoid Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) and its host the corn earworm moth, Heliothis zea (Boddie) (Lepidoptera: Noctuidae). The sex pheromone of the host significantly reduced the total number of border crossings between odour fields in the olfactometer. Also, female parasitoids made significantly more visits to the calling moth odour field than to the opposite control field in the olfactometer. Further, the wasps spent significantly more time in the olfactometer field containing the sex pheromone released by calling virgin moths, than in control fields. If non-calling virgin moths were used as odour source, the response was reversed and wasps were repelled by the odour of the moths, and the numbers of visits were evenly distributed over the four flow fields. These results are discussed in the context of foraging ecology of egg parasitoids.

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Résumé Des expériences menées en olfactométrie avec le parasite oophage Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) et son hôte, Heliothis zea (Boddie) (Lepidoptera: Noctuidae) donnent les résultats suivants. La présence de la phéromone sexuelle de l'hôte réduit de façon significative le nombre de passages entre les champs odorisés. De mème, les parasites visitent plus fréquemment le champ qui dispense l'odeur de la femelle en appel que le champ témoin placé à l'opposé. Par ailleurs, le temps passé dans le champ qui contient la phéromone sexuelle émise par les femelles vierges en appel est significativement supérieur à la durée de visite des champs témoins. Si l'on utilise comme source d'odeur des femelles vierges qui ne sont pas en appel, la réponse est inversée, les parasites sont alors repoussés par l'odeur de ces papillons et le nombre de visites est distribué de façon aléatoire entre les quatre champs. Ces résultats sont discutés dans le contexte de l'écologie du comportement de recherche chez les parasites oophages.

     

Comparative Development and Performance of Artificially Reared versus Host-RearedDiapetimorpha introita(Cresson) (Hymenoptera: Ichneumonidae) Wasps

Diapetimorpha introita(Cresson) (Hymenoptera: Ichneumonidae), a native ectoparasitoid ofSpodopteraspp. pupae, was reared in the laboratory on an artificial diet devoid of any insect host components. Diet-reared wasps demonstrated a propensity to search for and parasitize natural hosts in a field cage trial. Longevity of the diet-reared wasps was comparable with the longevity of wasps reared on host pupae. Survival rate ofD. introitawas 61.3% when reared on diet and 76.3% when reared on host pupae. Wasps reared on the artificial diet had longer developmental times, reduced fecundity, and reduced adult weights compared to wasps reared on host pupae. These studies suggest that future research efforts should focus on increasing fecundity and weight of diet-reared wasps and decreasing the mortality and developmental time of wasps reared on the artificial diet. The ability to rearD. introitaon an inexpensive, artificial diet significantly enhances the potential of mass rearing this parasitoid for inundative releases againstSpodopteraspp.     

Anin vitro rearing system for the propagation of the ectoparasitoidCatolaccus grandis

An experimental ‘closed’ rearing system, where egg and larval manipulations were eliminated, was developed for the in vitro rearing of Catolaccus grandis (Hymenoptera: Pteromalidae) Burks, an important ectoparasitoid of the cotton boll weevil, Anthonomus grandis Boheman. In this rearing model, n-hexane (a synthetic ovipositional stimulant for this parasitoid), was smeared on the Parafilm® cover of a modified rearing chamber (a Multiwell®) tissue culture plate) to induce the deposition of uncontaminated eggs, on the inner side of this waxy membrane, and on or around an agar retained diet that had been dispensed into the individual chamber wells. When the efficiency of the in vitro rearing system was compared to the current in vivo rearing method for this species, the duration of the life cycle was significantly shorter in parasitoids reared in vivo, but this difference was less than one day (17.8 vs 17.1 days, respectively). On the other hand, the number of eggs laid in the in vitro rearing chamber during a 4 h period was c. 2.5 times greater than in the conventional in vivo rearing apparatus, and adult yields were c. 25% greater when using the in vitro closed rearing method. Male to female ratios were c. 1:9 when reared in vitro as compared to 1.0:1.5 for those reared in vivo. There were no apparent adverse effects of this in vitro rearing system on the parasitoid's general behavior and reproduction after two consecutive generations.     

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.

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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.

     

Effects of fine-grain habitat complexity on egg parasitism by three species of Trichogramma

Effects of microhabitat complexity on host foraging by three species of Trichogramma (T. deion, T. ostriniae, and T. pretiosum) were evaluated under simulated stored product conditions. All three species have been considered as potential biological control agents for the Indianmeal moth, Plodia interpunctella, in retail stores and warehouses. Trials were conducted with single naïve female parasitoids in 10-cm Petri dishes that were either empty, contained flour, or contained millet. Empty 15-cm Petri dishes, which served as a surface area control, comprised a fourth treatment. Females were allowed to forage for sentinel egg disks for 2 h, after which percentages of parasitism and egg mortality were computed. In addition, behavioral observations were made on a subset of the trials. Trichogramma deion parasitized more eggs than the other two species in the empty dishes and in the dish containing flour. For T. deion, rates of parasitism and egg mortality were significantly greater in both the small and large empty dishes than in the small dishes containing flour or millet. Parasitism was consistently low for all three species in the grain-filled dishes. Among species, T. ostriniae spent the most time walking, while T. pretiosum was the most sedentary. In addition, T. pretiosum spent significantly more time on the first egg visited compared with subsequent eggs. T. deion may be the best-suited for use as a biological control agent for P. interpunctella. However, the potential negative effects of fine-grain habitat complexity should be taken into account when developing a release protocol for Trichogramma spp.     

Variation in locomotion between laboratory strains of Trichogramma maidis and its impact on parasitism of eggs of Ostrinia nubilalis in the field

The locomotion of females of different strains of the parasitic wasp Trichogramma maidis (Hymenoptera, Trichogrammatidae) was measured under standardized conditions and compared to their efficiency in the field for several years. A manual and an automatic method for recording and analysing the locomotion of insects are described and results for differently reared laboratory strains of T. maidis are presented. The correlation between manual and automatic measurements of the travel speed was significant (r=0.999). The automatic method was developed by means of an Image Processing System. T. maidis females were individually released in an arena, and the walking tracks were recorded by a video camera. To reconstruct the real path, a mathematical model which gives highly accurate path length estimations — termed arcus interpolation — was used. Furthermore the same programme permits the calculation of parameters like turning rate and angles, the activity pattern over the whole observation period or for selected parts of it. The locomotion of different T. maidis strains in the laboratory was compared with their efficiency in the field. An obvious relation between locomotion and parasitization in the field was apparent. The probability for parasitism of Ostrinia nubilalis Hbn. (Lepidoptera, Pyralidae) egg batches decreased linearly with increasing distance from the release point of Trichogramma wasps. The travel speed of the females of different T. maidis strains was found to be positively related to the potential for parasitism in the field. The travel speed can therefore be used as a parameter to estimate the capacity for host location and the efficiency of a T. maidis strain for inundative biological control programmes.

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Zusammenfassung Während mehrerer Jahre wurde die im Labor bestimmte Laufgeschwindigkeit von Trichogramma maidis Pint. & Voeg. (Hymenoptera, Trichogrammatidae) mit ihrem Parasitierungspotential im Freiland verglichen. Für die Bestimmung der Bewegungsaktivität von T. maidis werden zwei Methoden beschrieben. Mit der einen wird die zurückgelegte Wegstrecke eines Insekts, während einer definierten Zeitperiode, mit einem Planimeter gemessen. Für die zweite, automatische Bestimmung der Bewegungsaktivität verwendete man einen IPS (Image Processing System von KONTRON München), für welches entsprechende Programme entwickelt wurden. Bei der zweiten Messmethode wird der Weg jedes einzelnen Tieres in einer Laufarena von 24 mm Durchmesser während 10 Min mit einer Videokamera aufgenommen. Jede Sekunde werden die Schwerpunktskoordinaten des Tieres bestimmt. Die Länge des zurückgelegten Weges der Tiere wurde mit Hilfe eines Berechnungsmodells geschätzt. Die Wegstreckenkorrektur erfolgte mit Kreisbögen, welche aus drei aufeinanderfolgenden Punkten jedes Wegabschnittes berechnet werden. Mit dieser Korrektur konnte auf einfache Weise eine sehr genaue Schätzung der Laufstrecke erzielt werden. Die Ergebnisse der beiden Methoden weisen eine enge Korrelation (r=0.999) auf. Mit dem entwickelten Programm lassen sich zusätzlich, zu den effektiven Laufstrecken, auch noch Anzahl und Stärke der Richtungsänderungen sowie das Aktivitätsmuster der ganzen Beobachtungsperiode bestimmen.Die Untersuchungen ergaben, dass die Parasitierungsrate der Eigelege von Ostrinia nubilalis Hb. (Pyralidae, Lepidoptera) mit zunehmender Distanz vom Freilassungspunkt von T. maidis abnahm. Die Wahrscheinlichkeit für O. nubilalis-Gelege parasitiert zu werden, kann innerhalb der ersten 8 m am besten mit einem linearen Modell beschrieben werden. Das Parasitierungspotential definierte man als Integral der Parasitierungs-Wahrscheinlichkeit für O. nubilalis-Gelege über die Distanz vom Freilassungspunkt weg. Es zeigte sich, dass zwischen den im Labor bestimmten Laufgeschwindigkeiten der verschiedenen T. maidis Stämme und den im Feld gemessenen Parasitierungspotentiale eine positive Beziehung besteht. Die Laufgeschwindigkeit von T. maidis Weibchen kann somit als ein Parameter zur Bestimmung der Suchkapazität eines Trichogramma Stammes und in etablierten Bekämpfungsprogrammen als Eignungstest der Provenienzen für die biologische Bekämpfung betrachtet werden. Laufgeschwindigkeits-Bestimmungen ermöglichen es, rasche und ökonomische Qualitätskontrollen von in Massen gezüchteten T. maidis vorzunehmen, indem bereits im Labor die Eignung der T. maidis Stämmen für die Bekämpfung von O. nubilalis beurteilt werden kann.

     

Preliminary results on in vitro rearing of the endoparasitoid Cardiochiles nigriceps from egg to second instar

The composition of an artificial medium and technical procedures used for in vitro rearing of the endophagous larval parasitoid Cardiochiles nigriceps Viereck (Hymenoptera, Braconidae), from post-germ band egg to the 2nd instar larva, are described. Amino acids, carbohydrates, salts, and vitamins were supplied in defined amounts as an aqueous solution which, when supplemented with 20 mg/ml of bovine albumin, 5 mg/ml of lactalbumin (enzymatic hydrolysate), 20% (v/v) fetal bovine serum, 20% (v/v) milk and 10% (v/v) chicken egg yolk, allowed for parasitoid growth and molting to the 2nd instar. Molting to the final instar was never observed.     

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.     

Partial In Vitro Digestion of Active Gliadin-Related Peptides in Celiac Disease

Peptides corresponding to residues 75–86 (RPQQPYPQPQPQ) and 75–85 of the A-gliadin structure, which were shown to be active in an animal model of celiac disease, were digested in vitro with small intestinal mucosa from children with celiac disease in remission and with mucosa from normal children. The products of digestion were separated into two fractions by gel permeation chromatography. Undigested residues (M_r > 400 fraction) from both peptides contained mainly glutamine, proline, and tyrosine, while the digested materials (M_r < 400 fraction) contained mainly proline, glutamine and arginine. Much larger amounts of undigested peptides were obtained from digestion with celiac mucosa than from normal mucosa. The results with peptide 75–86 indicated that the octapeptide 77–84 (QQPYPQPQ) was the main residual component and this peptide was shown to be active in the assay. Peptide 77–84 was also obtained as a residue from digestion of peptide 75–85, together with heptapeptide 77–83. The results lend further support for a primary mucosal defect in celiac disease and indicate that residual peptides in the small intestine of patients with the disease still retain appreciable toxicity.     

Development of an artificial host egg for in vitro egg laying of Trichogramma galloi and T. pretiosum using plastic membranes

An artificial host egg was developed as a first attempt to improve the techniques for in vitro rearing of Trichogramma galloi Zucchi and T. pretiosum Riley (Hymenoptera, Trichogrammatidae). Plastic membranes (polyethylene, polypropylene and polystyrene) of different thickness were tested for their usefulness in manufacturing artificial eggs, using a thermal system to produce semi-spherical cupules on the plastic surface. These artificial eggs were filled with larval hemolymph of Helicoverpa zea (Boddie) (Lepidoptera, Noctuidae), and the optimal size of the artificial egg and density of females/artificial egg for the highest host acceptance (percentage of artificial eggs with at least one parasitoid egg) as well as the ideal level of parasitism (eggs/artificial egg) were determined. High-density polyethylene 7–8 and 9–10 m thick resulted in the best acceptance and level of parasitization for both parasitoid species. Six females per artificial egg resulted in the best rate of parasitization for these parasitoids, although these data were not different when using 4 or 5 females of T. galloi. The size, shape and surface texture of the artificial eggs were found to provide enough cues to elicit parasitization behavior in Trichogramma females, since no chemical stimulus was provided.     

The effect of host size on quality attributes of the egg parasitoid,Trichogramma pretiosum

In a study of the quality ofTrichogramma pretiosum Riley (Hymenoptera: Trichogrammidae), we compared female wasps emerging from natural hosts, parasitized in the laboratory or the field with those emerging from factitious hosts used for commercial mass production. Females from the natural hosts were larger, more fecund, and longer lived than those from the factitious hosts. Compared to small females, large female wasps are substantially more fecund when honey (carbohydrate) is available but marginally more fecund when honey is unavailable. The size of a femaleT. pretiosum depends on two factors: the size of the host egg from which it emerges even when the wasp was gregarious, and the number of conspecifics that emerge with it. The similarities in the size distribution of female wasps emerging from natural hosts, in conjunction with the mechanism by whichTrichogramma measure host size and allocate eggs accordingly, suggests the hypothesis that size related components of fitness in femaleT. pretiosum are under strong selection in the field.     

In vitro rearing ofExorista larvarum on tissue culture-based diets

Exorista (=Tachina) larvarum (L.) (Diptera, Tachinidae), a polyphagous parasitoid that attacksLymantria dispar L. andHyphantria cunea (Drury), was rearedin vitro from egg to adult on four tissue culture media-based diets (TMM-FH, SCHNEIDER'S, EX-CELL 400, and SF-900). The kind of tissue culture media in the diets did not influence the adult yield (34 to 55%) and puparium weight (26–27 mg). Adult yield and the puparium weight ofE. larvarum developed on TNM-FH and SCHNEIDER'S-based diets containing different amounts ofGalleria mellonella pupal extract (PE) (0, 1.25, 2.5 and 5%), were lower on diets without PE. In diets without PE development times from oviposition to adult emergence, were shorter on TNM-FH (19 days) than on SCHNEIDER'S-based diet (25–26 days). The adults that developed on artificial diets were able to parasitize the factitious hostG. mellonella and produce viable progeny. The results demonstrate thatE. larvarum is the most promising parasitoid ever studied forin vitro mass production.