The cocoon spinning behaviour of the chinese oak silkworm, Antheraea pernyi

The cocoon of Antheraea pernyi is constructed in four successive phases, as resolved through movement recordings and time-lapse cinematography and cinefluorography: (1) scaffolding and peduncle (9·2 hr), (2) outer cocoon (13·9 hr), (3) cocoon impregnation (0·7 hr), and (4) inner cocoon (26·9 hr). The caterpillar reverses spinning direction at frequencies characteristic for each phase. The number of cycles (360-degree turns) within a phase is relatively constant from individual to individual, although the length of phase two varies seasonally. During cocoon impregnation the larva executes turns in rapid succession, ensuring the even distribution of a hindgut exudate which coats the cocoon with crystals and speeds tanning of the silk. In the second and fourth phases intracycle behaviour consists of extension-recovery loops of the anterior segments, each followed by a repositioning of the abdomen.     

Cocoon spinning behaviour in Ephestia kuehniella; Correlation with endocrine events

The cocoon spinning behaviour in Ephestia kuehniella Zell. consists of two phases: construction of the outer envelope and spinning of the inner envelope. The first phase can be modified by tactile stimuli received from the surroundings, whereas the second phase is fixed rigidly and probably under the control of the hormones associated with pupation.     

Fat body protein granules and storage proteins in the silkmoth, Hyalophora cecropia

Fat body cells of silkmoth pupae (Hyalophora cecropia ) contain granules, showing a less dense outer zone and a denser, often crystalline, inner portion appear after cocoon spinning and increase until the larval-pupal ecdysis; more granules are formed in females than in males. Urate granules, appearing fibrous in internal structure, first form about the same time, but their accumulation is more gradual, and continues in the pupa. Both types have been isolated by centrifugation. Protein granules dissolve in buffers to yield proteins 1 and 2, with distinct electrophoretic and antigenic properties. These proteins have been isolated individually from pupal fat body extracts by using their different thermal stabilities in phosphate buffer containing MgCl2 and (NH4)2SO4, respectively, and purification was completed by gel chromatography. Protein 1 has a molecular weight of 480,000 and a subunit of 85,000 daltons, while protein 2 gives values of 530,000 and 89,000, respectively. Their amino acid compositions are similar but distinct. Proteins 1 and 2 accumulate in the hemolymph, beginning 3 days before spinning, reach maximal levels at spinning, and then decline in the hemolymph while granules are formed in the fat body, although the total hemolymph protein concentration does not decline at this time. It is concluded that the fat body of the late, feeding larva synthesizes two related "storage proteins" and secretes them in partially crystalline granules as protein reserves for metamorphosis.     

End-products of behaviour versus behavioural characters: a phylogenetic investigation of pupal cocoon construction and form in some North American black flies (Diptera: Simuliidae)

Cocoon spinning was analysed, using video recording and playback, in eighteen Nearctic black fly species, comparing nine Simulium species, six Eusimulium species, Stegopterna mutata, Cnephia dacotensis and Prosimulium mixtum. Fourteen behavioural characters were revealed that produced twenty-two equally parsimonious trees (CI = 0.93, RI = 0.96). Another tree was constructed on the basis of five characters relating to the cocoon structure (end-product characters). The goal of the study was to determine whether characters relating to behavioural components of black fly cocoon spinning or those based on end-products of the behaviour are superior for revealing phylogenetic relationships. This was accomplished by comparing both data sets to a phylogeny constructed with the use of cytological and morphological characters. If taxa are grouped according to end-products (the cocoons) there are some spurious groupings. The behavioural analysis only required one extra step to duplicate the morphological and cytological tree. In the case of black flies, it is more informative to use characters resulting from the analysis of the cocoon spinning behaviour than cocoon morphology.     

Environmental control of larval behaviour and its consequences for ecdysteroid content and pupation in Ephestia kuehniella

ABSTRACT. The behaviour of postfeeding larvae of Ephestia kuehniella Zell. (Lepidoptera, Pyralidae) depends on environmental conditions. Long day and high larval density (LG) stimulate larvae to spin the outer envelope of the pupal cocoon which, in turn, accelerates pupal ecdysis. Constant darkness and isolation in a limited space (DS) inhibit spinning of the outer envelope, and cause extension of the wandering stage and a delay in pupation. This behaviour was also observed in larvae confined in the diet.
In postfeeding LG larvae, an increase in ecdysteroid content coincides with spinning of the outer envelope. In DS larvae, which fail to spin the outer envelope, the ecdysteroid content remains low during the wandering stage, and rises to a prepupal peak after a considerable delay. Wandering larvae inserted into empty outer envelopes showed a premature rise of ecdysteroids, and it is thus concluded that sensory stimulation from the outer envelope is a cue promoting further development.     

The cDNA-structure of the prothoracicotropic hormone (PTTH) of the silkmoth Hyalophora cecropia.

The structure of the prothoracicotropic neurohormone (PTTH) of the silkmoth Hyalophora cecropia was elucidated at the cDNA level. The identified cDNA of 803 nt, which is over 90% identical with the corresponding part of the Samia cynthia ricini PTTH gene, encodes a preprohormone of 240 amino acids. Presence of proteolytic cleavage sites indicates that the preprohormone is split into a signal peptide, an intercalated peptide (64 residues), and the PTTH monomer (125 residues). Preprohormones of H. cecropia, S. c. ricini, Antheraea pernyi, and Bombyx mori diversified considerably in all these parts, indicating that the evolution of PTTH is unusually fast. Since a similarly rapid, and concerted evolution of the corresponding receptor is unlikely, the PTTH activity probably depends on the conservation of relatively few amino acids allowing proper molecular folding.     

First predation record of Canthecona furcellata (Wolff.) (Hemiptera: Pentatomidae) on spinning stage silkworm Antheraea mylitta (Drury)

A new scientific survey elucidates the preferred attack of stink bug Canthecona furcellata (Wolff.) (Hemiptera: Pentatomidae) on the spinning stage of the tropical tasar silkworm, Antheraea mylitta (Drury) (Lepidoptera: Saturniidae). The silkworm A. mylitta produces an excellent quality of wild silk; however, due to predation by C. furcellata, tasar silk production is reduced. The bug C. furcellata is the most invasive larval predator of A. mylitta and predation is high during early instars as well as the molting stage of the larvae. However, for the first time it is reported that the spinning stage is also preferable for attack by the stink bug. Both the nymphs and adults of C. furcellata attack the spinning silkworm; moreover, stink bug attack is observed in groups under field conditions. It is postulated that feeding preference is due to the concealed, non‐movable and less defensive stage of the tasar larvae during spinning. The predation of C. furcellata includes its approach on target larva of the tasar silkworm during spinning, where it inserts the proboscis inside the larval skin through the moist silk network of newly forming or formed cocoon. Most of the spinning larvae die from the attack and the normal seed cocoon fails to form. The mechano‐ and chemoreceptors, present on the antenna and proboscis of C. furcellata, play an important role in prey locating and the feeding mechanism. The life cycle of C. furcellata is also discussed in the present study.     

Performance of polyvoltine silkworm Bombyx mori L. on different mulberry plant varieties

Five cultivated mulberry plant varieties, BSRM‐34, BSRM‐56, BSRM‐57, BSRM‐58 and BSRM‐59, were used to evaluate the effects of plant variety on larval and cocoon characteristics of mulberry silkworm Bombyx mori L. Silkworm larvae undergo their entire larval periods on leaves of mulberry varieties. Different larval characteristics of larval weight, larval length and larval breadth were measured in different growth stages. After rearing, the fifth instar larvae were transferred into a mountage (Chandraki) structure for cocoon rearing for spinning and cocoon production. The cocoons were then harvested and different cocoon characteristics like cocoon weight, shell weight and shell percentage; and other characters like pupal weight and effective rate of rearing were measured. The results showed statistically significant differences among different varieties on larval and cocoon characteristics. The highest performance was observed by feeding the variety BSRM‐34 in respect of the average weight of 10 mature larvae (40.5 g), single mature larval length (7.6 cm) and breadth (1.0 cm), single cocoon weight (1.4 g), shell weight (0.2 g) and pupal weight (1.2 g) while the average performance was recorded by feeding varieties BSRM‐57 and BSRM‐59. The poorest performance was showed by feeding the variety BSRM‐56 (in case of larval characters) and BSRM‐58 (in case of cocoon characteristics). These two varieties also showed the poorest performance for silk yield. Through comparing all the above parameters it was concluded that BSRM‐34 is the best mulberry variety and it is recommended that Bangladeshi farmers adopt that variety for better yield of silk.     

Influence of mulberry cultivars Morus spp. on the production and quality of silkworm Bombyx mori L. (Lepidoptera: Bombycidae) cocoons

The success of sericiculture greatly depends on the availability of mulberry cultivars with agronomic characteristics that maximize the production and quality of silkworm, Bombyx mori L., cocoons. The effect of the cultivars Miura (standard) and Korin, and the hybrids SM 14, SM 63, SK 1, SK 4 and FM 86 was evaluated on silkworm development, and on cocoon production and quality, at the Instituto Agron?mico do Paraná, IAPAR, in Londrina, PR, Brazil (23 +/- 3 degrees C; 61 +/- 8% RH). The experiment was arranged in a randomized block design with four replicates. Larvae feeding either on 'SK 4' or on 'Korin' showed a higher weight compared to that of larvae feeding on 'Miura'. Cocoon weight (both sexes) and pupae weight (male) obtained with larvae fed on SK 4 were higher compared with that of insects that fed on Miura; there was no difference among the remaining treatments. Silk net weight was independent of the cultivar used. Cocoon shell weight was similar amongst insects raised on 'Miura' and on the other mulberry materials. The percentage of cocoon spinning was higher than 90% in all treatments, with the spinning concentrating in the first two days. The proportion of first grade cocoons varied from 80% (SK4) to 90%. The results obtained with the studied cultivars were either similar or superior to that of the standard cultivar Miura with emphasis to 'SK 4', thus showing the potential use of these cultivars in sericulture in the state of Paraná.     

Haemolymph ecdysteroid titres in diapause and nondiapause larvae of the European corn borer, Ostrinia nubilalis

The ecdysteroid titres of last-instar prediapausing, diapausing and nondiapausing larvae of Ostrinia nubilalis were determined by radioimmunoassay. In the nondiapause larvae a major peak of ecdysteroid activity preceded pupation by 24 h and continued through the pupal ecdysis. This peak was correlated with head and thorax critical periods as well as with changes in behaviour and physiology marking the transition from feeding larva to prepupa. Nondiapause larvae also displayed a rise in ecdysteroid titre during the feeding phase of development. This rise was approx one tenth that of the major peak and lasted 32 h. It was not correlated with any overt changes in larval physiology or behaviour. The diapause ecdysteroid profile was distinctive in that the levels measured were all lower than the lowest of the nondiapause curve. No peaks were observed in the diapause titres. Prepupal changes such as spinning and the cessation of feeding were not correlated with any increase in ecdysteroid levels. During diapause termination, under a long-day photoperiod, no increases in ecdysteroid titre were observed for the first 10 days. After 12 days individuals began to show ecdysteroids above the diapause levels. Pupation started after 16 long days.     

Rates of juvenile hormone synthesis control caste differentiation in the stingless bee Scaptotrigona postica depilis

Summary In social insects the expression of caste-specific characters is controlled by juvenile hormone (JH) during definite sensitive periods in preimaginal development. For a number of stingless bee species the existence of such a JH-sensitive period has already been demonstrated. Queen development can be induced by topical JH applications during the cocoon spinning phase of the last larval instar. Neither JH titers nor rates of JH synthesis were known so far for this subfamily of eusocial bees distinguished by a pronounced caste dimorphism. As the pantropically distributed stingless bees with approximately 400 recent species are the largest group of social bees, JH synthesis was studied in one of the species that can be kept under laboratory conditions. An in vitro radiochemical assay was used to measure stage- and caste-specific activities of the corpora allata (CA). For the first time in a eusocial hymenopteran species it was demonstrated how the endocrine system is reacting to trophogenic stimuli capable to induce caste differentiation during larval development. Generally JH synthesis in queen CA was found to be 30–80% higher than in workers during the penultimate and last larval instar, but a strong and distinct caste-specific modulation of JH synthesis was only observed right before the onset of a JH-sensitive period in the cocoon spinning phase of the fifth instar.     

Rate of cocoon production by the larva of the fly Rhynchosciara americana and storage sites of precursors

A chemical analysis of Rhynchosciara americana cocoons at four stages of spinning was performed and the amount and rate of the cocoon production was measured. These results, together with the amount and distribution of the nutrient reserves in the larva during spinning, were used to identify the possible storage sites of cocoon precursors. The physiology, mechanics, and regulation of spinning by R. americana are discussed.     

Distribution of nutrient reserves during spinning in tissues of the larva of the fly, Rhynchosciara americana

The amount of protein, carbohydrate, lipids, and free amino acids were examined in the spinning stage in the fat body, haemolymph, skeletal muscle, and gut of Rhynchosciara americana. Protein and lipids increase in the fat body soon after the animal stopped feeding, probably at the expense of the digestion of the gut contents and of the reserves of the gut wall. Afterwards there is a fall in protein and lipids in the fat body. Haemolymph protein rises a little at the beginning of spinning and then decreases steadily during cocoon production. Carbohydrate and free amino acids decrease from the beginning of spinning in all tissues studied. Quantitatively, the most important decrease of carbohydrate during spinning occurs in the fat body whereas that of free amino acids occurs in the haemolymph. Lipid increases during spinning in the skeletal muscle, probably due to enlargement of the lateral fat body which occurs as a contaminant in the skeletal muscle preparation. The Malpighian tubules contain a large amount of calcium carbonate, which is eliminated during spinning. A correlation of our chemical data with histochemical data recently published is presented and the physiological implications of our findings are discussed in comparison to other insects.     

Tghe tracheation of the wings of adult Lepidoptera

There is much variation in the extent of tracheation of the wings of adult Lepidoptera. This is surveyed. Significantly more tracheal branching is found in species with pupal cocoons. No correlation of tracheation with wing length is apparent. The concentrations of oxygen and carbon dioxide in the air inside cocoons of Antheraea pernyi Guér. (Saturniidae) before emergence are measured. Oxygen supply is probably not limited by the cocoon. Respiration rates per unit mass of wings of A. pernyi are similar to those of the whole, resting adult. The possible adaptive significance of this tracheation and its variation is discussed.     

Autophagy and apoptosis coordinate physiological cell death in larval salivary glands of Apis mellifera (Hymenoptera: Apidae)

Larval salivary glands of bees provide a good model for the study of hormone-induced programmed cell death in Hymenoptera because they have a well-defined secretory cycle with a peak of secretory activity phase, prior to cocoon spinning, and a degenerative phase, after the cocoon spinning. Our findings demonstrate that there is a relationship between apoptosis and autophagy during physiological cell death in these larval salivary glands, that adds evidence to the hypothesis of overlap in the regulation pathways of both types of programmed cell death. Features of autophagy include cytoplasm vacuolation, acid phosphatase activity, presence of autophagic vacuoles and multi-lamellar structures, as well as a delay in the collapse of many nuclei. Features of apoptosis include bleb formation in the cytoplasm and nuclei, with release of parts of the cytoplasm into the lumen, chromatin compaction, and DNA and nucleolar fragmentation. We propose a model for programmed cell death in larval salivary glands of Apis mellifera where autophagy and apoptosis function cooperatively for a more efficient degeneration of the gland secretory cells.     

Ecdysteroid as a mediator in the regulation of silk protein synthesis and its influence on silkworm (Bom., Lepidoptera) genome

Abstract:  Ecdysteroid is one of the two most important insect hormones, which controls growth and developmental activities of insect. In silkworm, Bombyx mori L., basically ecdysteroid induces molting and metamorphosis in larvae and exogenous administration hastens the silk synthetic activity and cocoon spinning process but it strictly depends on the time of administration. In the present study, we administered phytogenous ecdysteroid to the 48-h-old 5th instar silkworm, B. mori at a single dose of 2  μ g per larva to study its effects on the larval growth, cocoon and silk variables. The possible role of ecdysteroid in altering the developmental sequence of silkworm and thereby its effect on cocoon and silk production at molecular level have been tried to get elucidated. The genomic DNA was isolated from the posterior silk gland on day 5 and 7 of the 5th instar from the ecdysteroid treated and the control larvae and was randomly amplified with arbitrary primers. The result presented notable variation in the amplified product suggesting the participation of ecdysteroid in regulating the silk gene. The feeding period of treated larvae was unaffected while the cocoon characters exhibited considerable improvement. The filament traits also were improved notably in the treated larvae. The participation of ecdysteroid in the silk biosynthetic process with its physiological and molecular implications was discussed.     

Infochemicals influencing the host foraging behaviour of Dahlbominus fuscipennis, a pupal parasitoid of the European spruce sawfly (Gilpinia hercyniae)

The chalcidoid wasp Dahlbominus fuscipennis Zett. (Hymenoptera: Eulophidae) parasitises pupae of the European spruce sawfly Gilpinia hercyniae Htg. (Hymenoptera: Diprionidae). In this parasitoid-host system the role of infochemicals for successful parasitisation was studied. Olfactometer experiments and dual choice bioassays in petri dishes showed that D. fuscipennis was not attracted by volatiles from a plant-host complex consisting of spruce, sawfly larvae and pupae. On the other hand, odours from needles of Picea abies (L.), that were collected from the ground, attracted D. fuscipennis to the habitat of the sawfly. Volatiles from host pupae were not attractive, whereas a contact kairomone from sawfly pupae elicited frequent antennal drumming, a behaviour that indicates host recognition. This contact kairomone was extracted with methanol from the outer cocoon layer of the pupa. The kairomone is not produced by the silk glands of the last instar larva spinning the cocoon, since a methanolic extract of silk gland secretion did not elicit any response in D. fuscipennis. Further bioassays revealed that non-volatile components of the integument of the prepupa within the cocoon also elicited antennal drumming. We suggest that chemicals of the prepupal integument diffuse from inside the cocoon to the outside where they inform the parasitoid on the suitability of the host.     

Effect of temperature and relative humidity on spinning behaviour of silkworm (Bombyx mori.L)

The rate of spinning of silkworm larva (Bombyx mori.L) was slow at 22 degrees C and fast at 38 degrees C. The time taken for completion of cocoon was longest at 98 +/- 2% and least at 40 +/- 2% RH. However, good quality cocoons were spun at 22 degrees C and 65 +/- 5% RH, hence it would be advantageous to maintain this temperature and relative humidity at the time of cocoon spinning in the case of CB race of silkworm.     

THE DEPENDENCE OF CECROPIA YOLK FORMATION IN VITRO ON SPECIFIC BLOOD PROTEINS

The capacity of cecropia vitellogenic follicles to form yolk during short-term in vitro incubation in female blood was analyzed by labeling with fluorescein-conjugated serum globulin, tritiated cecropia blood proteins, or tritiated amino acid. As judged by fluorescence microscopy or autoradiography, yolk formation during 3–8 hr in vitro was similar in rate and in protein uptake specificity to that observed in vivo. When follicles were incubated in cecropia male blood, 6% gamma globulin, or cecropia saline, the yolk produced was markedly inferior in quality and quantity to that generated in female blood. Purified preparations of vitellogenin, the primary female blood protein deposited in the yolk, were equivalent to whole female blood in supporting yolk formation; this protein seems, therefore, to have a specific stimulatory role. An enhancement of the rate of pinocytosis at the oocyte surface by vitellogenin is postulated.