Rearing <Emphasis Type="Italic">Chrysoperla externa</Emphasis> Larvae on Artificial Diets

We tested three artificial diets for rearing larvae of Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae), aiming at reducing the production costs of this predator. Two of the diets come from studies with other species of lacewings, and the third is a modification described in this paper. All diets were based on animal protein and were supplied to 2nd and 3rd instar larvae, whereas 1st instar larvae received eggs of Anagasta kuehniella (Zeller) (Lepidoptera: Pyralidae). We evaluated the preimaginal duration and survival, adult size, longevity and fecundity, egg hatchability, and predatory capacity of larvae produced. The performance of the diets was followed for seven generations. The diet we describe showed to be the best among the artificial diets tested. Our results show that C. externa can be successfully reared on artificial diets during second and third instars, reducing in 90% the dependency on eggs of A. kuehniella.     

Interspecific competition among endoparasitoids of tobacco budworm larvae [Lep.: Noctuidae]

The ability of female larvae endoparasitoids [Microplitis croceipes Cresson:Cardiochiles nigriceps Viereck andCumpoletis sonorensis (Carlson)] to distinguish between unparasitized tobacco budworm,Heliothis virescens (F.), larvae andH. virescens larvae parasitized by the egg-larval parasitoidChelonus insularis,Cresson, was determined in laboratory studies. The 3 species of larval endoparasitoid females did not appear capable of distinguishing between unparasitized andC. insularis parasitized larvae resulting in multiple parasitoidism. The results of the ensuing competition between the 3 species for possession of the host demonstrated that bothC. sonorensis andM. croceipes were intrinsically superior toC. insularis. Both larva endoparasitoids destroyed the olderC. insularis larvae by physically attacking the latter. The presence ofC. insularis larvae in the host was found to prevent the hatch of compeatingC. nigriceps eggs through physiological suppression. The results show that the early attack of a host, as in the egg-larval parasitoid habit, is not necessarily advantageous.     

Larvae of phoronids (Phoronida) from Terpeniya Bay (Southeastern Sakhalin)

Phoronid larvae were found in planktonic samples from the northern coast of Terpeniya Bay. In some samples, their density was up to 220 specimens/m³. Larval stages having 10, 12, 16, 20, and 22 tentacles are described. Larvae were identified as Actinotrocha branchiata and belong to the species Phoronis muelleri Selys-Longchamps, 1903. However, unlike the Ph. muelleri larvae described in the literature, the larvae we found are smaller (not more than 900 μm) before metamorphosis and have fewer tentacles (24). They lack paired vacuolated diverticula of the stomach, which are characteristic of Ph. muelleri larvae. However, judging by all other characters, namely transparency, the absence of coelomic cylinder in the preoral lobe, and the presence of adult tentacle primordia, one pair of blood cell aggregations, and a pyriform organ, these larvae are similar to the previously described larvae of Ph. muelleri. Adult forms of Ph. muelleri were previously found in Terpeniya Bay and described by Mamkaev (1962) and Emig (1984).     

A description of the larvae of <Emphasis Type="Italic">Cinygmula unicolorata</Emphasis> Tshernova, 1979 and <Emphasis Type="Italic">Cinygmula malaisei</Emphasis> (Ulmer, 1927) (Ephemeroptera,Heptageniidae) from the Russian Far East

The larvae of Cinygmula malaisei (Ulmer, 1927) and Cinygmula unicolorata Tshernova, 1979 from the Russian Far East are described and illustrated. The larva of Cinygmula unicolorata is similar to those of Cinygmula putoranica Kluge, 1980 and Cinygmula uyka Gorovaya et Tiunova, 2013 but differs from the larva of C. uyka in the absence of gill filaments and from that of C. unicolorata in the shape of the first gill (tergalia). The upper outer margin of the first gill is rounded in the larva of C. unicolorata and almost straight in C. putoranica; the gills of C. unicolorata are matte and the trachea is poorly expressed, in contrast to the distinctly brown trachea of C. putoranica. The larvae of Cinygmula malaisei are similar to those of C. irina and C. autumnalis but differ from the latter in the presence of gill filaments. The larva of C. malaise differs from that of C. irina in the rounded outer margin of the first gill, the shape of gills VI and VII, and in the size of the leg segments.     

The oxygen consumption of mayfly (Ephemeroptera) and stonefly (Plecoptera) larvae at different oxygen concentration

1. The aim of this investigation was to elucidate how four acquatic insect larvae, from different habitats and having different respiratory organs or types of respiratory regulation, react to a lowered oxygen concentration, and how their oxygen consumption is affected. The species investigated were the stoneflies Taeniopteryx nebulosa, Diura nanseni and Nemoura cincerea and the mayfly Cloëon dipterum.
2. The measurements were performed in a respiratory apparatus of open, flowing-water type. Its design is shown in Fig. 1. Water of known oxygen concentration was allowed to flow past the experimental larvae. The oxygen consumption of the larvae was calculated from the lowering of the oxygen concentration in which ensued.
3. The water used in the experiments was standardized, so that the electrode had the necessary stability (conductivity 470 micromhos/cm). The calcium ion was excluded in order to prevent the precipitation of CaCO₃ in the electrode capillary.
4. A large variation in the values of oxygen consumption was found as seen in Fig. 2–5. The reason for that is a corresponding variation in the motor activity of the experimental animals.
5. The physiological reasons for the general form of the curves A and C in Fig. 2–5 are discussed. The curves A and C represent oxygen consumption of the larvae at different degrees of stimulation, entailing different levels of motor activity. Curve A represents intentinally activated animals, curve C non-activated, motionless animals. The curves A and C are boundary curves corresponding to a sort of scope for activity of the animals. Over this scope area a series of curves of the same form could in principal be construed, representing different degrees of stimulation.
6. Within a certain oxygen concentration interval a motor activation was observed caused by a reduced oxygen concentration. The result of that activation can be seen in Fig. 2–5 as a zone with no or very few oxygen consumption values between curve C and D. The more easily activated the species is, the broader the zone will be. Cloën has the most narrow zone and was observed to be less activated than the other species.
7. Small larvae of Cloën (2–4 mm and 42–6 mm) and Nemoura (2–4 mm) showed clearly a greater ability to take up oxygen at low oxygen concentrations than full-grown larvae (see Fig. 8 and 9).
8. The critical point on the curve representing mean oxygen consumption as a function of oxygen concentrations was found to be at 2–5 mg O₂/1 for Taeniopteryx and Diura, at 2.2–2.5 mg O₂/1 for Cloëon, and at about 2–7 mg O₂/1 for Nemoura. The values refer to 8°. Cloëon is the species which is exposed to the greatest variations in oxygen concentration in its natural environment.
9. No influence on the oxygen consumption of starvation for 4 to 5 days was found. No difference between the oxygen consumption values obtained in the presence or in the absence of calcium ions could be observed during the experiments (Fig. 10, 11).
10. The basic picture obtained in this investigation is a set of oxygen consumption values scattered between a curve connecting highest values obtained and a curve of the standard metabolism together with a zone in which the larvae are activated by reduced oxygen concentrations. This picture is presumed to be general in aquatic animals with a well developed motor activity.

     

The effect of temperature on the swimming of a teleost (Clupea harengus) and an ascidian larva (Dendrodoa grossularia)

• 1.1. Using a high-speed video system operating at 400 frames/sec, the effects of temperature on tail beat frequency, swimming speed and stride length were examined in newly hatched larvae of herring (Clupea harengus L.) and in tadpole larvae of the ascidian Dendrodoa grossularia van Beneden.
• 2.2. The effect of temperature was linear; the tail beat frequency of 8 mm-long herring larvae increased from 19 Hz at 5.6°C to 37 Hz at 14.9°C (Q₁₀ = 2.04); that of 2 mm-long Dendrodoa larvae increased from 10 Hz at 9.6°C to 23 Hz at 18.1°C (Q₁₀ = 2.52).
• 3.3. Burst swimming speeds of herring larvae increased from 80 mm/sec at 5°C to 150 mm/sec at 15°C, stride length remaining constant at about 0.5 of the body length for each tail beat.
• 4.4. More continuous swimming of Dendrodoa increased from 4.0 mm/sec at 10°C to 11.5 mm/sec at 18°C, the stride length increasing from about 0.15 to 0.25.

     

Reproduction and larval development of the gastropod <Emphasis Type="Italic">Cryptonatica janthostoma</Emphasis> (Gastropoda: Naticidae)

The larval morphology of the gastropod Cryptonatica janthostoma inhabiting the Northwest Pacific was described for the first time. Hatched planktotrophic veligers of C. janthostoma had shells 250 μm in height with 0.8 whorls, a bilobate velum with a dark brown pigmentation band shaped along its edge, pair of eye spots, tentacles and statocysts. The surface of the embryonal shell (protoconch 1) was covered with fine granules extended at the dorsal side and rounded at lateral surfaces. Concentric crests and growth lines occurred on part of the shell of late free-swimming larvae (protoconch 2) The velum of the C. janthostoma larvae remained bilobate during the pelagic stage of development, whereas it was divided in 4 lobes during larval development in most of the other species of the Naticidae family. The veliger of C. janthostoma was similar to that of Natica montagu from Danish waters [16] by its shape, pigmentation, shell sculpture and velum structure.     

A New Species of the Genus <Emphasis Type="Italic">Placoblatta</Emphasis> Bey-Bienko (Dictyoptera,Blaberidae, Epilamprinae) from Southern Vietnam

A new species of the genus Placoblatta, P. minorsp. n., is described from Southern Vietnam. The similarity between representatives of the genus Placoblatta and larvae of the subfamily Epilamprinae is briefly discussed. It is suggested that the origin of the genus Placoblatta has a paedomorph nature.     

Novel, meso-substituted cationic porphyrin molecule for photo-mediated larval control of the dengue vector Aedes aegypti

Background

Control of the mosquito vector population is the most effective strategy currently available for the prevention of dengue fever and the containment of outbreaks. Photo-activated oxidants may represent promising tools for developing effective, safe and ecofriendly novel larvicides. The purpose of this study was to evaluate the potential of the synthetic meso-substituted porphyrin meso-tri(N-methylpyridyl), meso-mono(N-tetradecylpyridyl)porphine (C14) as a photoactivatable larvicide against the dengue vector Aedes (Stegomyia) aegypti.

Methodology

The photophysical and photochemical properties of the C14 molecule were assessed spectrophotometrically. Photomediated larvicidal efficacy, route of intake and site of action were determined on Ae. aegypti larvae by laboratory bioassays and fluorescence microscopy. Using powdered food pellet for laboratory rodents (a common larval food used in the laboratory) as a carrier for C14, loading-release dynamics, larvicidal efficacy and residual activity of the C14-carrier complex were investigated.

Main Findings

The C14 molecule was found to exert a potent photosensitizing activity on Ae. aegypti larvae. At irradiation intervals of 12 h and 1 h, at a light intensity of 4.0 mW/cm², which is 50–100 times lower than that of natural sunlight, LC₅₀ values of 0.1 µM (0.15 mg/l) and 0.5 µM (0.77 mg/l) were obtained, respectively. The molecule was active after ingestion by the larvae and caused irreversible, lethal damage to the midgut and caecal epithelia. The amphiphilic nature of C14 allowed a formulate to be produced that not only was as active against the larvae as C14 in solution, but also possessed a residual activity of at least two weeks, in laboratory conditions.

Conclusions

The meso-substituted synthetic porphyrin C14, thanks to its photo-sensitizing properties represents an attractive candidate for the development of novel photolarvicides for dengue vector control.     

Patterns of microbial food webs in Mediterranean shallow lakes with contrasting nutrient levels and predation pressures

Hygraula nitens is a New Zealand native moth with aquatic larvae that feed on submerged aquatic plants. The larvae have been mainly observed using native Potamogeton and Myriophyllum species as a food source, although some studies reported larvae feeding on the alien macrophytes Hydrilla verticillata, Lagarosiphon major and Ceratophyllum demersum. Experimental mesocosm studies showed larvae had a major effect on H. verticillata, C. demersum, L. major, Elodea canadensis and Egeria densa. In both no choice and choice experiments H. nitens larvae showed a clear preference for and the highest consumption of C. demersum, while the native macrophyte Myriophyllum triphyllum ranked fourth out of five alien and two native plant species, indicating a preference of the larvae for alien macrophytes. Additional choice experiments using C. demersum, sampled from different waters in NZ, illustrated that there was a clear difference in H. nitens preference for plants based on their source. However although C. demersum had the lowest leaf dry matter content (LDMC) compared with the other macrophytes, neither the LDMC nor leaf carbon, nitrogen, phosphorus or total phenolic contents alone could explain the preferences of H. nitens, and we conclude that food choice is based on a combination of these and/or additional factors.     

The Function of a Spindle Checkpoint Gene bub-1 in C. elegans Development

Background

The serine/threonine kinase BUB1 (Budding Uninhibited by Benzimidazole 1) was originally identified in yeast as a checkpoint protein, based on its mutant''s incapacity of delaying the cell cycle in response to loss of microtubules. Our understanding of its function is primarily from studies carried out in yeast S. cerevisiae. It has been shown that it is a component of the mitotic spindle checkpoint and regulates the separation of sister chromatids through its downstream molecules. However, its roles in multi-cellular organisms remain unclear.

Methods and Findings

In nematode C. elegans, rapid cell divisions primarily occur in embryos and in germline of postembryonic larvae and adults. In addition, a select set of cells undergo a few rounds of cell division postembryonically. One common phenotype associated with impaired cell division is described as Stu (Sterile and Uncoordinated) [1], [2]. We conducted a genetic screen for zygotic mutants that displayed Stu phenotype in C. elegans. We isolated seven Stu mutants that fell into five complementation groups. We report here that two mutations, FanWang5 (fw5) and FanWang8 (fw8) affect the bub-1 gene, a homolog of yeast BUB1. Both mutant alleles of fw5 and fw8 exhibited variable behavioral defects, including developmental arrest, uncoordination and sterility. The number of postembryonically born neurons in the ventral cord decreased and their axon morphology was abnormal. Also, the decrease of neurons in the ventral cord phenotype could not be suppressed by a caspase-3 loss-of-function mutant. In addition, bub-1(fw5 and fw8) mutants showed widespread effects on postembryonic development in many cell lineages. We found that bub-1 functioned maternally in several developmental lineages at the embryonic stage in C. elegans. Studies in yeast have shown that BUB1 functions as a spindle checkpoint protein by regulating the anaphase promoting complex/cyclosome (APC/C). We performed double mutant analysis and observed that bub-1 genetically interacted with several downstream genes, including fzy-1/CDC20, mat-2/APC1 and emb-27/APC6.

Conclusions

Our results demonstrate a conserved role of bub-1 in cell-cycle regulation and reveal that C. elegans bub-1 is required both maternally and zygotically. Further, our genetic analysis is consistent with that the function of bub-1 in C. elegans is likely similar to its yeast and mammalian homologs.     

Offspring dependence on parental care and the role of parental transfer of oral fluids in burying beetles

Background

Immature stages of many animals can forage and feed on their own, whereas others depend on their parents’ assistance to obtain or process food. But how does such dependency evolve, and which offspring and parental traits are involved? Burying beetles (Nicrophorus) provide extensive biparental care, including food provisioning to their offspring. Interestingly, there is substantial variation in the reliance of offspring on post-hatching care among species. Here, we examine the proximate mechanisms underlying offspring dependence, focusing on the larvae of N. orbicollis, which are not able to survive in the absence of parents. We specifically asked whether the high offspring dependence is caused by (1) a low starvation tolerance, (2) a low ability to self-feed or (3) the need to obtain parental oral fluids. Finally, we determined how much care (i.e. duration of care) they require to be able to survive.

Results

We demonstrate that N. orbicollis larvae are not characterized by a lower starvation tolerance than larvae of the more independent species. Hatchlings of N. orbicollis are generally able to self-feed, but the efficiency depends on the kind of food presented and differs from the more independent species. Further, we show that even when providing highly dependent N. orbicollis larvae with easy ingestible liquefied mice carrion, only few of them survived to pupation. However, adding parental oral fluids significantly increased their survival rate. Finally, we demonstrate that survival and growth of dependent N. orbicollis larvae is increased greatly by only a few hours of parental care.

Conclusions

Considering the fact that larvae of other burying beetle species are able to survive in the absence of care, the high dependence of N. orbicollis larvae is puzzling. Even though they have not lost the ability to self-feed, an easily digestible, liquefied carrion meal is not sufficient to ensure their survival. However, our results indicate that the transfer of parental oral fluids is an essential component of care. In the majority of mammals, offspring rely on the exchange of fluids (i.e. milk) to survive, and our findings suggest that even in subsocial insects, such as burying beetles, parental fluids can significantly affect offspring survival.

     

Gut Bacteria Are Not Required for the Insecticidal Activity of Bacillus thuringiensis toward the Tobacco Hornworm,Manduca sexta

It was recently proposed that gut bacteria are required for the insecticidal activity of the Bacillus thuringiensis-based insecticide, DiPel, toward the lepidopterans Manduca sexta, Pieris rapae, Vanessa cardui, and Lymantria dispar. Using a similar methodology, it was found that gut bacteria were not required for the toxicity of DiPel or Cry1Ac or for the synergism of an otherwise sublethal concentration of Cry1Ac toward M. sexta. The toxicities of DiPel and of B. thuringiensis HD73 Cry⁻ spore/Cry1Ac synergism were attenuated by continuously exposing larvae to antibiotics before bioassays. Attenuation could be eliminated by exposing larvae to antibiotics only during the first instar without altering larval sterility. Prior antibiotic exposure did not attenuate Cry1Ac toxicity. The presence of enterococci in larval guts slowed mortality resulting from DiPel exposure and halved Cry1Ac toxicity but had little effect on B. thuringiensis HD73 Cry⁻ spore/Cry1Ac synergism. B. thuringiensis Cry⁻ cells killed larvae after intrahemocoelic inoculation of M. sexta, Galleria mellonella, and Spodoptera litura and grew rapidly in plasma from M. sexta, S. litura, and Tenebrio molitor. These findings suggest that gut bacteria are not required for B. thuringiensis insecticidal activity toward M. sexta but that B. thuringiensis lethality is reduced in larvae that are continuously exposed to antibiotics before bioassay.Bacillus thuringiensis has long been regarded as a bona fide entomopathogen that can produce an array of virulence factors including insecticidal parasporal crystal (Cry) toxins, vegetative insecticidal proteins, phospholipases, immune inhibitors, and antibiotics (31). B. thuringiensis establishes lethal infections in many insect species after intrahemocoelic inoculation (9, 10, 14, 26, 31), and the insecticidal activity of Cry toxins, which lyse the intestinal epithelium, can be synergized by the presence of viable B. thuringiensis spores (31). In each instance, synergism has been attributed to hemocoelic infection by B. thuringiensis.A novel hypothesis (6, 7) proposed that B. thuringiensis is incapable of killing Lymantria dispar, Manduca sexta, Pieris rapae, or Vanessa cardui in the absence of gut bacteria. Prior exposure of L. dispar larvae to a combination of four antibiotics severely reduced the subsequent toxicity of the B. thuringiensis-based (spores and Cry toxins) bioinsecticide, DiPel (Valent BioSciences) (6). Both larval susceptibility to B. thuringiensis and the number of culturable gut bacteria were found to be negatively correlated with the concentration of antibiotics to which larvae were previously exposed. Furthermore, a total reduction in larval susceptibility was coincident with the elimination of any detectable gut bacteria. Experimental reinfection with Enterobacter sp. strain NAB3, found in the guts of some populations of L. dispar larvae, was found to rescue the toxicity of B. thuringiensis, whereas reinfection with Enterococcus casseliflavus and Staphylococcus xylosus did not. It was also shown that while Escherichia coli, Enterobacter sp. strain NAB3, and B. thuringiensis could all grow in tryptic soy broth, B. thuringiensis alone could not grow in filter-sterilized plasma from L. dispar larvae. Finally, it was shown that the toxicity of Cry1Aa-expressing E. coli JM103 to L. dispar larvae was reduced by the prior exposure of larvae to antibiotics and could be eliminated when E. coli was also heat killed before use. It was concluded that B. thuringiensis-induced mortality results from a mixed infection of the hemocoel that must include bacteria capable of growth within the L. dispar larval hemolymph (6).Using the same methods, it was subsequently reported that prior exposure of Vanessa cardui, M. sexta, Pieris rapae, and Heliothis virescens larvae to antibiotics eliminated culturable bacteria and rendered larvae resistant to DiPel (7). Experimental reinfection of larvae with Enterobacter sp. strain NAB3 rescued DiPel toxicity in V. cardui, M. sexta, and P. rapae but not in H. virescens larvae. Using a continuous-exposure bioassay, the susceptibility of Pectinophora gossypiella to the Cry1Ac-based bioinsecticide MVPII was found to be increased by prior exposure to antibiotics. Toxicity from a 48-h exposure of L. dispar larvae to MVPII was reduced, but not eliminated, by prior antibiotic exposure and could be rescued by reinfection with Enterobacter sp. strain NAB3. It was concluded that “enteric bacteria have important roles in B. thuringiensis-induced killing of Lepidoptera across a range of taxonomy, feeding breadth, and relative susceptibility to B. thuringiensis” (7).The present work shows that gut bacteria are not required for the insecticidal activity of B. thuringiensis or Cry1Ac toxin toward M. sexta but that prior antibiotic exposure reduces larval susceptibility to B. thuringiensis.     

Bt Proteins Have No Detrimental Effects on Larvae of the Green Lacewing, <Emphasis Type="Italic">Chrysopa pallens</Emphasis> (Rambur) (Neuroptera: Chrysopidae)

Biosafety of a genetically modified crop is required to be assessed prior to its commercialization. For this, a suitable artificial diet was developed and used to establish a dietary exposure test for assessing the toxicity of midgut-active Bt insecticidal proteins on Chrysopa pallens (Rambur). Subsequently, this dietary exposure test was used to evaluate the toxicity of the proteins Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa on C. pallens larvae. Temporal stability, bioactivity, and the intake of the insecticidal proteins were confirmed by enzyme-linked immunosorbent assay and a sensitive-insect bioassay. The life history characteristics, such as survival, pupation, adult emergence, 7-day larval weight, larval developmental time, and emerged male and female fresh weights remained unaffected, when C. pallens were fed the pure artificial diet (negative control) and the artificial diets containing 200 μg/g of each purified protein: Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, or Vip3Aa. On the contrary, all of the life history characteristics of C. pallens larvae were adversely affected when fed artificial diet containing boric acid (positive control). The results demonstrate that diets containing the tested concentrations of Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa have null effects on C. pallens larvae. The outcome indicates that genetically modified crops expressing the tested Bt proteins are safe for the lacewing, C. pallens.     

A morphological and phylogenetic analysis of <Emphasis Type="Italic">Ornithodoros marinkellei</Emphasis> (Acari: Argasidae), with additional notes on habitat and host usage

Ornithodoros marinkellei was described from larvae collected on Pteronotus spp. bats in Colombia and Panama. More recently, this tick was reported in the Brazilian Amazon. Because some morphometric differences were observed between O. marinkellei larvae from Colombia and Brazil, it was proposed that further investigations were needed to assess whether the differences could be attributed to intra- or inter-specific polymorphism. Herein, we collected O. marinkellei specimens in the type locality of Colombia, in Brazil, and in a new locality in Nicaragua, expanding the distribution of the species to Nicaragua. Morphometric analysis of larvae and adults, corroborated by a principal component analysis (PCA), indicated that the Brazilian specimens were larger than specimens from Colombia and Nicaragua. Phylogenetic analysis inferred from the mitochondrial 16S rRNA gene showed ticks from Colombia and Nicaragua more genetically related than any of them with ticks from Brazil, although ticks from the three countries grouped in a clade sister to a major clade containing sequences of various Neotropical Ornithodoros species. We concluded that ticks identified as O. marinkellei from Colombia, Nicaragua, and Brazil represent the same taxon, and that the genetic and morphological differences between them are likely to have a geographical bias. We redescribed the nymph of O. marinkellei, which has a vestigial hypostome, probably incompatible with blood feeding. We also report human infestation by O. marinkellei adults. As all reports of O. marinkellei adults have been from hot caves (temperature?>?35 °C), this abiotic condition could be a limiting factor for the occurrence of this tick species.     

Effect of low rearing temperature on development of Galleria mellonella larvae and their sensitivity to juvenilizing treatment

• 1.1. The development of Gallena mellonella is strongly affected by a low temperature of 18°C (the last instar persists for more than one year, instead of about 9 days at 30°C). At 18°C the last instar Galleria mellonella larvae respond to juvenilizing treatment—chilling stress or juvenile hormone analogue—with a very low percentage or no supernumerary moults, respectively.
• 2.3. Experiments in which larvae subjected to such treatments were transferred from 18°C to 30°C and vice versa showed that for the realization of the larval programme after chilling stress application the higher (30°C) temperature is needed.
• 3.4. In last instar larvae reared at 18°C there coexist very high juvenile hormone titre and high juvenile hormone esterase activity.
• 4.5. This phenomenon which is found in both, chilled and unchilled larvae, is discussed.