Ascending flights for migration of the oriental armyworm,Pseudaletia separata

Flight behavior ofPseudaletia (= Mythimna)separata Walker (Lepidoptera: Noctuidae) was observed around sunset in a grassland habitat at Morioka in northern Japan. Take-off flights by both females and males commenced on the first day following emergence. The proportion of moths flying increased gradually with age from 62% on day 1 to 86% on day 5. Flight could be categorized as ascending or short range, 2 to less than 10 m flights in the observation. The proportion of adults making ascending flights increased from 47% on day 1 to a maximum of 55% on day 2, and then gradually decreased to 29% on day 5. It is considered that the ascending flight is connected with long-distance migration. Mated females with matured eggs did not make ascending flights, but mated males did. Adults embarked on those flights at temperatures of 7.5°C or greater. On rainy days fewer adults flew upwards than on non-rainy days. The related species,Leucania pallens, did not fly up, but flew at slow speed at low altitude.

---

Résumé Le comportement de vol dePseudaletia (Mythimna) separata Walker (Lep., Noct.) a été observé vers le crépuscule au dessus de prairies à Morioka (Japon septentrional). Les envols des femelles et des mâles débutent le jour suivant l'émergence. La proportion de papillons en vol augmente progressivement, de 62% le premier jour, à 86% le 5e jour. Les vols peuvent être classés en ascendants (2 à 10 m) ou à courte distance (inférieurs à 2 m). La proportion d'adultes effectuant des vols ascendants augmente, de 47% le premier jour, à un maximum de 55% le 2^e jour, pour diminuer progressivement jusqu'à 29% le 5^e jour. On considère que les vols ascendants sont lié aux migrations à longue distance. Les femelles fécondées contenant des ufs mûrs ne font pas de vols ascendants, tandis que les mâles ayant eu une activité sexuelle le font. Les adultes effectuent de tels vols aux températures supérieures ou égales à 7.5°C. Par temps pluveuxmoins d'adultes ont des vols ascendants que par temps sec. L'espèce voisine,Leucania pallens n'effectue pas de vols ascendants, mais vole à faible vitesse à basse altitude.

     

Drosophila hemolymph proteins: Purification,characterization, and genetic mapping of larval serum protein 2 in D. melanogaster

Three of the major protein species present in the hemolymph of Drosophila melanogaster larvae just prior to pupation are absent from second instar larvae but accumulate rapidly during the third instar. This article describes the purification and characterization of one of these, larval serum protein (LSP) 2, using an immunological assay. It is a homohexamer of molecular weight about 450,000, with a polypeptide molecular weight of 78,000–83,000. Fast and slow electrophoretic variants of this protein map between the markers vin and gs, at 36–37 on chromosome 3.This work was partially supported by M.R.C. Research Studentships to J.W. and M.E.A.     

The endocrine-genetic basis of life-history variation: the relationship between the ecdysteroid titer and morph-specific reproduction in the wing-polymorphic cricket Gryllus firmus

The hormonal basis of variation in life-history traits is a poorly studied topic in life-history evolution. An important step in identifying the endocrine-genetic causes of life-history variation is documenting statistical and functional associations between hormone titers and genotypes/phenotypes that vary in life-history traits. To this end, we compared the blood ecdysteroid titer and the mass of the ovaries during the first week of adulthood among a flight-capable morph and two flightless morphs of the wing-polymorphic cricket Gryllus firmus. Ecdysteroids are a group of structurally related hormones that regulate many important aspects of reproduction in insects. Both the ecdysteroid titer and ovarian mass were significantly higher in each of two flightless morphs compared with the flight-capable morph throughout the first week of adulthood. Genetically based differences in the ecdysteroid titer and ovarian mass between morphs from different selected lines were similar to phenotypically based differences among morphs from the same control (unselected) lines. By day 7 of adulthood, ovaries were typically 200-400% larger and the ecdysteroid titer was 60-300% higher in flightless versus the flight-capable morph. In addition, highly significant, positive, phenotypic correlations were observed between the ecdysteroid titer and ovarian mass in pooled samples of the two flightless and flight-capable crickets from control lines or from selected lines. The ecdysteroid titer was sufficiently elevated in the flightless morphs to account for their elevated ovarian growth. This is the first direct documentation that naturally occurring phenotypes/genotypes that differ in early fecundity, a key life-history trait, also differ phenotypically and genetically in the titer of a key reproductive hormone that potentially regulates that trait.     

Ultrastructure of spermiogenesis in Plagioscion squamosissimus (Teleostei, Perciformes, Sciaenidae)

Spermiogenesis in Plagioscion squamosissimus occurs in cysts. It involves a gradual differentiation process of spermatids that is characterized mainly by chromatin compaction in the nucleus and formation of the flagellum, resulting in the spermatozoa, the smallest germ cells. At the end of spermiogenesis, the cysts open and release the newly formed spermatozoa into the lumen of the seminiferous tubules. The spermatozoa do not have an acrosome and are divided into head, midpiece, and tail or flagellum. The spermatozoa of P. squamosissimus are of perciform type with the flagellum parallel to the nucleus and the centrioles located outside the nuclear notch.     

蒲氏钩蝠蛾幼虫血淋巴物质与其耐寒性

作为冬虫夏草寄主,蒲氏钩蝠蛾具重要的经济研究价值,主要分布于青藏高原色季拉山,具有较强的耐寒性,其耐寒性机理尚不明了。通过调查蒲氏钩蝠蛾生境土壤温度与血淋巴物质组成周年变化,以期探讨其耐寒性机理。实验结果表明:土壤温度呈规律性升降趋势,1、2月最低,8月逐步上升至最高,而后逐渐下降;血淋巴中主要物质按含量从高到低排列依次为:蛋白质(8.37—74.2 mg/m L)、总糖(12.65—36.12 mg/m L)、脂肪(7.7—12.32 mg/m L)以及甘油(1.66—3.91 mg/m L);在温度较高的夏季,蛋白质、总糖以及脂肪等物质含量处于较低水平,相反,在寒冷的冬季,此三者的含量均居较高水平;此3类能源物质含量与野外土壤温度均呈极显著的负相关关系,因而血淋巴总热量值在夏季较低而冬季较高,与野外土壤温度亦呈极显著负相关关系;与之相反的是,血淋巴中甘油含量的周年变化规律与野外土壤温度无明显的相关关系。因此,血淋巴中蛋白质、总糖以及脂肪等物质对于蒲氏钩蝠蛾耐寒性有着非常重要的作用,幼虫通过积累3类物质,增加血淋巴能量值,抵御青藏高原的高寒气候;可能由于生境的不同,甘油在蒲氏钩蝠蛾中的作用与其它昆虫有差异,在耐寒性提高中的功能尚需进一步研究。实验结果为了解蒲氏钩蝠蛾的强耐寒性以及适应高寒环境的生理生化机制提供理论依据。     

Genetics of the hemolymph esterases ofLucilia cuprina (Diptera: Calliphoridae)

When hemolymph from adults ofLucilia cuprina was partitioned on native polyacrylamide gels, nonspecific esterase staining demonstrated 10 bands with up to six bands in an individual. The bands derive from alleles at two loci,E _HA (five alleles) andE _HB (four alleles).E _HA is located on chromosome 4, 16.3 map units fromsv (singed vibrissae) and 22.1 map units fromra (radial vein gaps).E _HB is located on chromosome 5, 34.0 map units fromto ² (topaz² eyes) and 7.2 map units frommv (M1-veinless).     

Host frass as arrestant chemicals in locating host Mythimna separata by the tachinid fly Exorista japonica

We investigated the response of the tachinid fly, Exorista japonica (Townsend), to host frass or its extracts in order to clarify the host location mechanisms of female flies in a potential host habitat. Host searching time in a patch and the number of patch visits were analyzed by using a frass-containing patch which was excreted by host larvae, Mythimna separata (Walker) (Lepidoptera: Noctuidae), and patches to which host frass extracts were applied. E. japonica females were arrested in response to the host-frass-containing patch after contacting the frass with their front tarsi, thereby spending most of the time to search the patch and to revisit the host-frass patch. While host-searching time in the patch by the females was longest at their first visit of a patch with host frass, searching time decreased with successive visits. The female flies also exhibited area-restricted searching with methanol extracts of the host frass. Area-restricted searching activity increased with the concentration of host-frass extract, i.e., total searching time in the patch and the number of patch visits varied in a dose-dependent manner. E. japonica females likely employ chemicals in host frass as arrestants in host location.     

Site of synthesis and phylogenetic distribution of a hemolymph trophic factor of the tobacco hornworm,Manduca sexta

Summary Identification of fith instar larvalManduca sexta fat body and epidermis as sites of synthesis of a hemolymph protein (hemolymph trophic factor or HTF) was achieved using in vitro³H-leucine incorporation into protein and subsequent immunoprecipitation of tissue homogenates. Fat body is the primary site of HTF synthesis with a maximal rate on Day 1; epidermis is a secondary site with peak synthesis on Day 0. In vitro radiolabelling followed by TCA precipitation of general protein of fat body and epidermal homogenates suggest that fat body actively elaborates protein on Days 0–5 with peak rates on Days 1 and 4, while epidermis is active on Days 0–5 with a peak rate on Day 3. Based on Anti-HTF ELISA estimates, HTF [500 to 1000 μg/ml] was found in the hemolymph of representatives of the insect orders Blattodea, Hemiptera, Orthoptera, and Lepidoptera and in the class Crustacea, but not in the class Merostomata. These studies suggest a possible fundamental role for HTF among modern arthropods in cuticular deposition involving both epidermis and fat body. The physiological role of HTF is undetermined.     

Acid phosphatase activity in hemolymph of the migratory grasshopper,Melanoplus sanguinipes, duringBeauveria bassiana infection

The distribution and abundance of acid phosphatase (AP) in hemolymph (HL), plasma (PM) and hemocyte lysate supernatant (HLS) of the migratory grasshopper,Melanoplus sanguinipes infected withBeauveria bassiana (strain GK 2016) has been examined. AP activity was determined at intervals from 30 min to 60 h postinjection of 2 μl of 1×10⁸ conidia/ml per grasshopper. The enzyme was detected with the substrate β-glycerophosphate in sodium acetate acetic acid buffer form hemocytes (HC) and withp-nitrophenol phosphate sodium salt for HL, PM and HLS. In results of experiment 1 proportion of HC showing AP activity increased 1–2 h, then returned to normal after 4 h. However, inB. bassiana-injected grasshoppers, a second increase was noted 24 h later which was not seen in the Tween-80-injected insects. Uninjected controls showed no change with time in the proportion of HC with AP activity. Studies were also made of the distribution of AP activity in the HL, PM, and HLS. AP activity in HL appeared to vary with the sex of the grasshoppers. Females showed increase in AP activity in HL 18–24 h after injection withB. bassiana, whereas males only showed an increase 1 h after injection. Assay of HLS showed that the level of AP activity did not change significantly throughout the experiment. Changes in AP activity in PM, in bothB. bassiana — and Tween-80-injected insects (both sexes) paralleled those of the HL, indicating that the enzyme is released from the HC. The observations are discussed in terms of the possible role of AP in the immune response ofM. sanguinipes.     

Developmental changes in hemolymph ecdysteroid level and prothoracicotropic hormone activity during the fifth larval instar of the Eri silkworm, Samia cynthia ricini

Developmental changes in hemolymph ecdysteroid level, ecdysteroid synthesis by prothoracic glands （PGs） in vitro, prothoracicotropic hormone （PTTH） activity in brain extracts, and PTTH activity in the hemolymph were measured during the fifth larval instar of the Eri silkworm, Samia cynthia ricini. The changing patterns of hemolymph ecdysteroid level and ecdysteroid synthesis by laGs in vitro are similar to each other, with maximums on day 9. However, on this day, hemolymph ecdysteroid level was substantially higher than ecdysteroid synthesis by PGs in vitro suggesting a high PTTH activity in the hemolymph on day 9. Moreover, the changing pattern of PTTH activity in brain extracts is also similar to that of PTTH activity in the hemolymph, both peaking on day 9. However, on this day, activity in brain extracts was much smaller than PTTH activity in the hemolymph implying that most PTTH synthesized by the brain is secreted to the hemolymph and the brain stores a very little amount of PTTH. This study provides unique insights onto the hormonal regulation of ecdysteroid synthesis in the Eri silkworm and is useful for our future studies on signal transduction of insect neurolaelatides.     

Caste-specific modulation of juvenile hormone III content and ecdysteroid titer in postembryonic development of the stingless bee,Scaptotrigona postica depilis

Summary Juvenile hormone III content and ecdysteroid titer were analyzed for larval and pupal development of the stingless bee,Scaptotrigona postica depilis. Castespecific differences in juvenile hormone III content were detected at three developmental phases: at the transition from the fourth to the fifth larval stadium, in the spinning phase of the fifth larval stadium, and shortly after the imaginal moult. During the fifth larval stadium, juvenile hormone content closely reflects corpora allata activity. Juvenile hormone synthesis may thus be responsible for the elevated hormone titer in spinning-phase queen larvae, a phase of known sensitivity for induction of queen characters by exogenous juvenile hormone. For ecdysteroids, two phases of caste-specific differences were found: in the pre-pupal phase, and shortly after the imaginal moult. In both periods the titer in queens is distinctly higher compared to workers.Abbreviations Im imago 1 day after eclosion - L3, L4, L5 larval instars 3, 4, and 5 - L5F1, L5F2 substages of feeding phase in fifth larval instar - L5S1, L5S2, L5S3 substages of spinning phase in fifth larval instar - PP1, PP2 substages of prepupal phase - Pw white eyed pupa - Pp pink eyed pupa - Pr red eyed pupa - Pd dark eyed pupa - Pdl, Pdm, Pdd dark eyed pupa with progressive tanning of cuticle - RIA radioimmunoassay     

Purification and properties of the melanization and reddish colouration hormone (MRCH) in the armyworm,Leucania separata (Lepidoptera)

Larvae of Leucania separata show a marked black colouration under crowded conditions due to the action of the neurohormone termed melanization and reddish colouration hormone (MRCH). MRCH was extracted with 80% ethanol from larval brains (Br) or suboesophageal ganglia (SG) of L. separata and Br-SG of the silkworm moth, Bombyx mori. Approximately a 16,300-fold purification of the extract derived from 192,000 Bombyx heads afforded two active fractions, each of which caused a remarkable melanization of the cuticle of an L. separata larva. The MRCH activity contained in each fraction is due to a peptide (mol. wt. 6400–8000). Partially purified MRCH also caused melanization in Leucania loreyi, Spodoptera litura, and Mamestra brassicae, but had no diapause hormone activity in B. mori.     

Location of ecdysteroid 22-O-acyltransferase in the larvae ofHeliothis virescens

Larvae of the tobacco budworm,Heliothis virescens, are resistant to high levels of ingested 20-hydroxyecdysone which could cause potential inhibition to the development of many other lepidopteran species. This resistance is attributed to the ability of the larvae to metabolize this molting hormone to its 22-acyl ester forms. When tobacco budworm larvae were fed large quantities of 20-hydroxyecdyone, the hormonal metabolites were found in gut and fat body tissues. When incubated with 20-hydroxyecdysone gut tissue converted 20-hydroxyecdysone into its 22-acyl ester metabolites. Lumen site of the midgut was found to be the major location of this bio-transformation. In contrast, fat body tissue failed to convert 20-hydroxyecdysone to 22-acyl ester metabolitesin vitro. After the oral injection of³H-ecdysone, the major metabolites formed were ecdysone 22-acyl esters whereas the majority of³H-ecdysone was transformed to polar metabolites after it was injected into the hemocoel of the larvae. Similar distributions of ecdysteroid 22-O-acyltransferase and alkaline phosphatase activity in subcellular fractions demonstrates the co-localization of these enzymes in plasma membrane of the gut epithelial cells. These results suggest that gut brush border membrane is the major site of ecdysteroid 22-acyl ester formation inH. virescens larvae.     

Starvation influences allatotropin gene expression and juvenile hormone titer in the female adult oriental armyworm, Mythimna separata

The first day of adult life is the sensitive stage for shifting migrants into the resident morphs of the oriental armyworm (OAW), Mythimna separata (Walker). The juvenile hormone (JH) titer, expression of the allatotropin (AT) gene, and their relationship were investigated in adult female migrants starved in the sensitive stage, to understand the underlying mechanism of changing migrants into resident OAWs. Haemolymph JH titers of the starved female adults were mostly elevated earlier than in controls, although not all differences were statistically significant. JH I titers in the starved moths were significantly higher than those in the controls on 1, 2, and 5 days after treatment (DAT), respectively. JH II titers in the starved moths were significantly higher than the controls through the period tested except on 5 DAT. JH II is the most likely regulator in changing migrants into resident morphs. The relative quantities of AT expression in the starved moths were higher through the period tested except on 5 DAT. AT expression and JH titers appear to be positively correlated, especially for those in earlier days of the adult life. We infer that AT is the important regulator of JH levels. A model for the shifting of migrants into resident morphs in the OAW is proposed.     

The relationship between outbreaks, rainfall and low density populations of the African armyworm, Spodoptera exempta, in Kenya

This study has demonstrated that low numbers of African armyworms (Spodoptera exempta) (Walker) (Lepidoptera: Noctuidae) can be found throughout the year in parts of the coastal and highland regions of Kenya where there is frequent rainfall. During the study period the numbers of moths caught in pheromone traps in these regions built up during the short and long rains, and decreased dramatically during the intervening dry seasons. There was a lag of one to two months between the peaks for rainfall and moth numbers. This contrasts with the situation at sites of the seasonal outbreaks of armyworm larvae, where a sudden preceding influx of moths coincides with the rainfall. A positive correlation was found, for the long rains seasons only, between the number of armyworm outbreaks throughout the country and the peak numbers of moths in coastal and highland regions.It is suggested therefore that the peak numbers of moths trapped in the eastern highlands and coastal regions during the rainy seasons arise principally from outbreaks in the extensive seasonal grasslands. These grassland areas diminish considerably during the prolonged dry periods between the two rainy seasons, and it is only the eastern highland and coastal regions that are likely to provide suitable habitats for breeding during the dry seasons. The significance of these populations for initiating the first outbreaks of the following season is discussed.     

Aplysia hemolymph promotes neurite outgrowth and synaptogenesis of identifiedHelix neurons in cell culture

Hemolymph of adultAplysia californica significantly affects neurite outgrowth of identified neurons of the land snailHelix pomatia. The metacerebral giant cell (MGC) and the motoneuron C3 from the cerebral ganglion and the neuron B2 from the buccal ganglion ofH. pomatia were isolated by enzymatic and mechanical dissociation and plated onto poly-l-lysine-coated dishes either containing culture medium conditioned byHelix ganglia, or pre-treated withAplysia hemolymph. To determine the extent of neuronal growth we measured the neurite elongation and the neuritic field of cultured neurons at different time points.Aplysia hemolymph enhances the extent and rate of linear outgrowth and the branching domain ofHelix neurons. With the hemolymph treatment the MGC neuron more consistently forms specific chemical synapses with its follower cell B2, and these connections are more effective than those established in the presence of the conditioned medium.     

Association of a fungal endophyte in perennial ryegrass with antibiosis to larvae of the southern armyworm, Spodoptera eridania

Larvae of the Southern armyworm Spodoptera eridania Cramer (Lepidoptera:Noctuidae), feeding on perennial ryegrasses (Lolium perenne L.) infected with an endophytic fungus (Acremonium loliae Latch, Christensen and Samuels), had a much lower survival rate (7–13%) than larvae feeding on endophyte-free ryegrasses (82–90%). Death of the larvae on endophyte-infected entries occurred rapidly between 144 h and 168 h of feeding. This corresponded with armyworms feeding on the base of the plant, where endophyte concentration is highest. Twenty-four hours after the start of the bioassay the larval mass and rate of larval development were significantly higher on endophytic entries. From 48–144 h no differences were seen, but after 144 h the mass of larvae on endophyte-infected grasses sharply declined. Observations from this bioassay further substantiate the association between A. loliae-infected ryegrass and antibiosis to several lepidopterous and coleopterous insect pests.

---

Résumé Le ray-grass vivace (Lolium perenne), contaminé par le champignon endophyte Acremonium loliae Latch, Christensen & Samuels, a présenté une augmentation de la resistance à de nombreux coléoptères et lépidoptères nuisibles. Cette note examine les réactions de Spodoptera eridania Cramer (Lépido., Noctuidae) alimenté sur trois lignées de ray-grass contaminées par le champignon et trois lignées saines. Après 168 h d'alimentation sur ray-grass contaminé, les chenilles presentent une très forte mortalité; la survie n'est que de 7 à 13% contre 82 à 90% pour le ray-grass sain. Le décès brutal des chenilles correspond à leur alimentation sur la base de la plante ou la concentration du champignon est la plus forte. Les chenilles consomment constamment, broutant plus des 2/3 du feuillage du ray-grass; les broutements des six séries ne différaient pas significativement.Au bout de 24 h, la nombre de chenilles passées du 3ème au 4ème stade, et l'augmentation de poids sont plus élevés pour les séries sur plante contaminée, ce qui suggère un avantage initial pour les chenilles en présence de champignon endophyte, l'analyse en poids sec a montré que l'augmentation de poids initial est réel. Entre 48 et 144 h, cependant, le nombre de 4ème stade et le poids des chenilles sont les mêmes dans les deux séries. Après 144 h, le poids des chenilles sur ray-grass contaminé diminue significativement; aucune n'était parvenue au 5ème stade, contre 11% sur ray-grass sain. Nous n'avons pas observé de signes apparents de neurotoxicité. Au lieu de cela, il ya a eu interaction avec un processus physiologique fondamental, ce qui a provoqué une forte perte de poids larvaire et la mort, indiquant l'intervention d'inhibiteurs métaboliques.