Factors controlling in vitro hatching of Vairimorpha plodiae (Microspora) spores and their infectivity to Plodia interpunctella,Heliothis virescens, and Pieris brassicae

Factors which influence the hatching of spores and proliferation of stages of the microsporidium Vairimorpha plodiae in two susceptible insects, Plodia interpunctella and Heliothis virescens, and one nonsusceptible insect, Pieris brassicae, were investigated. Spores hatched in 0.1 and 1 m KCl solutions when subjected to a change in pH, from pH 11 to pH 8. K⁺ was essential for hatching; NaCl solutions were not effective. Ca²⁺ and Mg²⁺ inhibited hatching, and calcium and magnesium chelating agents enhanced it. All three insect species had alkaline midgut contents and smooth, fragile peritrophic membranes. Spores hatched inside the midguts of all three insect species (P. interpunctella: maximum rate, 92.5%; H. virescens, 91.5%; P. brassicae, 82%). Sporoplasms were observed in the midgut epithelial and associated tracheole cells of P. brassicae. Both H. virescens and P. brassicae became infected when injected intrahemocoelically with spores.     

A redescription of Nosema herpobdellae (Microspora: Nosematidae), a parasite of the leech Erpobdella octoculata (Hirudinea: Erpobdellidae)

Nosema herpobdellae was recorded in populations of the leech Erpobdella octoculata from lakes in northwest England and North Wales and is redescribed using light and transmission electron microscopy. It differes from N. glossiphoniae in the nature of the infection and tissues parasitized and from N. tractabile in its larger spore size, longer polar filament, in the angle of the anterior coils of the polar filament to the spore long axis, and apparently in its developmental cycle. The infection was found in a massive xenoma, in the connective tissue surrounding the gut, which was presumed to be formed from a single hypertrophied cell. Its developmental cycle included merogony and sporogony.     

Importance of timing,spore concentrations,and levels of spore carrier in applications of Nosema locustae (Microsporida: Nosematidae) for control of grasshoppers

In a study of microbial control of grasshoppers, Nosema locustae was applied to 10-acre plots in 2 replications of 24 treatment combinations in which the factors were (1) times of applications: 4 applications spaced over 22 days; (2) concentrations of spores: 1, 30, and 900 spores/in²; and (3) levels of the spore carrier: 1 and 4 lb wheat bran/acre. The concentrations of spores had the most effect in reducing the densities of grasshoppers and the incidence of infection among the survivors at the last sampling (coincided with the initiation of oviposition), the average reduction in density attributed to application of 1 spore/in² was 21%, that attributed to 30 spores/in² was 46%, and that attributed to 900 spores/in² was 73%. The first and second applications caused the greatest reductions in density; the second, third, and fourth applications produced the highest incidence of infections. Also, a higher incidence of infection was observed among grasshoppers from plots treated with 30 spores/in² on 1 lb bran/acre than from plots treated with 30 spores/in² on 4 lb of bran/acre. Therefore, a ratio of about 0.63–0.94 billion spores/lb bran applied at a rate of 1–1.5 lb/acre (= 100–150 spores/in²) at the time when the principal early summer species are third-instar nymphs would have resulted in overall reductions in density of 50–60%, with 35–50% of the survivors sufficiently infected so that fecundity would probably be affected. The reductions and incidence of infections would be higher in species that readily accept and use wheat bran, among which are some of the more economically important species of grasshoppers.     

Single cell responses in female Pieris brassicae (Lepidoptera: Pieridae) to plant volatiles and conspecific egg odours

Action potentials from individual olfactory cells and receptor potentials were recorded from antennae of female cabbage white butterflies, Pieris brassicae, using a new ‘surface-contact’ recording technique. Stimuli used were plant volatiles and conspecific egg odours. Most cells were activated by some stimuli and inhibited by others. Hence, the ‘odour spectra’ of most cells included activating as well as inhibiting substances. In addition, the ‘response profile’ of a given stimulus contained both positively and negatively responding cells.Cluster analysis of the odour spectra revealed the existence of moderately separated clusters of olfactory receptors. No groups of cells specifically tuned to odours of eggs or cabbage leaves were found. Analysis of the response profiles did not show a clear clustering of stimuli. The results suggested that the membranes of different receptor cells contain several types of acceptor sites in various proportions.No correlation between EAG and receptor potential amplitude or spike frequency was found. It was concluded that the EAG may be useful in comparing the relative receptor sensitivities only when chemically related compounds are involved.     

Infectivity of a microsporidium of mosquitoes (Nosema algerae) to larval stages of Schistosoma mansoni in Biomphalaria glabrata

Lai P. F. and Canning E. U. 1980. Infectivity of a microsporidium of mosquitoes (Nosema algerae) to larval stages of Schistosoma mansoni in Biomphalaria glabrata. International Journal for Parasitology10: 293–301. Nosema algerae derived from a closed colony of Anopheles stephensi was fed to Biomphalaria glabrata infected with Schistosoma mansoni. Mother and daughter sporocysts became hyperinfected but the snail tissues remained free of the microsporidia except for rare small aggregates of spores. These lay close to the sites occupied by mother or daughter sporocysts and were probably liberated from them. Irrespective of dose, fewer snails contained infected sporocysts when spores were given at 7 days post-miracidial infection than when given at 14 days. These periods corresponded respectively to stages when mother sporocysts only or daughter sporocysts as well were present in the snails. Infection of the sporocysts began in the tegumental cells, spread to the brood chamber and ultimately to the cercariae themselves. Heavily infected sporocysts contained fewer developing embryos. Doses of 10⁶ and 10⁷ spores/snail caused significant depression of cercaria output when given at 14 days but not at 7 days.     

The effects of actinomycin D,cycloheximide and puromycin on the 20-hydroxyecdysone induced autophagocytosis in larval fat body cells of Pieris brassicae

The effects of 20-hydroxyecdysone (5 μg/g body weight), cycloheximide (5 μg/g body weight), puromycin (30 μg/g body weight) and actinomycin D (30 μg/g body weight) were investigated on the larval fat body cells of 48–70-hours-old last instar larvae of Pieris brassicae.20-Hydroxyecdysone was found to induce the formation of autophagic vacuoles 3 hr after its administration, but this was, however, prevented by simultaneous cycloheximide treatment in a parallel experiment. On the contrary, puromycin proved to induce autophagocytosis. These diverse effects of the two translational inhibitors on hormone-induced autophagocytosis may be explained by differences in their modes of action.Actinomycin D, when administered 21 hr prior to the hormone, exerted a preventive effect on induced autophagocytosis, but was ineffective when applied 3 hr before the injection of 20-hydroxyecdysone.It was concluded that the presence of RNA synthesized several hours prior to the hormone treatment was a prerequisite for induction of autophagocytosis by 20-hydroxyecdysone.     

Haemolymph protein reserves of diapausing and nondiapausing codling moth larvae, Cydia pomonella (L) (Lepidoptera: Tortricidae)

Haemolymph from diapausing codling moth larvae contain a dominant soluble protein, which is of minor significance in the haemolymph of nondiapausing larvae. This haemolymph protein is accumulated during the ultimate larval instar and its concentration is maintained throughout diapause development. Larvae induced into a diapause-like condition following topical treatment with a juvenile hormone mimie stored the haemolymph protein.     

The replication of a cytoplasmic polyhedrosis virus from Chrysodeixis eriosoma (Lepidoptera: Noctuidae) in Spodoptera frugiperda cells

A cytoplasmic polyhedrosis virus (CPV) from Chrysodeixis eriosoma (Lepidoptera: Noctuidae) replicated in Spodoptera frugiperda cells. Low rates of infection were achieved, even at high multiplicities of infection and TCID₅₀ assays showed that there was negligible release of virus particles from infected cells. In an infected focus assay, based on formation of PIB, the dose-response data demonstrated that a single particle could initiate infection. No loss of infectivity occurred in virus preparations stored at 4°, ?20°, or ?90°C, but infectivity of virus stored at 20°C declined sharply. A small isometric virus contaminant was present in some CPV preparations and its interaction with the CPV is discussed. Limited CPV infection was achieved in Trichoplusia ni cells, but attempts to infect Aedes aegypti cells were unsuccessful.     

Life cycle,epizootiology, and horizontal transmission of Amblyospora (Microspora: Amblyosporidae) in a univoltine mosquito, Aedes stimulans

Amblyospora infections in Aedes stimulans are transovarially transmitted by females infected in the previous year. Pathogen development in progeny is dimorphic and host sex dependent. In males, the pathogen invades fat body tissue and undergoes an extensive developmental sequence which kills the host and results in the formation of eight haploid spores enclosed in an accessory membrane that are not infectious to other larvae. In females, the pathogen invades host oenocytes and undergoes a simple developmental sequence which has no detrimental affect on longevity, fecundity, oviposition, or egg hatch, and results in the formation of binucleated spores that infect the ovaries and ensure transmission to the next generation. Transovarial transmission is continuous and is the major way in which these microsporidia are maintained from year to year, but is incapable of maintaining infections in breeding populations because of low transmission rates and is not sufficient to account for the types and levels of infection observed in the field. Horizontal transmission is reported for the first time. It occurs sporadically during the early stages of larval subsequently disseminated to oenocytes of adult hosts and are transovarially transmitted by females to filial host generations. This pathway of transmission provides the necessary mechanism whereby these microsporidia can reenter the mosquito population and thus perpetuate themselves.     

Incubation period, spore egestion and horizontal transmission of Nosema fumiferanae (Microsporidia: Nosematidae) in spruce budworm (Choristoneura sp., Lepidoptera: Tortricidae): the role of temperature and dose

Various instars of Choristoneura occidentalis were fed with a range of doses of Nosema fumiferanae and reared at 20, 24 and 28 degrees C to determine the influence of temperature and dose on the time to spore egestion and the number of spores egested in the frass. When larvae were fed in the third stadium, as few as 10(2) spores per larva initiated infection, and both onset of spore egestion and the number of spores egested were affected by a complex relationship between temperature and inoculation dose. Onset of spore egestion varied from 11 to 15 days postinoculation. At 20 degrees C, the onset was delayed and spore production decreased with increasing inoculation dose whereas at higher temperatures spores were first egested at the lowest dose and spore production increased with dose. When larvae were fed spores in the fifth and sixth stadium, no spores were egested because pupation occurred before completion of the incubation period. To assess the effect of temperature on horizontal transmission, Choristoneura fumiferana larvae fed with 10(4) N. fumiferanae spores per larva were reared with uninfected larvae at 15, 20 and 25 degrees C. At 15 degrees C, we observed the highest degree of horizontal transmission, defined by the largest change in N. fumiferanae prevalence, even though the density of spores available for horizontal transmission was the lowest. Infected adults eclosed later than uninfected adults and the time to eclosion was also dependent on sex and temperature. We relate our experimental findings to consequences for horizontal and vertical transmission of N. fumiferanae in spruce budworm populations.     

Environmental factors affecting black/white coloration of the silken girdle in the swallowtail butterfly, Atrophaneura alcinous (Lepidoptera: Papilionidae)

The silken girdles of pupae of the swallowtail butterfly Atrophaneura alcinous show black and white color diphenism. Field observations revealed that all pupae observed on non-food plants and the leaves and stems of the larval food plant Aristolochia debilis were classified as a silken girdle of a black type, while a large portion of pupae pupating on the twigs and trunks of cherry trees in close proximity to A. debilis were classified as a silken girdle of a black type. Additionally, all pupae observed on the surfaces of artificial objects in areas where there are no surrounding plants or trees were classified as a silken girdle of a white type. We demonstrated the effect of day length and the texture, light, plant odor and humidity of pupation sites on the coloration of the silken girdle in A. alcinous. Regardless of long-day or short-day day length conditions, light conditions of constant light or dark, or the presence of a plant odor of A. debilis as environmental cues, all larvae placed at over 80% relative humidity (R.H.) developed into pupae with a silken girdle of a black type. However, all larvae developed into pupae with a silken girdle of a white type when R.H. was below 75%. Furthermore, when pupae with a silken girdle of a white type were transferred to conditions of 90% R.H. within 24 hr of pupation, the white color of the silken girdle changed into a black type within 24 hr of the transfer. The present data suggest that the induction of a black coloration of the silken girdle in A. alcinous requires a R.H. of approximately 80% or more as an environmental factor.     

Some factors affecting flight in field populations of the Australian plague locust, Chortoicetes terminifera (walker), in New South Wales

An attempt was made to determine some of the factors regulating flight activity in the Australian plague locust, Chortoicetes terminifera (Walk.) by comparing the physiological characteristics of flying and non-flying locusts at different times of the day. Flight during the day did not appear to involve extensive displacements of the insects and was dependent largely on the, environmental conditions. The physiological state and age did not seem important and the milling flight of swarms involved insects of all ages and states of sexual maturity and the insects were commonly fully fed. There was probably at this time a continual interchange of insects between the air and the ground provided wind speeds are not too high and the air temperature was not below 19°C. Night flight, however, was observed only in locusts after the teneral period, when cuticle deposition was nearing completion around 10 days after the imaginal ecdysis and, in the case of the females, the oocytes were relatively undeveloped. Night flight was also associated with a failure to feed prior to sunset; only those with empty foreguts being stimulated to fly with the drop in light intensity, when the temperature was not too low and certainly not below 17·5°C. It is probably the older insects which take-off in these circumstances for by the time of sunset 30 to 40 per cent of the older, ones apparently fail to feed on any one evening and empty locusts tend to be more active than fully fed ones. The females appeared to show a flight periodicity related to the oviposition cycle, which under suitable conditions, leads to the displacement of about one-third of the population which is sexually immature or which has already recently oviposited. The flight activity of females is probably greater than that of the males for the sexes were in equal ratio in insects caught in flight while males exceeded females in ground populations. At sunset, fewer females than males still had full foreguts and were perhaps more likely to take-off while a higher proportion of younger females than males may soon re-alight if their physiological state is unfavourable for prolonged flight at night.     

Acylamines enhance the infection of a baculovirus of the armyworm, Pseudaletia unipuncta (Noctuidae,Lepidoptera)

The acylamines (cationic detergents) enhance the infection of a nuclear polyhedrosis virus (NPV) in larvae of the armyworm, Pseudaletia unipuncta. Those with 6 carbons (hexylamine) or 12 carbons (dodecylamine) in the acyl group are more effective in enhancing the NPV than those with 4, 8, or 10 carbons. Hexylamine and dodecylamine increase the rate of infection beyond the 50% level (ID₅₀) by 10 to 100 times when based on the concentration of the virus polyhedra (inclusion bodies).     

Gonadectomy,sex hormones and the growth of tetrathyridial populations of Mesocestoides corti (Cestoda: Cyclophyllidea) in mice

Gonadectomy in mice considerably depressed the volumes of intraperitoneal tetrathyridial populations, orchiectomy being more efficient than ovariectomy. Testosterone propionate accelerated the growth of tetrathyridial populations in gonadectomized mice of both sexes. Oestradiol benzoate was less efficient, but considerably increased the invasion of livers both in intact and in gonadectomized mice. The average size of tetrathyridia was inversely proportional to the size of the populations: tetrathyridia from an orchiectomized mouse were the largest, those from an orchiectomized and testosterone treated mouse, the smallest, and those from an orchiectomized mouse treated with oestradiol, intermediate in size.     

Effect of several factors on the phototactic response of the oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae)

Recently, light traps equipped with light-emitting diode (LED) lights have been used to monitor and control nocturnal insect pests. The oriental armyworm, Mythimna separata Walker, is an important cosmopolitan agriculture pest. The phototactic responses of nocturnal insects, including moths are influenced by abiotic and biological factors. The present study aimed to evaluate the phototactic response to different wavelength LED lights to determine the wavelength to which M. separata adults are most sensitive, and then identify the effects of several factors on the phototactic response. Our experiments revealed that M. separata adult moths had a significantly higher attraction rate to the green (520?nm) LED light compared with other LED lights, the green LED light was 1.28 times more attractive the moths than the commercial double-wave light used as a control. The phototactic response of the adult moths was significantly influenced by several factors, including adaption time in darkness, age, mating status and sexuality. These findings suggest that using a green LED light as a light source in trap could be effective in monitoring or controlling M. separata moths. Our results may provide a theoretical and scientific basis for improving the light trap technique for M. separata moths.     

Repeated larval diapause and diapause-free development in geographic strains of the burnet moth Zygaena trifolii Esp. (Insecta,Lepidoptera)

Summary Zygaena trifolii is a long-day insect with temperature-dependent photoperiodic responses. All larval instars are sensitive to photoperiod; however, diapause may occur at the third larval stage or any subsequent larval instars. There were quantitative differences within populations in the threshold photoperiod for diapause induction. The diapause response was polymorphic, so that larvae might enter diapause at different instars under the same culture conditions. Furthermore, decreasing photoperiods below a critical daylength shifted the diapausing instar towards earlier stages. Geographic strains of Z. trifolii showed discontinuous clinal variation. Near the northern edge of the distribution [Cologne (Köln), FRG], there is first an obligatory diapause, mainly during early instars, and additional facultative (repeat) diapauses during later larval instars in subsequent years. In the southern part of its distribution, this burnet moth is partially bivoltine in the field with a facultative first developmental arrest and a decreased capacity for repeated diapause (Valencia, Spain; Marseille, France). Further experiments indicated that the photoperiodically controlled diapause reaction is also influenced by the number of photoperiodic cycles experienced during the period spent in each larval instar, which depends on temperature. The adaptive significance of obligatory and facultative repeated diapause, varying even among the offspring of a single female, may be to buffer the populations against the more extreme and, from year to year, unpredictable fluctuations in climatic conditions at the northern edge of the distribution.Abbreviations L₃ feeding 3rd larval instar - L₄D diapausing 4th larval instar - L₅D2 repeat-diapausing larval instar with second diapause at the 5th larval stage - LD light-dark cycle - KT shortday conditions (e.g. LD 8:16) - LT long-day conditions (e.g. LD 16:8)     

Influences of daylength and temperature on the period of diapause and its ending process in dormant larvae of burnet moths (Lepidoptera,Zygaenidae)

The onset of larval diapause in the burnet moth Zygaena trifolii is clearly characterized by the larva molting into a specialized dormant morph. In a potentially bivoltine Mediterranean population (Marseille) two types of diapause can occur within 1 year: firstly, a facultative summer diapause of 3–10 weeks, and secondly, an obligate winter diapause, which can be lengthened by a period of thermal quiescence to several months in temperatures of 5°C. For the first time, three successive physiological periods have been experimentally distinguished within an insect dormancy (between onset of diapause and molting to the next non-diapause stage), using chilling periods of 30–180 days at 5°C, and varying conditions of photoperiod and temperature. These stages are: (1) a continuous Diapause-ending process (DEP); (2) thermal quiescence (Q); and finally, (3) a period of postdiapause development (PDD) before molting to the next larval instar. The result of transferring dormant larvae from chilling at 5°C to 20°C depended on the length of the chilling period. After chilling for 120–180 days, molting to the next instar occurred after 6–10 days, independent of daylength. This period corresponds with the duration of PDD. After shorter chilling periods (90, 60, 30 days and the control, 0 days) the period to eclosion increased exponentially, and included both the latter part of the previous diapause process and the 6–10 day period of PDD. However, photoperiod also influences the time to eclosion after chilling. Short daylength (8 h light / 16 h dark: LD 8/16) lengthened the diapause in comparison to long daylength (16 h light / 8 h dark: LD 16/8). Short daylength had a similar effect during chilling at 5°C, as measured by the longer time to eclosion after transfer. The shorter time to eclosion resulting from longer chilling periods (30–90 days) demonstrates that the state of diapause is continuously shortened at 5°C, and corresponds to the neuroendocrine controlled DEP. Presumably the DEP has already started after the onset of diapause. When chilling was continued after the end of the DEP, which ranged between 90 and 120 days, thermal quiescence (Q) followed (observed maximum 395 days). Different photoperiodic conditions during the pre-diapause inductive period modified diapause intensity (measured as the duration of diapause), in that a photoperiodic signal just below the critical photoperiod for diapause induction (LD 15/9) intensified diapause. Experiments simulating the summer diapause showed that PDD occurred in the range of 10–25°C. Higher temperatures (15 and 20°C) shortened the DEP at LD 16/8, so that at 20°C many individuals had already terminated diapause after 10–40 days and had molted after the 6–10 days of PDD. A temperature of 25°C unexpectedly lengthened the DEP to 110 days in several individuals. The ecological consequences and the adaptive significance of variation in the duration of the diapause are discussed in relation to the persistence of local populations predictably variable and rare climatic extremes throughout the year.     

Proteomic analysis of nucleopolyhedrovirus infection resistance in the silkworm, Bombyx mori (Lepidoptera: Bombycidae)

Silkworm hemolymph is an important defense tissue to resist bacteria and virus infections. To study the response of silkworm hemolymph in the resistance of Bombyx mori L. nucleopolyhedrovirus (BmNPV), we constructed a near-isogenic silkworm line with BmNPV resistance using highly resistant and highly susceptible parental strains. In this paper, two-dimensional gel electrophoresis (2-DE) and Matrix-Assisted Laser Desorption/Ionization (MALDI)-mass spectrometry were employed to investigate the differences of protein patterns in the hemolymph of the highly resistant, highly susceptible and near-isogenic silkworm strains after BmNPV was administrated to the larvae. A comparison between the proteomes of these three silkworm strains led us to identify two differentially expressed proteins, beta-N-acetylglucosaminidase 2 and aminoacylase. The expression levels of these proteins were higher in the BmNPV resistant strains.