Pupa

Diapause, an alternative developmental pathway characterized by changes in developmental timing and metabolism, is coordinated by molecular mechanisms that are not completely understood. MicroRNA (miRNA) mediated gene silencing is emerging as a key component of animal development and may have a significant role in initiating, maintaining, and terminating insect diapause. In the present study, we test this possibility by using high-throughput sequencing and qRT-PCR to discover diapause-related shifts in miRNA abundance in the flesh fly, Sarcophaga bullata. We identified ten evolutionarily conserved miRNAs that were differentially expressed in diapausing pupae compared to their nondiapausing counterparts. miR-289-5p and miR-1-3p were overexpressed in diapausing pupae and may be responsible for silencing expression of candidate genes during diapause. miR-9c-5p, miR-13b-3p, miR-31a-5p, miR-92b-3p, miR-275-3p, miR-276a-3p, miR-277-3p, and miR-305-5p were underexpressed in diapausing pupa...

Mosquitoes act as vectors of devastating pathogens and parasites, representing a key threat for millions of humans and animals worldwide. The control of mosquito-borne diseases is facing a number of crucial challenges, including the emergence of artemisinin and chloroquine resistance in Plasmodium parasites, as well as the presence of mosquito vectors resistant to synthetic and microbial pesticides. Therefore, eco-friendly tools are urgently required. Here, a synergic approach relying to nanotechnologies and biological control strategies is proposed. The marine environment is an outstanding reservoir of bioactive natural products, which have many applications against pests, parasites, and pathogens. We proposed a novel method of seaweed-mediated synthesis of silver nanoparticles (AgNP) using the spongeweed Codium tomentosum, acting as a reducing and capping agent. AgNP were characterized by UV-Vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, scanning electron micros...

G protein-coupled receptors (GPCRs) play key roles in a wide diversity of physiological processes and signalling pathways. The leucine-rich repeats containing GPCRs (LGRs) are a subfamily that is well-conserved throughout most metazoan phyla and have important regulatory roles in vertebrates. Here, we report on the critical role of Drosophila melanogaster LGR1, the fruit fly homologue of the vertebrate glycoprotein hormone receptors, in development as a factor involved in the regulation of pupariation. Transcript profiling revealed that lgr1 transcripts are most abundant in third instar larvae and adult flies. The tissues displaying the highest transcript levels were the hindgut, the rectum and the salivary glands. Knockdown using RNA interference (RNAi) demonstrated that white pupa formation was severely suppressed in D. melanogaster lgr1 RNAi larvae. Associated with this developmental defect was a reduced ecdysteroid titer, which is in line with significantly reduced transcript levels detected for the Halloween genes shadow (sad) and spookier (spok) in the third instar lgr1 RNAi larvae compared to the control condition.

Aedes aegypti, the major vector of dengue, breeds in domestic water containers. The development of immature mosquitoes in such containers is influenced by various environmental, ecological and socioeconomic factors. Urban and rural disparities in water storage practices and water source supply may affect mosquito immature abundance and, potentially, dengue risk. We evaluated the effect of water and container characteristics on A. aegypti immature abundance in urban and rural areas. Data were collected in the wet season of 2011 in central Colombia from 36 urban and 35 rural containers, which were either mosquito-positive or negative. Immature mosquitoes were identified to species. Data on water and container characteristics were collected from all containers. A total of 1452 Aedes pupae and larvae were collected of which 81% were A. aegypti and 19% A. fluviatilis. Aedes aegypti immatures were found in both urban and rural sites. However, the mean number of A. aegypti pupae was five t...

Effect of temperature on development of pink stem borer, Sesamia cretica Lederer, was studied at eight constant temperatures (15, 18, 20.5, 24, 27, 30, 34, and 38°C), a photoperiod of 16:8 (L:D) h, and 50-60% rela\tive humidity. The larvae of pink stem borer were reared on cutting stems of maize. The results showed that temperature had statistically significant effect on developmental times of the all developmental stages. The most commonly used six nonlinear models applied for modeling developmental rate of immature stages as a function of temperature. Evaluation of the models fit to data took place based on the coefficient of determination, residual sum of squires, adjusted coefficient of determination, and Akaike information criterion. Besides statistical criteria, biological significance was used to determine the best model. All the examined models statistically fit the data well. In addition, Briere-2 was selected as the best model considering biological significance of the est...

Correct and precise identification of mosquito vectors is important in many respects including development of vector control strategies. Conventional identification methods have limitations for sibling and closely related species of mosquitoes, stage and quality of the specimen used and this could be overcome by DNA-based identification methods using molecular markers such as nuclear ribosomal internal transcribed spacer (ITS) which do not demand intact or undamaged specimen. Genomic DNA is usually isolated from whole mosquito, legs, wings etc. Alternate sources for genomic DNA isolation such as eggshells, larval and pupal exuviae were explored in this study by amplifying the ITS markers. Standardization of genomic DNA extraction and ITS amplification were carried out with laboratory specimens. The same was applied to specimens collected from the field. The results show that PCR amenable genomic DNA could be isolated from fresh exuviae collected in the laboratory and not from older ...

We conducted this study to assess how well a pupal survey of dengue mosquito vectors, Aedes aegypti and Aedes albopictus, is able to target the most productive breeding sites. The study was carried out monthly during the rainy season (8 months) in 2008 in Cuba, Cebu City, Philippines. The hypotheses tested were: 1) most pupae of Ae. aegypti or Ae. albopictus were produced in a few types of breeding sites and 2) the most productive types of breeding sites for each species were the most abundant. Approximately 2,500 pupae were collected from 554 breeding sites in 279 houses. Thirty-eight point four percent of ten types of breeding sites were positive for Ae. aegypti, and 11.9% of nine types of sites were positive for Ae. albopictus. Plastic drums (40.2%), metal drums (29.6%), and plastic containers (10.5%) were the key sites for Ae. aegypti pupae, whereas bamboo stumps (28.5%), plastic drums (21.1%), and rubber tires (19.1%) were the key sites for Ae. albopictus. The most productive b...

Rapid and efficient DNA-based tools are recommended for the evaluation of the insect biodiversity of high-altitude streams. In the present study, focused principally on larvae of the genus Diamesa Meigen 1835 (Diptera: Chironomidae), the congruence between morphological/molecular delimitation of species as well as performances in taxonomic assignments were evaluated. A fragment of the mitochondrial cox1 gene was obtained from 112 larvae, pupae and adults (Diamesinae, Orthocladiinae and Tanypodinae) that were collected in different mountain regions of the Alps and Apennines. On the basis of morphological characters 102 specimens were attributed to 16 species, and the remaining ten specimens were identified to the genus level. Molecular species delimitation was performed using: i) distance-based Automatic Barcode Gap Discovery (ABGD), with no a priori assumptions on species identification; and ii) coalescent tree-based approaches as the Generalized Mixed Yule Coalescent model, its Bay...

Twenty-nine taxa of Simulium were identified amongst 527 collections of larvae and pupae from untreated rivers and streams in Liberia (362 collections in 1967-71 & 1989), Togo (125 in 1979-81), Benin (35 in 1979-81) and Ghana (5 in 1980-81). Presence or absence of associations between different taxa were used to group them into six clusters using Ward agglomerative hierarchical cluster analysis. Environmental data associated with the pre-imaginal habitats were then analysed in relation to the six clusters by one way ANOVA. The results revealed significant effects in determining the clusters of maximum river width (all P<0.001 unless stated otherwise), water temperature, dry bulb air temperature, relative humidity, altitude, type of water (on a range from trickle to large river), water level, slope, current, vegetation, light conditions, discharge, length of breeding area, environs, terrain, river bed type (P<0.01), and the supports to which the insects were attached (P<0.01...

Polypedilum nubifer (Skuse, 1889), originally described from Australia, is an apparently widespread species of Chironomidae (Diptera) that can attain nuisance densities in some eutrophic water bodies. Appropriate management depends upon the identity and ability to distinguish from potential cryptic taxa. A morphological study of larvae, pupae and adults of both sexes confirmed P. nubifer as widely distributed and frequently abundant, but also revealed two previously cryptic species of limited distribution in northern Australia. These species are described as new and illustrated in all stages here. Polypedilum quasinubifer Cranston sp. n. is described from north-west Queensland, Australia and also from Thailand and Singapore. Polypedilum paranubifer Cranston sp. n. is known only from retention ponds of a uranium mine in Northern Territory, Australia. Unusual morphological features of P. nubifer including alternate Lauterborn organs on the larval antenna, cephalic tubules on the pupa ...

Seasonal polyphenism, in which different forms of a species are produced at different times of the year, is a common form of phenotypic plasticity among insects. Here I show that the production of dark fifth-instar caterpillars of the eastern black swallowtail butterfly, Papilio polyxenes, is a seasonal polyphenism, with larvae reared on autumnal conditions being significantly darker than larvae reared on midsummer conditions. Both rearing photoperiod and temperature were found to have individual and synergistic effects on larval darkness. Genetic analysis of variation among full-sibling families reared on combinations of two different temperatures and photoperiods is consistent with the hypothesis that variation in darkness is heritable. In addition, the genetic correlation in larval darkness across midsummer and autumnal environments is not different from zero, suggesting that differential gene expression is responsible for the increase in larval darkness in the autumn. The relatively dark autumnal form was found to have a higher body temperature in sunlight than did the lighter midsummer form, and small differences in temperature were found to increase larval growth rate. These results suggest that this genetically based seasonal polyphenism in larval color has evolved in part to increase larval growth rates in the autumn.

We used classical culture techniques to explore gut bacteria and changes associated with dietary change in the highly polyphagous, tropical caterpillar Automeris zugana (Saturniidae). Fifty-five third instar wild-caught sibs feeding on Annona purpurea (Annonaceae) in the Area de Conservación Guanacaste (ACG) in northwestern Costa Rica were divided into eight groups. Each of seven groups was reared to the ultimate instar on another species of food plant normally used by A. zugana. Some pupae were also analyzed for the presence of bacteria. Aerobic bacterial cultures were obtained from all 33 caterpillar guts and the eight pupae inventoried. There was no clear pattern in species composition of cultivated bacteria among the eight diets, and each caterpillar on a given food plant carried only a small fraction of the total set of species isolated from the set of caterpillars feeding on that food plant. Taken as a whole, the larvae and pupae contained 22 species of cultivable bacteria in ...

Both phytophagous and parasitic insects deposit oviposition-marking pheromones (OMPs) following oviposition that function to inform conspecifics of a previously utilized host of reduced suitability. The blueberry maggot fly, Rhagoletis mendax Curran (Diptera: Tephritidae), deposits eggs individually into blueberries and then marks the fruit surface with an OMP which reduces acceptance of fruit for oviposition by conspecifics. Diachasma alloeum (Muesebeck) (Hymenoptera:

We developed a bioassay to measure the flying power of drone, in order to determine which drones could reach a drone congregation area. A wind tunnel was used to test unparasitized drones and drones slightly parasitized by one or two mites during pupal development, and counts were made of the number of spermatozoa that they produced. Drones parasitized with one mite flew as long as control drones (x= 6'55" and 6'48", respectively, P = 0.512); however, those that had been infested by two mites flew significantly less (x= 2'16", P<0.001). There was a significant positive correlation (P<0.01) between flight duration and the number of spermatozoa per drone in control group (r = 0.53), and in both the one mite (r = 0.43) and two mite (r = 0.54) groups. Drones infested during development with one or two mites produced 24 and 45% fewer sperm, respectively.

The routine colonization of Anopheles maculatus, a reputed malaria vector from Central Java, is described. The strain is free mating and long lived in the laboratory. This species will readily bloodfeed on small rodents and artificial membrane systems. Either natural or controlled temperatures, humidity, and lighting provide acceptable conditions for continuous rearing. A simple larval diet incorporating a 10:4 powdered mixture of dried beef and rice hulls proved acceptable. Using a variety of simple tools and procedures, this colony strain appears readily adaptable to rearing under most laboratory conditions. This appears to be the first report of continuous colonization using a free-mating strain of An. maculatus. Using this simple, relatively inexpensive method of mass colonization adds to the short list of acceptable laboratory populations used in the routine production of human-infecting plasmodia.