Piero Baronio

Dependence of the parasitoid Gonia cinerascens on the hormones of its lepidopterous hosts

Abstract Development of first instar larvae of Gonia cinerascens, which rest in the muscles of host caterpillars, is triggered by the release of the hosts ecdysteroids when the juvenile hormone is absent. Ecdysteroids act on the parasitoid directly and at the same time induce physiological and biochemical changes in the host, which are indispensable for the parasitoids development. These changes do not occur when metamorphosis of the host is suppressed with the juvenile hormone. Normally the parasitoids initiate development at the larval-pupal transformation of the host, but under experimental conditions, they do so whenever a high ecdysteroid titre is coupled with the proper internal environment in the host, that is in decapitated caterpillars, isolated host abdomens, and when implanted into host pupae. Activated parasitoids moult into the second instar and migrate to the exuvial space of the host; this migratory behaviour is also triggered by ecdysteroids and may be induced experimentally in the first instar parasitoids. Unknown clues direct the migrating parasitoids under the wings and appendages of the host pharate pupal stage. The second instar parasitoids, which anchor to the integument of the host pupae, apparently develop independently of the hosts hormones: they can produce third instar larvae, pupae, and adult flies when cultured in vitro.

Biological and biochemical characteristics for quality control of Harmonia axyridis (Pallas) (Coleoptera, Coccinellidae) reared on a liver-based diet

Biological and biochemical parameters of a flightless strain of Harmonia axyridis, fed on a pork liver-based artificial diet and on Ephestia kuehniella eggs as controls, were compared. The diet-grown larvae showed a significantly longer developmental time and a lower adult emergence rate compared to control larvae. The weights of the newly emerged adults were significantly higher for adults fed E. kuehniella eggs during their larval stages than fed the artificial diet. In contrast, larval food source had no effect on the duration of the pre-oviposition period or adult longevity. For adults fed on E. kuehniella eggs as larvae, a significantly longer pre-oviposition period, lower daily weight gain and fecundity were found for the diet-fed females compared to those fed on E. kuehniella eggs throughout the life span. The adult food source had no significant effect on longevity and fertility. Lower amino acid and fatty acid contents (in particular C16:1 and C18:3n-3) were found for the prepupae and newly emerged females obtained from diet-reared larvae compared to controls. Deficiencies in fatty acids C16:1 and C18:3n-3 were also observed in females obtained from E. kuehniella egg-reared larvae and fed on diet from adult emergence. The analyses of the foods showed deficiencies in artificial diet, especially for some amino and fatty acids. The results suggest a non-optimal composition of the artificial diet and some possibilities for its improvement. However, this polyphagous predator could be reared from first instar larvae to fully reproductive adults on a pork liver-based artificial diet.

Effects of conventional and transgenic Bacillus thuringiensis galleriae toxin on Exorista larvarum (Diptera: Tachinidae), a parasitoid of forest defoliating Lepidoptera

Abstract The Cry9Aa entomocidal toxin from Bacillus thuringiensis ssp. galleriae (Btg) and an epiphytic Pseudomonas sp. derivative carrying the cloned cry9Aa gene from Btg are active against the pine processionary moth Thaumetopoea pityocampa and the laboratory model species Galleria mellonella. A laboratory study was conducted to investigate the side effects of the Cry9Aa toxin and the engineered bacterium on the post-embryonic development of Exorista larvarum, a larval parasitoid of forest lepidopterous defoliators, cultured in the factitious host G. mellonella. In a first experiment, the purified toxin and the commercial Bt preparation Foray 48B induced a mortality of G. mellonella sixth-instar larvae significantly higher than that of the distilled water control. In parallel, the development of E. larvarum in this host was assessed, but no significant difference was found for any of the parasitoid parameters examined (i.e., eggs oviposited, percentage of puparia and adults and puparial weights). In subsequent experiments, cry9Aa-Pseudomonas suspension significantly increased the mortality of sixth instar G. mellonella larvae compared to untransformed Pseudomonas sp. suspension and distilled water. As to the parasitoid parameters, the cry9Aa-Pseudomonas did not significantly affect the number of oviposited eggs, percentage of puparia and puparial weights. It can be concluded that the post-embryonic development of E. larvarum was not affected by host treatment with either Cry9Aa toxin or cry9Aa-Pseudomonas under the laboratory conditions tested. Although direct effects on parasitoid performance have not been shown, indirect effects could still occur and need to be considered in future studies concerning the effects of genetically modified Bt-derivatives.

In vitro rearing of the parasitoid Exorista larvarum (Diptera: Tachinidae) from eggs laid out of host.

We evaluated the possibility of using the eggs laid out of host to rear Exorista larvarum (L.) (Diptera: Tachinidae), a larval parasitoid of Lepidoptera, on artificial media. In a first experiment, eggs oviposited on a plastic sheet (either by inexperienced or experienced females) showed the same in vitro hatching capability as those removed from the larvae of the factitious host Galleria mellonella L. Subsequently, eggs laid on the host integument or out of host, either by inexperienced or experienced females, were removed from the oviposition substrate and placed on a skimmed milkbased artificial medium. The percentages of hatched eggs, of puparia and adults, as well as the puparial weights did not differ significantly among the three treatments. These findings suggested that E. larvarum may be successfully reared in vitro with a total exclusion of the host insect. In a further test, no difference for the in vitro hatching time was found between the eggs oviposited either on a plastic sheet or on G. mellonella larvae in the same length of time (60 min). This result suggested that at oviposition the out-of-host eggs were unincubated, similarly to those that had been laid on the host larvae.

The Influence of Pine Volatile Compounds on the Olfactory Response by Neodiprion sertifer (Geoffroy) Females

Females of the pine sawfly Neodiprion sertifer (Hymenoptera Diprionidae) usually avoid Pinus pinea trees as host plants. In contrast, this sawfly species is highly attracted by P. sylvestris and P. nigra trees. Here, we investigated which pine volatiles might mediate this behavior by in situ sampling experiments and olfactometer laboratory tests. Volatiles emitted from P. pinea, P. sylvestris, and P. nigra foliage were sampled by solid-phase microextraction (SPME). Analysis of these volatiles by coupled gaschromatography/mass spectrometry revealed that the relative amounts of the compounds emitted by the three species were significantly different. A discriminant analysis showed that the amounts of limonene and myrcene significantly contributed to the species-specific volatile patterns. Pinus pinea emitted higher relative amounts of limonene than the other pine species. Pinus sylvestris emitted the highest relative amounts of myrcene. When testing the response of N. sertifer females to these pine terpenoids in an olfactometer bioassay, a low amount of limonene was attractive, while a repellent effect was evident when higher amounts were used. The sawfly females showed no significant olfactory response to myrcene. These data suggest that low relative amounts of limonene have a significant function in attracting N. sertifer females, while high amounts might contribute to avoidance of a tree.

A technique for continuous rearing of Neodiprion sertifer (Geoffroy) (Hym., Diprionidae) in the laboratory

The laboratory rearing of Neodiprion sertifer (Geoffroy) on Pinus sylvestris L. plants grown in vases was successfully achieved on a continuous basis. Larvae were seen to emerge on average after 106 days from egg clusters being kept on plants held at a temperature of 15 ± 1°C and with a 10‐h photophase until emergence. These larvae were then transferred to plastic containers and kept at a temperature of 20 ± 1°C with a 12‐h photophase where, under optimum hygiene conditions and being fed fresh pine branches, they completed their development on average in 30.6 days. Adults began emerging from the cocoons after about 30 days, continuing to do so for at least 7 months. Under these conditions the insect completes its life cycle in 179 days, thus permitting to obtain two generations a year in the laboratory. This staggered adult emergence pattern makes specimens available at all stages of development for experimental purposes throughout most of the year.