Effects of a juvenile hormone mimetic,fenoxycarb, on post-embryonic development of the European corn borer,Ostrinia nubilalis Hbn.

Summary The effect of a juvenile hormone mimetic, fenoxycarb, Ro 13-5223, was tested on the larval instars of the European corn borer,Ostrinia nubilalis, by dipping or topical application. When larvae were treated in instars 2, 3 or 4, the duration of the fifth instar was modified. More permanent and fewer supernumerary larvae were obtained when treatment occurred in the early instars. This non-neurotoxic compound exhibited a strong dose-dependent juvenile hormone type of activity when it was applied to last instar larvae. Fenoxycarb prevented the onset of pupation and produced supernumerary larvae and intermediates. Permanent larvae were obtained if fenoxycarb was applied on day 0 or day 1 of the last instar. The use of such a JH mimetic in the understanding of endocrine control of diapause is discussed.     

The influence of juvenile hormone on the feeding behaviour of last instar larvae of the codling moth,Laspeyresia pomonella (Lep., Tortricidae), reared under different photoperiods

Summary Duration of the feeding stage and corresponding weight increase during the last larval instar of the codling moth,Laspeyresia pomonella, are controlled by JH. Larvae reared under short day conditions have a relatively high titer of JH during the last larval instar and enter diapause as mature larvae. They feed longer and become heavier than larvae reared under long day conditions, which have no JH during the last larval instar and pupate when mature. By application of the JH mimetic Altosid^® during the first 2 or 3 days of the last larval instar, the duration of feeding activity of larvae reared under respectively long and short day conditions was prolongated and the larvae became significantly heavier. The feeding behaviour could only be influenced by the juvenoid as long as the feeding activity of the larvae had not yet ceased.     

Head critical period and juvenile hormone titre during the last larval instar of the cockroach,Periplaneta americana

From neck ligation experiments with last instar larvae of the cockroachPeriplaneta americana it was concluded that the head critical period is reached around day 17, which corresponds to 59% of the last larval stage. At the same stage the juvenile hormone III titre in the hemolymph dropped to undectable levels.     

Juvenilizing effect of ecdysone mimic RH 5849 inGalleria mellonella larvae

The response of the final instar larvae ofG. mellonella to topical application of the non-steroidal ecdysone mimic, RH 5849, was age-related as well as dose-dependent. In young final instar larvae, moderate doses of RH 5849 induced perfect supernumerary larval moults, but doses equal to and higher than 8.5 μg per larva caused premature formation of larval cuticle and were lethal. Application of RH 5849 significantly increased allatotropic activity of the brain, and also activated synthesis of juvenile hormone (JH) by the corpora cardiaca/corpora allata complex. Simultaneous application of RH 5849 and FMev, a potent inhibitor of JH synthesis, to young final instar larvae lowered the incidence of perfect supernumerary larval moults. We conclude that the effect of RH 5849 on the developmental programme inG. mellonella is mediated by the corpora allata.     

Analysis of the relationship between age of larvae at mutagen treatment and frequency and size of spots in the wing somatic mutation and recombination test inDrosophila melanogaster

The relationship between the induction of mutant clones and the time of mutagen treatment was studied in the somatic mutation and recombination test (SMART) in wing cells ofDrosophila melanogaster. Larvae trans-heterozygous for the recessive marker mutations multiple wing hairs (mwh) and flare (flr) were produced. Batches of these larvae were then treated with mutagen at different ages spanning all three larval instars. Methyl methanesulfonate was fed acutely for 2 h by immersing the larvae in a solution of the mutagen mixed with powdered cellulose. Wings of the surviving adult flies were mounted and scored for the presence of spots. The frequency and size of single and twin spots were recorded separately. Twin spots are produced exclusively by mitotic recombination, whereas single spots can results from various types of mutational and exchange events. There exists a clear correlation between time of induction and frequency as well as size of the single spots. In young larvae only few but very large spots are induced, whereas in older larvae the frequencies are considerably increased but the sizes are smaller. The twin spots show a different relationship. Practically no twin spots are found in very young and in very old larvae. The results demonstrate that in the wing spot test the optimal age of the larvae for mutagen treatment is 72 h.     

Role of low ecdysteroid levels in the early last larval instar ofBombyx mori

The relationship between juvenile hormone and ecdysteroid titers during the early stage of the last larval instar of the silkworm,Bombyx mori, was examined in this study. When larvae were fed 20-hydroxyecdysone-supplemented mulberry leaves throughout the last larval instar, 100% underwent supernumerary larval molting instead of metamorphosis. The application of juvenile hormone mimic during the early last instar did not induce supernumerary larval molting, but did delay metamorphosis. Temporal and quantitative ecdysteroid titer measurements revealed that in normal larvae the titers maintained very low levels (3–12 ng/ml) during the early stage of the last instar; however, in 20-hydroxyecdysone-treated larvae, levels ranging from 24 to 45 ng/ml were detected, and a major peak (246 ng/ml) was observed on day 6. These results show that very low ecdysteroid titer levels during the early stage of the last larval instar may play an important role in initiating decreases in juvenile hormone titers as well as in directing metamorphosis.     

Cannibalism inAnopheles pharoensis Theo.

Summary A single 4th instar larva ofAnopheles pharoensis could consume within 24 h an average number of 5.3–11.6 larvae of the 1st instar. The number consumed differed according to crowdedness and the presence of other mature larvae. Dissection of the midgut of these larvae revealed the presence of various undigested parts of young larvae.Acknowledgments. This work was carried out in the Institute of Genetics, Mainz University, Federal Republic of Germany, through a fellowship to the author by the Alexander von Humbolt Foundation, Bonn, FRG, to whom the author is thankful for their support.     

The clerid beetle,Thanasimus formicarius,is attracted to the pheromone of the ambrosia beetle,Trypodendron lineatum

Summary Sticky traps containing (+)-lineatin, the pheromone of the ambrosia beetle,Trypodendron lineatum, attracted the predatorThanasimus formicarius to about the same extent as traps baited with ipslure, the pheromone blend used for mass-trappingIps typographus. The results indicate thatT. lineatum is an important prey forT. formicarius early in the season before the main prey becomes active. Addition ofexo-brevicomin to ipslure and ethanol and/or -pinene to (+)-lineatin did not significantly influence the catches of the predator.     

An empirical relationship between the prey density and predatory efficiency ofGambusia affinis

Summary Larval density of Culex fatigans essentially influences the predatory rate of the mosquito-fish Gambusia affinis. The feeding rate of the fish gradually increases up to a maximum value at intermediate prey densities beyond which there is an asymptotic saturation in the feeding rate. The relationship is represented by an empirical equation which helps in the prediction of the ideal stocking rate of the fish in natural systems.     

Brain factor fromGalleria mellonella (Lepidoptera) stimulating silk gland activity

Brain extracts from day 1–4 last instar larvae ofGalleria mellonella (Lepidoptera) stimulate RNA synthesis in cultured silk glands from day 3 last instar larvae. When the fibroin-synthesizing posterior parts of silk glands were incubated for 3 h in vitro in the presence of brain extract (0.1 brain equivalent), [³H]-uridine incorporation into RNA was stimulated more than twofold. The stimulating effect of brain extract showed a dose response relationship. It is suggested that the heat-resistant and protease-sensitive brain factor is a peptide.     

Juvenile hormone titers in penultimate and last instar larvae ofPieris brassicae andBarathra brassicae,in relation to the effect of juvenoid application

Summary JH titer was determined in the haemolymph of penultimate and last instar larvae ofPieris brassicae L. andBarathra brassicae L. The differences we observed were consistent with physiological differences between the two species.The authors feel indebted to Prof.V. J. Brookes for useful comments.     

Growth,development, feeding preference,and food consumption and utilization by tobacco hornworm on tobacco mosaic virus-infected and non-infected tobacco leaves

Summary Plant disease may cause a variety of changes in plants which influence their suitability to insect herbivores. The reported physiological changes in tobacco due to infection by tobacco mosaic virus have the potential to influence the feeding, growth and behavior of the tobacco hornworm,Manduca sexta. However, feeding on foliage of systematically infected plants had no statistically significant effects on development time, mean pupal weight or the amount of foliage consumed by 4th instars. Although an initial preference was exhibited by larvae for healthy leaves after about 72 h no differences were observed. These results do not support the results of previous research on the influence of tobacco mosaic virus-infected plants on the tobacco hornworm.Scientific contribution No. 8130, article No. A-5071, of the Maryland Agricultural Experiment Station, Department of Entomology     

Expression and immunolocalisation of odorant-binding and chemosensory proteins in locusts

We have identified, cloned and expressed a new chemosensory protein (CSP) in the desert locust Schistocerca gregaria belonging to a third sub-class of these polypeptides. Polyclonal antibodies stained a band of 14 kDa, as expected, in the extracts of antennae and palps of the adults, but not in the 4th and 5th instars. In the related species Locusta migratoria, instead, the same antibodies cross-reacted only with a band of apparent molecular mass of 35 kDa in the extract of 1st–5th instars, but not in the adults. The recombinant protein binds the fluorescent probe N-phenyl-1-naphthylamine, but none of the compounds so far reported as pheromones for S. gregaria. The expression of the odorant-binding protein (OBP) and of CSPs of sub-classes I and II was also monitored in antennae, tarsi, palpi, wings and other organs of solitary and gregarious locusts in their nymphal and adult stages. OBP was found to be antenna specific, where it is expressed at least from the 3rd instar in both solitary and gregarious locusts. CSPs, instead, appear to be more ubiquitous, with different expression patterns, according to the sub-class. Immunocytochemistry experiments revealed that OBP is present in the sensillum lymph of sensilla trichodea and basiconica, while CSP-I and CSP-III were found in the outer sensillum lymph of sensilla chaetica and in the sub-cuticular space between epidermis and cuticle of the antenna. Sensilla chaetica on other parts of the body showed the same expression of CSP-I as those on the antenna.Received 11 Janury 2005; received after revision 21 February 2005; accepted 18 March 2005X. Jin and A. Brandazza contributed equally to this work.     

Orientation ofPapilio demoleus larvae to coloured solutions

Summary The present work compares the taxis ofPapilio demoleus larvae to different coloured solutions. The larval positive taxis, i.e., attraction, is maximum for yellow colour and declines with the increase or decrease in the wavelength of maximum light transmission. Red and bluish-green colours repel the larvae.     

Spermatogenesis is extraordinarily accelerated in metamorphosis-arrested larvae of a salamander,Hynobius retardatus

Laboratory experiments were conducted to induce neoteny inHynobius retardatus, which had been reported to propagate in larval forms like axolotl. A large number of newly hatched larvae were reared in an aqueous solution of thiourea (TU) and sodium perchlorate (SPC) in order to arrest the metamorphosis. Gonadal development in the metamophosis-arrested larvae was compared with that in normally metamorphosing and metamorphosed controls. Metamorphosis-arrested male larvae produced morphologically mature spermatozoa approximately 4 months after hatching, when the gonads in the controls began to differentiate into testes, or to show the premeiotic proliferation of germ cells. Possible endocrine controls of these phenomena are discussed.     

Moult inhibition in the insectDysdercus cingulatus (Insecta: Heteroptera) by the cerebral glands of the millipedeJonespeltis splendidus (Myriapoda,Diplopoda)

Summary Implantation of 2 pairs of cerebral glands of the diplopodJonespeltis into the newly-moulted 4th or 5th instar nymphs of the insectDysdercus delayed the moulting of these insects for a fairly long time. Implantation of cerebral glands into 1-day old 5th instars postponed the subsequent moult for a shorter period, whereas implantation of cerebral glands into 2-day old 5th instars had no effect. These observations suggested that a neurosecretory factor from the cerebral glands inhibited moulting in this insect, and in the case of 5th instar nymphs there was a critical period before which implanted glands were effective in moult inhibition.Acknowledgments. Prof. V.K.K. Prabhu is thanked for suggesting the topic and for guidance. The University of Kerala is thanked for the fellowship awarded. The laboratory facilities afforded are also gratefully acknowledged.     

Ecdysis inhibition inAcrolepiopsis assectella larvae by digitonin: antagonistic effects of cholesterol

Summary Digitonin has a toxic effect onAcrolepiopsis assectella, inhibiting development and molting of young larvae fed on a semi-synthetic diet. This effect depends on concentration. It does not occur when cholesterol is added to the diet.Acknowledgment. We thank Mme Annie Heinzmann for technical assistance.     

The excretion of urea by the larvae ofSpodoptera mauritia boisd. (Noctuidae: Lepidoptera) during development

Summary The concentration of urea in the excreta of the 6th instar larvae ofSpodoptera mauritiavaries from 4.017±0.541 to 31.052±1.193 moles/g dry excreta (mean±SE). The observation confirms that urea excreted is of metabolic origin.     

Detection of toxic metabolites in the hemolymph ofBeauveria bassiana infectedSpodoptera exigua larvae

Highly active metabolites have been detected in the hemolymph of the lepidopteranSpodoptera exigua infected with the mycopathogen,Beauveria bassiana. A combination of phenyl sepharose and CM ion exchange chromatography was utilized to extract the active metabolites from infected hemolymph samples. The active in vivo metabolites, having a molecular mass greater than 10 KDa, were thermolabile and were inactivated by proteinase K. These metabolites were characterized by their ability to disrupt metamorphosis, killing treated larvae at the wandering or pupal stage. Additionally, injection ofS. exigua larvae with active samples caused a reduction in the number of filopodial-producing hemocytes. The biological activities and biochemical properties suggest that novel compounds are produced duringB. bassiana mycosis.