Codeine accumulation and elimination in larvae, pupae, and imago of the blowfly Lucilia sericata and effects on its development.

The aim of this study was to evaluate the reliability of insect larvae as samples for toxicological investigations. For this purpose, larvae of Lucilia sericata were reared on samples of minced pig liver treated with different concentrations of codeine: therapeutic, toxic, and potentially lethal doses. Codeine was detected in all tested larvae, confirming the reliability of these specimens for qualitative toxicology analysis. Furthermore, concentrations measured in larvae were correlated with levels in liver tissue. These observations bring new elements regarding the potential use of opiates concentrations in larvae for estimation of drug levels in human tissues. Morphine and norcodeine, two codeine metabolites, have been also detected at different concentrations depending on the concentration of codeine in pig liver and depending on the substance itself. The effects of codeine on the development of L. sericata were also investigated. Results showed that a 29-h interval bias on the evaluation of the larval stage duration calculated from the larvae weight has to be considered if codeine was present in the larvae substrate. Similarly, a 21-h interval bias on the total duration of development, from egg to imago, has to be considered if codeine was present in the larvae substrate.

Susceptibility of different insect pupae to the bacterial symbiont, Xenorhabdus nematophila, isolated from the entomopathogenic nematode Steinernema carpocapsae

Cells of the bacterial symbiont Xenorhabdus nematophila from the entomopathogenic nematode, Steinernema carpocapsae entered the pupae of Plutella xylostella after 15 minutes treatment with suspensions containing the bacterial cells. Secretions of Xenorhabdus nematophila, in either broth or water, were found lethal to the pupae of P. xylostella when applied in moist sand. The bacterial symbiont Xenorhabdus nematophila was found lethal to the pupae of greater wax moth (Galleria mellonella), beet armyworm (Spodoptera exigua), diamondback moth (Plutella xylostella) and black vine weevil (Otiorhynchus sulcatus) in the absence of the nematode vector and the cells of X. nematophila entered the haemocoele of the pupae.

Use of entomopathogenic bacterium Pseudomonas putida (Enterobacteriaceae) and it secretions against the Greater Wax Moth Galleria mellonella pupae

The bacterium from Pseudomonas putida from Steinernema abbasi and its metabolic secretions caused the mortality of the Galleria mellonella pupae. Experiments were conducted in sand and filter paper on time exposure, temperature, moisture, dose and time of penetration of bacterium in pupae and tested stored or dried toxic metabolites using G. mellonella pupae as a test target organism. Death of pupae was probably due to the toxic metabolites. Pseudomonas putida cells were recovered from the haemocoele when bacterial cells were applied to the G. mellonella pupae indicating that bacterial cells can enter the haemocoele in the absence of nematode vector. Penetration of bacterium was found rapidly after application on G. mellonella pupae. Pseudomonas putida or its toxic secretions can be used as a microbial control for insect control. The experimental results indicate that there is possibility of using P. putida and its toxic secretions as a biopesticide and can contribute in the development of new microbial and biological control against insect pests.

Influence of temperature on the production and infectivity of entomopathogenic nematodes against larvae and pupae of vine weevil, Otiorhynchus sulcatus, (Coleoptera: Curculionidae)

Susceptibility of late instar vine weevil Otiorhynchus sulcatus larvae and pupae to four species entomopathogenic nematodes were tested. Bioassays on production and infectivity to larvae and pupae were compared for two steinernematids and two heterorhabditis such as Steinernema carpocapsae, S. feltiae, Heterorhabditis indica and H. bacteriophora. Nematodes production of all species was determined by the number infective juveniles (IJs) established in vine weevil larvae and pupae O. sulcatus using sand and filter paper bioassay. S. feltiae produced the maximum number in larvae and pupae at 20°C as compared to other nematodes but production of H. indica, was better at 25°C in larvae and pupae followed by H. bacteriophora, S. carpocapsae and Infectivity test of larvae and pupae was also done in sand media. Infective juveniles recovered from larvae and pupae when infected with S. feltiae produced maximum infective juveniles at 20°C temperatures than all other isolates. H. bacteriophora produced higher number of IJs in larvae and pupae than all other nematode isolates at 25°C. This paper indicates the application of nematodes with the knowledge of insect pest biology represents a possible new strategy for O. sulcatus larvae and pupae.

Entomopathogenic fungal activity against pupae and adult Haematobia irritans (Diptera: Muscidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Palmistichus elaeisis (Hymenoptera: Eulophidae) parasitizing pupae of Citioica anthonilis (Lepidoptera: Saturniidae) collected on Piptadenia gonoacantha (Fabaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of the juvenile hormone on the development of the mandibular gland in workers' pupae of Apis mellifera L. (Hymenoptera, Apidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mispatterning in the ommatidia of Apis mellifera pupae treated with a juvenile hormone analogue

To further understand the function of morphogenetic hormones in honeybee eye differentiation, the alterations in ommatidial patterning induced by pyriproxyfen, a juvenile hormone (JH) analogue, were studied by scanning and transmission electron microscopy. Prepupae of prospective honeybee workers were treated with pyriproxyfen and the effects on ommatidial differentiation were described at the end of the pupal development. The results show that the entire ommatidia, i.e., the dioptric as well as the receptor systems, were affected by the JH analogue. The wave of ommatidial differentiation, which progresses from the posterior to the anterior region of the pupal eyes, was arrested. In treated pupae, the rhabdomeres only differentiated at the apical axis of the retinula, the secondary and tertiary pigment cells did not develop their cytoplasm protrusions, and the cone cell quartet did not pattern correctly. Simultaneously, an intense vacuolization was observed in cells forming ommatidia. In a previous study we showed that pyriproxyfen exerts an inhibition on pupal ecdysteroid secretion. In this sense, the arrested ommatidial differentiation in pyriproxyfen-treated pupae could be due to a secondary effect resulting from an alteration in pupal ecdysteroid titers. J. Morphol. 249:89-99, 2001. (C) 2001 Wiley-Liss, Inc.

Induction of the split sting trait in Africanized Apis mellifera (Hymenoptera : Apidae) by cold treatment of pupae

Split sting is the name given to a nonfunctional honey bee sting characterized by lancets not attached to the stylet. It has appeared in a mutant line in Brazil, and has provoked interest as a possible means to reduce honey bee colony defensiveness. We induced this alteration in Africanized Apis mellifera L. workers and queens by maintaining pupae at 20 degrees C. In particular, we determined the pupal phase most susceptible to alterations in the sting caused by cold treatment, and we investigated whether this treatment also affected survival to the adult phase and wing morphology. The highest frequency of split sting was detected in workers treated at the pink-eyed pupal phase. The lowest frequency was observed in the bees treated at the oldest worker pupal phase studied (brown-eyed pupae with lightly pigmented cuticle). Both queen pupal phases tested (white and pink-eyed pupae) were equally sensitive and produced high percentages of adults with split sting. However, the 20 degrees C treatment of workers and queens, at the different pupal phases, resulted in high frequencies of adults with deformed wings. Also, fewer workers and queens treated at the earlier pupal stages reached adult emergence. There was also an arrest in developmental time, corresponding to the period of cold treatment.

Pupae storage effect of Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) in low temperature.

The study was developed in the 'Laboratorio de Biologia e Criacao de Insetos' (LBCI), 'Faculdade de Ciencias Agrarias e Veterinarias (FCAV), 'Universidade Estadual Paulista' (Unesp), Jaboticabal, SP, with objective of evaluating the pupae storage effect of Cotesia flavipes in low temperature (medium temperature = 2,75[degree]C). The experiment was made up of 13 treatments: fi, 1, 2, 3, 4, 5, 6, 7, 8, 9.10 and 11 masses storage days of pupae in refrigerator temperature and the check was man maintained on 25[plus or minus]2[degree]C and relative humidity of 70[plus or minus]10%. The next generation of parasitoid also was analyzed, maintained to 25[plus or minus]2[degree]C during all development. It was evaluated: period and pupae viability; average number, longevity and adults' sex ratio of the stored generation and period egg-larvae; period and pupae viability: average number, longevity and adults' sex ratio of the next generation. It verified that the pupae storage of C. flavipes in refrigerator temperature for until 5 days does not affect its development and the stored generation, suggesting the utilization possibility of this resource in mass rearing of this insect.

Ultrastructural and Cytochemical Aspects of Metamorphosis in the Midgut of Apis mellifera L. (Hymenoptera: Apidae: Apinae)

The midgut of Apis mellifera is remodeled during metamorphosis. The epithelium and, to a lesser extent, the muscular sheath degenerate between the end of the last larval instar and the onset of pupation (prepupa).The larval epithelium is shed to the midgut lumen and digested, while a new epithelium is reconstructed from larval regenerative cells. During pupation, some reorganization still occurs, mainly in brown-eyed pupae. In pharate adult, the midgut wall shows the characteristics of adult, although some cells have pycnotic nuclei. The localization of alkaline and acid phosphatases showed that these enzymes were not involved in the reabsorption of the midgut wall.

Comparison of eggs, larvae, and pupae of plutella xylostella (lepidoptera: Plutellidae) as prey for podisus nigrispinus (Hemiptera: Pentatomidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)