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PubMed1975

Insect immunity. 11. Simultaneous induction of antibacterial activity and selection synthesis of some hemolymph proteins in diapausing pupae of Hyalophora cecropia and Samia cynthia.

We have previously shown that pupae of the giant silkmoth Samia cynthia have a humoral antibacterial activity, which was induced by viable, nonpathogenic gram-negative bacteria (H.G. Boman et al., 1974). We show here that this activity was formed simultaneously with a selective incorporation of amino acids into eight polypeptide chains characterized by their electrophoretic behavior. If actinomycin D or cycloheximide were given at an early time, no antibacterial activity was found. If the inhibitors were given at the time of maximum activity, there was no effect with actinomycin D but a rapid decrease of the activity in the case of cycloheximide. The results imply that the messenger ribonucleic acid was stable, but that at least one protein component was turning over. Hemolymph from immunized pupae of another giant silkmoth, Hyalophora cecropia, was fractionated by ammonium sulfate precipitation. This procedure, together with the isotope distribution after co-electrophoresis in polyarylamide gels, was used for comparing the response to injury and to different infections. Almost identical polypeptide patterns were obtained as a response to an infection with either viable Enterobacter cloacae or Bacillus subtilis. These patterns differed both qualitatively and quantitatively from the injury effect created by an injection as such. There was only a low antibacterial activity in each of the four fractions obtained by ammonium sulfate precipitation. However, a combination of three fractions restored a high killing activity. Fractionation of hemolymph from untreated pupae provided evidence for at least one preexisting factor which stimulated the killing of Escherichia coli. The osmotic pressure of the bacteria contributed to the antibacterial activity towards E. coli, but not towards B. subtitlis. The killing of E. coli was inhibited by liped A and, to a lesser extent, by an inhibitor of proteolytic enzymes. The similarities and differences with the mammalian complement system are discussed.

Animals↗
PubMed2006

Sample sizes for identifying the key types of container occupied by dengue-vector pupae: the use of entropy in analyses of compositional data.

A method has been developed for estimating the sample sizes needed to identify categories that comprise a large proportion of a compositional data-set. The method is to be used in the design of surveys of mosquito pupae, for identifying the key container types from which the majority of adult dengue vectors emerge. Although a finite-population correction was devised for estimating the mean of a negative binomial distribution, other complications of parametric approaches make them unlikely to yield methods simple enough to be practically applicable. The Shannon-Wiener index was therefore investigated as a more useful alternative, at the cost of theoretical generalizability, in an approach based on re-sampling methods in conjunction with the use of entropy. This index can be used to summarize the degree to which pupae are either concentrated in a few container types, or dispersed among many. An empirical relationship between the index and the repeatability of surveys of differing sample sizes was observed. A step-wise rule, based on the entropy of the cumulative data, was devised for determining the sample size, in terms of the number of houses positive for pupae, at which a pupal survey might reasonably be stopped.

Aedes↗
PubMed1996

Effect of engineering Hsp70 copy number on Hsp70 expression and tolerance of ecologically relevant heat shock in larvae and pupae of Drosophila melanogaster.

To determine how the accumulation of the major Drosophila melanogaster heat-shock protein, Hsp70, affects inducible thermotolerance in larvae and pupae, we have compared two sister strains generated by site-specific homologus recombination. One strain carried 12 extra copies of the Hsp70 gene at a single insertion site (extra-copy strain) and the other carried remnants of the transgene construct but lacked the extra copies of Hsp70 (excision strain). Hsp70 levels in whole-body lysates of larvae and pupae were measured by ELISA with an Hsp70-specific antibody. In both extra-copy and excision strains, Hsp70 was undetectable prior to heat shock. Hsp70 concentrations were higher in the extra-copy strain than in the excision strain at most time points during and after heat shock. Pretreatment (i.e. exposure to 36 degrees C before heat shock) significantly improved thermotolerance, and this improvement was greater and more rapid in larvae and pupae of the extra-copy strain than in those of the excision strain. The experimental conditions resemble thermal regimes actually experienced by Drosophila in the field. Thus, these findings represent the best evidence to date that the amount of a heat-shock protein affects the fitness of a complex animal in the wild.

Animals↗
PubMed1999

Regulation of swimming in the Culex pipiens (Diptera, Culicidae) pupa: kinematics and locomotory trajectories.

High-speed videography was used to investigate swimming kinematics and locomotory trajectories during escape responses in the pupa of Culex pipiens (Diptera, Culicidae). The pupa can perform straight-line motion despite undergoing backward and forward somersaults through 400 degrees at a rate of 20 s(-)(1). For linear motion to occur, the effective stroke of the abdomen must be delivered during the part of each cycle when the body is facing forwards, allowing passive rotation to carry the body round to the correct position for the start of the next half-stroke. On-off motion of the abdomen is regulated by a click mechanism based on stressed integumentary plates which buckle at strategic points in the cycle. The importance of self-inertia as a component in positional regulation was demonstrated by increasing the kinematic viscosity of the medium: this retarded rotation and resulted in somersaulting on the spot. Whereas normally the pupa invariably directs its swimming downwards, reversal of the usual light gradient produced upward paths as well. It is concluded that straight-line motion is generated automatically by the locomotory machine without any need for sensory feedback about body orientation during each cycle, but that swimming direction can be influenced by sensory cues such as light.

Animals↗
PubMed2001

The vortex wake of the free-swimming larva and pupa of Culex pipiens (Diptera).

The kinematics and hydrodynamics of free-swimming pupal and larval (final-instar) culicids were investigated using videography and a simple wake-visualisation technique (dyes). In both cases, swimming is based on a technique of high-amplitude, side-to-side (larva) or up-and-down (pupa) bending of the body. The pupa possesses a pair of plate-like abdominal paddles; the larval abdominal paddle consists of a fan of closely spaced bristles which, at the Reynolds numbers involved, behaves like a continuous surface. Wake visualisation showed that each half-stroke of the swimming cycle produces a discrete ring vortex that is convected away from the body. Consecutive vortices are produced first to one side then to the other of the mean swimming path, the convection axis being inclined at approximately 25 degrees away from dead aft. Pupal and larval culicids therefore resemble fish in using the momentum injected into the water to generate thrust. Preliminary calculations for the pupa suggest that each vortex contains sufficient momentum to account for that added to the body with each half-stroke. The possibility is discussed that the side-to-side flexural technique may allow an interaction between body and tail flows in the production of vorticity.

Animals↗
PubMed2005

Investigation of relationships between Aedes aegypti egg, larvae, pupae, and adult density indices where their main breeding sites were located indoors.

Aedes aegypti (L.) density indices obtained in a dengue fever (DF) endemic area were compared. One hundred and twenty premises, in an urban area of Colombia where dengue type-1 and type-2 virus cocirculated, were randomly selected and sampled for 7 months. The geometric mean monthly numbers (density index, DI) of Ae. aegypti eggs (ODI), 4th instar larvae (LDI), pupae (PDI), and adults (ADI) were calculated based on the use of ovitraps, nets, and manual aspirators, respectively. A negative temporal correlation was observed between the LDI and the ODI (r = -0.83, df = 5, and P < 0.01). Positive temporal correlations were only observed between the LDI and the PDI (r = 0.90, df = 5, and P < 00.5) and the Breteau and House indices (r = 0.86, df = 5, and P < 0.01). No other correlations were found between these indices and any of the other density indices or the incidence of suspected DF cases in residents, the temperature, the rainfall, or seasonal fluctuations. Our results were, therefore, probably due to the most productive Ae. aegypti breeding sites (large water containers) being located indoors within this study area. The number of adult female Ae. aegypti/person (n = 0.5) and pupae/person (n = 11) in our study area were lower and dramatically higher than the transmission thresholds previously reported for adult and pupae, respectively. Because there were confirmed DF cases during the study period, the transmission threshold based on the Ae. aegypti pupae was clearly more reliable. We found that the mean ovitrap premise index (OPI) was 98.2% during this study and that the mean larval (L-4th instars) premise index (LPI) was 59.2%, and therefore we suggest that the OPI and LPI would be more sensitive methods to gauge the effectiveness of A. aegypti control programs.

Aedes↗
PubMed1957

Toxicity of some newer insecticides to larvae and pupae of Culex fatigans Weidemann.

The toxicity of DDT, dieldrin, endrin, malathion, parathion, Systox and P4 to larvae and pupae of Culex fatigans has been studied. Two colonies of mosquitos were maintained for the purpose-one at 88 degrees +/-2 degrees F (31.1 degrees +/-1.1 degrees C) and 70%-80% relative humidity, and the other at 72 degrees +/-3 degrees F (22.2 degrees +/-1.7 degrees C) and 45%-55% relative humidity.For test purposes 25 larvae or pupae were exposed to acetone solutions of insecticides in 25 ml of water, and mortality counts were taken at the end of 24 and 48 hours. The percent mortality was plotted on a logarithmic dosage-probit paper and the LC(50) (the concentration required to kill 50%) was read from the slopes obtained. The confidence limits for LC(50) and the slope functions were worked out statistically.Of the insecticides tested, parathion was found to be the most toxic to larvae. The others in their order of toxicity were endrin, dieldrin, DDT, malathion, P4 and Systox. The LC(50) at 24 hours ranged from 0.003 parts per million (p.p.m.) for parathion to 0.18 for Systox. The third-instar larvae were more susceptible to insecticides than those in the fourth-instar stage.Pupae were more resistant to insecticides than larvae. Parathion and endrin gave LC(50)'s of 0.034 and 0.04 respectively. P4 and malathion gave very poor results. Dieldrin and Systox acted slowly on pupae, and the 48-hour mortality is given for these. DDT showed extremely low toxicity to pupae.DDT, dieldrin, endrin and parathion were more toxic to larvae reared and tested at the lower temperature than to those reared and tested at the higher. The temperature after exposure to DDT altered toxicity to a greater extent than the temperature at which the larvae had been reared.

Animals↗
PubMed2005

Communal use of integumental wounds in honey bee (Apis mellifera) pupae multiply infested by the ectoparasitic mite Varroa destructor.

The ectoparasitic bee mite, Varroa destructor, is highly adapted to its natural and adopted honey bee hosts, Apis cerana and Apis mellifera. Adult females perforate the integument of bee pupae in such a way that they and their progeny can feed. We examined the wounds that founder females made, and usually found one, and rarely up to three, integumental wounds on pupae of A. mellifera multiply infested by V. destructor. The punctures were mainly on the 2nd abdominal sternite of the host. These perforations are used repeatedly as feeding sites by these hemolymph-sucking mites and by their progeny. The diameter of the wounds increased during pupal development. In brood cells containing 4-5 invading female mites and their progeny, healing of the wound is delayed, normally occurring just before the imaginal moult of the bee pupa. These wounds are subject to microbial infections, and they are relevant to the evolution of behavioral traits in these parasitic mites and their relations to host bees.

Animals↗
PubMed2003

[Abundance indices of larvae and pupae of Aedes aegypti (Diptera: Culicidae) in Córdoba City].

BACKGROUND: In 1995, Aedes aegypti, vector of the Flavivirus Dengue and Yellow Fever, reached the province. Therefore, it is necessary to quantify the vector abundance in order to determine the level of risk and to evaluate the prevention and control actions implemented. OBJECTIVES: To determine the vector abundance breeding place preferences. MATERIALS AND METHODS: Surveys of Ae. aegypti larva-pupa-infested containers were conducted in six neighborhoods of Córdoba city in order to determine the House, Container, and Breteau Indices. RESULTS: Larvae and pupae of Ae. aegypti were found in a great variety of containers, most of which had been discarded. The House Index obtained was 47 %, while the percentage of water-holding containers examined that had larva or pupa was 24 %. The Breteau Index was estimated at 106 %. CONCLUSIONS: According to the indices estimated, Córdoba city is in a risk situation if any infected person with any dengue virus enters our city. Therefore, it is necessary to intensify the measures, mainly the sanitary education, vector surveillance and control, in order to prevent any form of dengue in the region.

Aedes↗
PubMed1986

Efficacy of cypermethrin for the control of mosquito larvae and pupae, and impact on non-target organisms, including fish.

In laboratory tests, cypermethrin was highly toxic to mosquito larvae and pupae. It was more toxic at low temperatures after a 24 hr exposure. Larvae of Aedes stimulans were less susceptible than Culex restuans. Technical cypermethrin was more toxic than an emulsifiable concentrate formulation. In outdoor simulated pools cypermethrin 40% EC was consistently effective against larvae and pupae of Ae. stimulans at 10 g AI/ha and Culex spp. at 50 g AI/ha. When stickleback fish were tested, no mortality occurred at the lowest effective dosage in each trial. The residual toxicity of cypermethrin increased with dosage and was much higher in a test at 8 degrees C than at 20 degrees C. In natural snowmelt pools, cypermethrin at 20 g AI/ha provided 92-100% control of Aedes spp. larvae and pupae by 7 days after treatment. Non-target amphipod, anostracan, cladoceran and insect populations were usually reduced 80-100% while copepods, ostracods and hydracarinid mites were generally less affected. No significant mortality of caged stickleback fish occurred in these pools.

Animals↗
PubMed1994

New techniques for rearing black flies from pupae (Diptera: Simuliidae).

Simple techniques are described for collecting black fly pupae from streams using plastic strips and for rearing large numbers of adult black flies in inexpensive enclosures made of chicken wire and cloth mesh netting, and 2 methods are described for rearing adult black flies individually from pupae. The first method of rearing individual black flies uses 1.5-ml microcentrifuge tubules and the second uses easy-to-construct rearing chambers that provide moisture for the developing pupa and water for the adult to imbibe. Instructions for assembly are provided. Specimens obtained using these rearing chambers are of museum quality.

Animals↗
PubMed1994

Afrotropical Culicoides: description and comparison of the pupae of seven species of the Similis supergroup (Diptera: Ceratopogonidae).

The pupae of Culicoides exspectator Clastrier, C. kobae Cornet & Chateau. C. micheli Cornet & Chateau, C. olyslageri Kremer & Nevill. C. ravus De Meillon and C. (Synhelea) tropicalis Kieffer are described and illustrated for the first time, and the pupa of C. similis Carter, Ingram & Macfie is redescribed. All seven species belong to the Similis supergroup; the immature stages of six are found in ground waters while those of C. olyslageri occur in tree holes. A concise key is presented for differentiating the seven species. Short notes on their larval habitats are given. Reference is made to C. accraensis Carter, Ingram & Macfie, also found in tree holes. Available data suggest that pupal morphology can be used to differentiate closely related species; group characters also exist for those species within the Similis supergroup which can be used to separate ground-water species from those utilizing tree holes. Indications are that the pupae are of limited help in resolving the systematic subdivision of the Similis supergroup.

Animals↗
PubMed1975

[Hemocytic reactions in lepidoptera pupae. Intensity of the reaction as a function of its localization in Pieris brassicae].

Quantitative studies were made on cellular capsules formed in diapause pupae of Pieris brassicae. A simple procedure is to implant pieces of nylon thread in hemocoele. Differences have been noticed in the vigour of the reaction at different regions of the pupae : the haemocytic reaction was less vigorous in the central region of pupae than in the distal ones.

Animals↗
PubMed2001

Development of the sensory system in larvae and pupae of Chaoborus crystallinus (DeGeer, 1776; Diptera, Chaoboridae): sensory cells, nerves and ganglia of the tail region.

Using various microscopical techniques, we have studied changes in the sensory equipment and architecture of the peripheral nervous system (PNS) around the first metamorphic molt from larva to pupa in the phantom midge Chaoborus. The transparent larvae and pupae of this dipteran with ancestral features allow us to investigate sensilla and their central projections from whole-mount preparations of complete groups of segments. Each sensillum on the posterior larval and pupal segments was identified using its external shape and position, and the morphology of the abdominal ganglia and segmental nerves was investigated. In addition, retrograde fills with the carbocyanine dye DiI were used to trace the axonal paths of most of the extero- and proprioreceptors. These findings were combined to produce maps of the sensory elements of larval and pupal abdomens that were analyzed at three levels: seriality (homonomy), ontogenetic changes of individual sensilla, and homology of the PNS between different species. Comparison of different segments shows for both stages that primarily there is a homonomous basic design of the PNS, but segment-specific modifications are evident in segments 8-10. Comparison of corresponding larval and pupal segments shows that many sensilla retain their internal structure and axonal projections. However, their external cuticular parts are changed in relation to the different life habits of larvae and pupae. Furthermore, some sensilla are completely reduced during the pupal molt, especially those of the tenth segment which appears as a distinct larval structure (caenogenesis). Comparison between species indicates that despite the varying types of sensilla their basic segmental arrangement and their axonal trajectories are conserved.

Animals↗
PubMed1985

Accelerated aging of the insect Drosophila melanogaster by gamma irradiations of pupae.

The purpose of this study was to induce accelerated aging in the fly Drosophila melanogaster (Diptera) using free radicals, and to distinguish aging from radiation syndrome. After irradiation of 12 to 24-hour-old pupae with does of 500-2500 rad, very similar fly emergence rates were observed in control and treated populations. The life span was shortened and dose-related in mixed populations and in both virgin and mated male and female populations. Males were more radiosensitive than females. In the 500-2500 rad dose range, all the populations tested were fertile after irradiation. Young treated as pupae and control adults both displayed similar mating ability and negative geotactic responses. Their response levels decreased with time, but the decline was faster and dose-related in the treated animals. This study demonstrates that irradiation of 12 to 24-hour-old pupae with doses of 500-2500 rad induces accelerated aging, without causing serious biological damage. For example, after an irradiation with 1750 rad, the "lethal time" (LT) 50 dropped by about 30%.

Aging↗
PubMed2000

Complete sequence of the mitochondrial DNA of the primitive opisthobranch gastropod Pupa strigosa: systematic implication of the genome organization.

The complete sequence (14,189 bp) of the mitochondrial DNA of the opisthobranch gastropod Pupa strigosa was determined. The genome contains 13 protein, 2 rRNA, and 22 tRNA genes typical of metazoan mtDNA. The Pupa mitochondrial genome is highly compact and shows the following unusual features, like pulmonate land snails: (1) extremely small genome size, (2) absence of lengthy noncoding regions (with the largest intergenic spacer being only 46 nt), (3) size reduction of encoded genes, and (4) many overlapping genes. Several tRNA genes exhibit bizarre secondary structures with reduced T or D stems, and many tRNA genes have unstable acceptor stems that might be corrected by posttranscriptional RNA editing. The Pupa mitochondrial gene arrangement is almost identical to those of pulmonate land snails but is radically divergent from those of the prosobranch gastropod Littorina saxatilis and other molluscs. Our finding that the unique gene arrangement and highly compact genome organization are shared between opisthobranch and pulmonate gastropods strongly suggests their close phylogenetic affinity.

Amino Acid Sequence↗
PubMed1992

Induction of a novel protein kinase in pupae of the silkworm Bombyx mori after infection with nuclear polyhedrosis virus.

Protein kinases induced by Bombyx mori nuclear polyhedrosis virus in pupae of the silkworm B. mori were examined by activity gel analysis using phosvitin as a protein substrate. The method involved PAGE of the soluble fraction from pupae under native conditions and in the presence of SDS, followed by in situ renaturation of proteins and recovery of protein kinase activity in the intact gel. A novel protein kinase able to phosphorylate phosvitin was detected in the infected pupae from 2 days post-infection. This enzyme was not present in uninfected silkworms at any stage of the pupal period. The novel kinase activity was found by SDS-PAGE to be associated with a single polypeptide with an apparent M(r) of 50K. However, on electrophoresis under native conditions its activity was associated with a set of polypeptides with similar but not identical electrophoretic mobilities. Microheterogeneity of the catalytically active polypeptides suggests that the virus-induced protein kinase undergoes post-translational modification during the course of infection.

Animals↗
PubMed1982

Insect immunity: isolation and structure of cecropins B and D from pupae of the Chinese oak silk moth, Antheraea pernyi.

The immune system in the Chinese oak silk moth, Antheraea pernyi, has been compared with that of the Cecropia moth which has been characterized earlier. Antibacterial activity against Escherichia coli was induced in diapausing pupae by injection of viable E. coli or Enterobacter cloacae. The activity reached a maximum on day 7-8 after which the response gradually declined. The pupae produced a set of immune proteins with P4 and P5 as major labelled components similar to that earlier found in Cecropia. The major antibacterial factor in A. pernyi was cecropin D. A procedure is described for the isolation of cecropin B and D, which is in principle similar to the one used for the isolation of the corresponding cecropins from Cecropia pupae. Amino acid sequence analyses of the A. pernyi cecropins show the D form to contain 36 amino acid residues and that both cecropins have blocked C-termini. The general structure of cecropins having a charged N-terminal region (residues 1-21) followed by a long hydrophobic stretch (residues 22-32) is well conserved. Cecropin B and D from A. pernyi differ from the corresponding proteins in Cecropia by four and three conservative amino acid replacements, respectively. The homology between the cecropins from the two insects suggests that they orginate from a single ancestral gene. The antibacterial activity was tested against nine different bacterial species. Evolutionary aspects of the cecropins are discussed.

Amino Acids↗