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Effect of ten chlorophytes on larval survival, development and adult body size of the mosquito Aedes aegypti.

The effect of ten microalgal chlorophytes isolated from mosquito breeding containers on the survival, larval development and adult body size of the mosquito Aedes aegypti was investigated. All larvae fed with six of the microalgal isolates died after 7 days. These isolates were found to be resistant to digestion by mosquito larvae. Delayed pupation and body size reduction of the mosquitos fed with Chlorococcum UMACC 218 and Scenedesmus UMACC 220 were observed. In contrast, larvae fed with Ankistrodesmus convolutus UMACC 101 and Chlorococcum UMACC 213 were bigger in size than those fed with normal insectory feed. The present study showed that microalgal chlorophytes have the potential to be used as larvicidal agents for mosquitos.

Aedes↗

Soluble and particle-bound trehalase in the silk glands during larval-pupal development of the silkworm, Bombyx mori.

Trehalase localization in silk glands of the silkworm, Bombyx mori has been studied during larval-pupal development. Subcellular distribution of the silk gland trehalase depends upon larval-pupal development. The activity increases in the soluble fraction with a concomitant decrease in the particle-bound fraction during larval-pupal development. The pH-optimum value of trehalase activity in the particle-bound fraction changes from 6.5 to 6.0, and in the soluble fraction from 5.5 to 4.5 in the course of the silk gland degeneration during metamorphosis.

Animals↗

Neurogenesis in the thoracic neuromeres of two crustaceans with different types of metamorphic development

The mode of embryonic and larval development and the ethology of metamorphosis in the spider crab and the American lobster are very different, and we took advantage of this to compare neuronal development in the two species. The goals of this study were to discover whether the differences in the maturation of the neuromuscular system in the pereopods and the metamorphic changes of motor behavior between the two species are reflected at the level of the developing nervous system ('neurometamorphosis'). Furthermore, we wanted to broaden our understanding of the mechanisms that govern neuronal development in arthropods. Proliferation of neuronal stem cells in thoracic neuromeres 4-8 of the lobster Homarus americanus and the crab Hyas araneus was monitored over the course of embryonic and larval development using the in vivo incorporation of bromodeoxyuridine (BrdU). Neuropil structure was visualized using an antibody against Drosophila synapsin. While proliferation of neuronal precursors has ceased when embryogenesis is 80 % complete (E80%) in the lobster thoracic neuromeres, proliferation of neuroblasts in the crab persists throughout embryonic development and into larval life. The divergent temporal patterns of neurogenesis in the two crustacean species can be correlated with differences in larval life style and in the degree of maturation of the thoracic legs during metamorphic development. Several unusual aspects of neurogenesis reported here distinguish these crustaceans from other arthropods. Lobsters apparently lack a postembryonic period of proliferation in the thoracic neuromeres despite the metamorphic remodeling that takes place in the larval stages. In contrast, an increase in mitotic activity towards the end of embryonic development is found in crabs, and neuroblast proliferation persists throughout the process of hatching into the larval stages. In both E20% lobster embryos and mid-embryonic crabs, expression of engrailed was found in a corresponding set of neurons and putative glial cells at the posterior neuromere border, suggesting that these cells have acquired similar specific identities and might, therefore, be homologous. None of the BrdU-labeled neuroblasts (typically 6-8 per hemineuromere over a long period of embryogenesis) was positive for engrailed at this and subsequent stages. Our findings are discussed in relation to the spatial and temporal patterns of neurogenesis in insects.

Journal Article↗

Energy, development and fitness in Drosophila melanogaster.

Perturbation of energy metabolism via NAD supplementation modifies the expression of larval development time, a genetically based trait relevant to development and whole-organism fitness in Drosophila melanogaster. The results suggest a link between energy metabolism and gene expression. Supplemental NAD significantly decreased larval development time for isofemale strains chosen on the basis of development time differences. This effect did not represent a trade-off with larval survival, and was not related to the effects of supplemental glucose. The isofemale strains were assayed by using a method derived from Warburg's manometric procedures which measured changes in the proportions of ATP, ADP and AMP in larval homogenates. Supplemental NAD appeared to increase the proportion of ATP and the [ATP]/[ADP] ratio in vitro and during larval development. The decrease in larval development time mediated by NAD was attributed to supplemental NAD modifying the NAD pool beyond levels established during development, directly altering electron transport chain activity, ATP, ADP and AMP levels, and possibly gene expression.

Adenine Nucleotides↗

[Possibilities of synchronizing the growth of Chironomus thummi Kieff. larvae].

The conditions for uniform synchronous development of groups of the IV instar larvae of Ch. thummi and the possibility of using the morphology of imaginal discs as a criterion of temporal uniformity (synchrony) of larval development were investigated. The degree of synchrony of larval development was shown to depend strongly on the population density. Its increase led to desynchronization and decrease in the rate of larval development. The size and morphology of imaginal discs used as criteria of developmental phases of the IV instar larvae varied at the "red head" phase. the size of imaginal discs depended on the rate of larval development and population density. No distinct correlation was found between the sex of larvae and the size of imaginal discs in them. A conclusion is drawn that the size of imaginal discs cannot serve as a criterion of temporal uniformity of larval development.

Animals↗

Embryotoxic effects of environmental chemicals: tests with the South African clawed toad (Xenopus laevis).

In the course of the investigations reported below, it was shown that p-chloroaniline has a lethal effect on the embryos of Xenopus laevis at a concentration of 100 ppm and is development inhibiting (teratogenic) at concentrations of 1 and 10 ppm, respectively. In the case of aniline, a significant development-inhibiting effect was observed at a concentration as low as 1 ppm. A toxic effect was caused by concentrations between 30 and 40 ppm during embryogenesis and by concentrations above 40 ppm during larval development. A very conspicuous finding was an inhibiting effect of 20 to 40 ppm aniline on pigmentation during embryogenesis and of a concentration as low as 1 ppm on the body size of the young toads. In the case of potassium dichromate, it was possible to barely detect a weak development-inhibiting effect during embryogenesis but no development-retarding effect during larval development. Toxic effects of potassium dichromate occurred during embryogenesis at concentrations of 5 and 7.5 ppm and during the larval development at concentrations above 10 ppm. Sodium dodecylbenzenesulfonic acid at a concentration of 50 ppm was found to have such a strong embryolethal effect that 80% of the eggs showed no cell division at all and the remaining 20% developed to only the bicellular stage. A teratogenic effect of this substance was not observed. Phenol, too, was found to be toxic at a concentration of 50 ppm; in contrast to sodium dodecylbenzenesulfonic acid, however, it did not show any lethal effect on the embryos but it did on the tadpoles, mainly in the first stages of larval development. Lower concentrations of phenol (5 and 10 ppm) had a nonsignificant inhibiting effect on the growth of the larvae. A teratogenic effect of phenol was not detected.

Aniline Compounds↗

The molecular mechanisms of larval cestode development: first steps into an unknown world.

Several hundred million years ago, the free-living ancestors of all extant helminth parasites decided to colonize entirely new habitats, the bodies of other metazoan animals. As a consequence of the resulting adaptation processes, they evolved highly complex life-cycles in which many developmental transitions were initiated and controlled by host-derived signals. Understanding the molecular basis of the original developmental mechanisms, and the modifications that occurred during co-evolution with the host, is not only fundamental to our understanding of parasitism but also highly relevant for the design of anti-parasitic drugs and vaccines. In the past several years, molecular investigations on parasitic nematode and trematode development have made considerable progress and, supported by respective genome sequencing projects and emerging methods of genetic manipulation, will be a flourishing field in the years to come. We consider it time that corresponding studies are also pushed for the third large group of parasitic helminths, the cestodes. Here, we review the first experimental steps into that area, which have been undertaken recently. We report on cestode genomics, the identification of signaling factors associated with larval development, and the establishment as well as improvement of in vitro cultivation systems by which cestode life-cycles can be studied in the laboratory.

Animals↗

Mutants altering coordinate synthesis of specific myosins during nematode muscle development.

Mutations in the unc-52 gene on linkage group II retard the construction of body-wall muscle sarcomeres during larval development in the nematode Caenorhabditis elegans. Unc-52 mutants show decreased accumulation of myosin heavy chains relative to other polypeptides during larval development, correlating with the structural retardation. Pulse radiolabeling experiments show that decreased synthesis of specific body-wall myosin heavy chains that are encoded by the unc-54 gene on linkage group I is responsible for the defective myosin accumulation. In the wild type, a constant ratio of the synthesis of the unc-54-coded myosin B to myosin A, about 2:1, is maintained during the larval stages in which the synthesis of both myosins increases exponentially and rapid sarcomere growth and addition ensues. During the first 26 hr of larval development, before any structural or behavioral effects of unc-52 mutations are apparent, the synthesis of myosin heavy chains is also normal. By 38 hr, decreased synthesis of myosin B is detected in the unc-52 mutant SU200, when sarcomere growth slows considerably. The effects of mutation in the unc-52 locus are trans acting upon the synthesis of unc-54-coded myosin in a specific set of muscle cells during a defined period of larval development.

Animals↗

Delayed insemination results in embryo mortality in a brooding ascidian.

We explored the effects of temporal variation in sperm availability on fertilization and subsequent larval development in the colonial ascidian Botryllus schlosseri, a brooding hermaphrodite that has a sexual cycle linked to an asexual zooid replacement cycle. We developed a method to quantify the timing of events early in this cycle, and then isolated colonies before the start of the cycle and inseminated them at various times. Colony-wide fertilization levels (assayed by early cleavage) increased from zero to 100% during the period when the siphons of a new generation of zooids were first opening, and remained high for 24 h before slowly declining over the next 48 h. Because embryos are brooded until just before the zooids degenerate at the end of a cycle, delayed fertilization might also affect whether embryos can complete development within the cycle. Consequently, we also determined the effect of delayed insemination on successful embryo development through larval release and metamorphosis. When fertilization was delayed beyond the completion of siphon opening, there was an exponential decline in the percentage of eggs that ultimately produced a metamorphosed larva at the end of the cycle. Thus, even though the majority of oocytes can be fertilized when insemination is delayed for up to 48 h, the resulting embryos cannot complete development before the brooding zooids degenerate.

Animals↗

Comparison of six in vitro tests in determining benzimidazole and levamisole resistance in Haemonchus contortus and Ostertagia circumcincta of sheep.

Six in vitro methods for the detection of anthelmintic resistance were compared using benzimidazole/levamisole-resistant Haemonchus contortus and benzimidazole/levamisole/ivermectin-resistant Ostertagia circumcincta as well as susceptible strains of both parasite species. The degree of resistance to thiabendazole and levamisole was compared by (1) an egg hatch assay, (2) an egg hatch paralysis assay, (3) a larval development assay, (4) a larval paralysis assay (5) a larval paralysis assay with physostigmine and (6) larval micromotility assay. The degree of resistance for each assay was expressed as resistance factor--RF. For the detection of thiabendazole and levamisole resistance, the larval development test was observed as the most sensitive to measure quantitatively a degree of resistance between susceptible and resistant strains. For this test the RF for thiabendazole and levamisole was 14.3 and >32.5, respectively in H. contortus strains and 21.1 and 3.5 in strains of O. circumcincta. Egg hatch assay was also found to be sensitive and accurate in determining of resistance to benzimidazole. For measurement of levamisole resistance the egg hatch paralysis assay and larval paralysis assay were found to be able to distinguish between strains, but some disadvantages of these techniques make it unsuitable for field detection of levamisole resistance. Other in vitro assays as larval paralysis assay with physostigmine and larval micromotility assay were also investigated. Significant differences in paralysis of the larvae were observed using larval paralysis assay.

Animals↗

Age related occurrence of natural agglutinins in the erisilkworm, Philosamia ricini.

Natural agglutinins against human, guinea pig, and rat erythrocytes (RBC) and bacteria Bacillus thuringiensis were readily detected in hemolymph of final instar larvae with resulting titers of 0-9 (log2). Titers were independent of sex and season but varied conspicuously and reproducibly with age. Moreover, response of hemagglutinins to heating varied with age being heat-labile in younger larvae, yet totally resistant to heating for 30 min at 70 degrees C in older ones. Bacterial agglutinins were uniformly resistant at all larval ages. The study thus reveals that the amounts and physico-chemical properties of P. ricini agglutinins change with larval development. Therefore, larval age should be taken into close consideration before reporting on the occurrence and properties of agglutinins in insects.

Age Factors↗

Teratogenic and anti-metamorphic effects of bisphenol A on embryonic and larval Xenopus laevis.

Effects of bisphenol A (BPA) on embryonic and larval development were investigated. In Xenopus laevis blastulae treated with 2.5-3.0 x 10(-5) M BPA or with 10(-5) M 17beta estradiol (E2), malformation of the head region, scoliosis (curved vertebrate), and suppression of organogenesis were observed. In addition, 10(-5)-10(-4) M BPA blocked tri-iodothyronine (T3)-inducible resorption of the tail segments from premetamorphic (stage 52-54) larvae in vitro. When stage 52 tadpoles were immersed in 1.0-2.5 x 10(-5) M BPA, deceleration of both spontaneous and thyroxin (T4)-induced metamorphic changes occurred. Furthermore, BPA suppressed thyroid hormone receptor (TR) beta gene expression both in vivo and in vitro. Thus, we concluded that BPA at the concentrations examined affects both embryonic development and larval metamorphosis.

Animals↗

Cheating is not always punished: killer female plants and pollination by deceit in the dwarf palm Chamaerops humilis.

Because the interests of mutualists are not perfectly aligned, conflicts between partners often arise, rendering mutualism unstable by allowing the evolution of cheating. The dwarf palm Chamaerops humilis is engaged in a nursery pollination mutualism with a specific weevil Derelomus chamaeropsis. In exchange for pollen dispersal, dwarf palms provide pollinators with food, shelter and egg-laying sites, but pollinators can develop only within male inflorescences. Here we show that weevils lay eggs in female inflorescences but processes associated with fruit development prevent larval development. The cost imposed by developing larvae probably differs between male and female plants, explaining why only females defend their inflorescences. Female palms thus cheat their pollinating weevil, and pollinators are expected to 'punish' (avoid) them. We found no evidence for such punishment: weevils visit female plants and the duration of visits to male and female inflorescences does not differ. Thus mutualists do not always co-operate and cheating may not be necessarily punished.

Animals↗

Mass rearing the weevil Hylobius transversovittatus (Coleoptera: Curculionidae), biological control agent of Lythrum salicaria, on semiartificial diet.

Lythrum salicaria L., purple loosestrife, an invasive Eurasian perennial, is degrading wetlands across temperate North America. Chemical, physical, and mechanical methods failed to provide long-term control. Therefore, four host-specific insect species (two leaf feeders, a root feeder, and a flower feeder) were introduced as biological control agents. To increase the availability of adults of the root feeding weevil Hylobius transversovittatus Goeze for field releases, we developed a semiartificial diet. Suitability of different diet formulations (varying vitamin mixes, salt mixes, antimicrobials, water content, root content) and temperatures for larval development were evaluated. We also monitored how rearing on artificial diet and the number of larvae per container affected larval development time, larval survival, adult weight, and incidences of deformities. Rearing larvae on artificial diet reduced development time from 1-2 yr to 2-3 mo. Larval development was fastest and survival rates highest under constant temperatures of 25 degrees C. Hatch rate and larval survival decreased if eggs were surface sterilized. Using FABCO antimicrobials could not prevent fungal contamination; use of methyl paraben and sorbic acid was successful in suppressing fungal and bacterial growth throughout larval development time to <10%. The moisture content of the diet did not significantly affect larval survival, development, or adult weight. Decreasing the proportion of purple loosestrife roots in the diet reduced adult weight and the proportion of larvae completing development, and increased development time; no larvae were able to complete development in root-free diet. With an increase in the number of larvae per cup, survival rates were reduced. The experiments revealed a female biased sex ratio: females consistently developed faster and were heavier than males. Incidence of adult deformities was consistently below 5%. Increased availability of adults for field release as a result of mass production using the semiartificial diet will accelerate the biocontrol program targeting purple loosestrife. We are able to produce several hundred weevils per week and have adapted the diet to rear other root-feeding weevils.

Animals↗

Neuronal development in larval polychaete Phyllodoce maculata (Phyllodocidae).

The existing view on neuronal development in polychaetes, as undergoing neurogenesis beginning in the rudiments of central ganglia and then extending peripherally, has been contrasted with the latest findings in molluscs, their sister trochozoan group, which show a peripheral to central mode of neurogenesis. The current study addresses this issue by examining early neuronal development in the polychaete Phyllodoce maculata using immunolabeling against acetylated alpha-tubulin, serotonin, and the FMRFamide. The first nervous cell was detected 20 hours before hatching, at the early trochophore stage. A solitary serotonergic neuron was located at the posterior-dorsal extreme of the larva and issued anterior projecting fibers, which outline the future ventral nerve cords and prototroch nerve. Two more serotonergic dorsal peripheral cells and three peripheral FMRFamidergic cells appeared soon thereafter. The fibers of these early cells formed a scaffolding, which prefigured the future adult nervous system (cerebral ganglion, ventral cords, prototroch and esophageal nerve rings) in prehatched trochophores. Shortly before hatching, the larval nervous system developed, including the apical organ, meridianal nerves in the episphere, and posttrochal nerves that innervate the feeding apparatus. After hatching, the rudiments of the adult nervous system started to develop along the paths already established by the earliest peripheral neurons. Thus, the general strategy of neurogenesis in a representative polychaete trochophore appears to resemble that of molluscs. The first neuronal cells to appear are peripheral in origin, located near the posterior margins of the embryo. Their similar anatomical appearance suggests that they share a similar functional role in trochophore development and behavior.

Animals↗

Apical sensory neurones mediate developmental retardation induced by conspecific environmental stimuli in freshwater pulmonate snails.

Freshwater pond snails Helisoma trivolvis and Lymnaea stagnalis undergo larval development and metamorphosis inside egg capsules. We report that their development is permanently under slight tonic inhibitory influence of the anterior sensory monoaminergic neurones, which are the remnants of the apical sensory organ. Conspecific juvenile snails, when reared under conditions of starvation and crowding, release chemical signals that are detected by these neurones in encapsulated larvae and reversibly suppress larval development, thus providing a link between environmental signals and developmental regulation. Induced retardation starts from the trochophore stage and results in up to twofold prolongation of the larval lifespan. Upon stimulation with the signal, the neurones increase synthesis and release of monoamines [serotonin (5-HT) in Helisoma and dopamine in Lymnaea] that inhibit larval development acting via ergometrine-sensitive internal receptors. Thus, the novel regulatory mechanism in larval development of molluscs is suggested and compared with the phenomenon of dauer larvae formation in the nematode Caenorhabditis elegans.

5-Hydroxytryptophan↗

Circadian regulation of a daily rhythm of release of prothoracicotropic hormone from the brain retrocerebral complex of Rhodnius prolixus (hemiptera) during larval-adult development.

A daily rhythm of release of prothoracicotropic hormone (PTTH) has been reported throughout most of larval-adult development in Rhodnius prolixus. PTTH released by explanted brain-retrocerebral complexes was quantified using an in vitro bioassay in which the PTTH released into the incubation medium was assayed by its ability to stimulate ecdysteroid synthesis in arrhythmic prothoracic glands (PGs). The present article employs this assay to reveal that the daily rhythm of PTTH release is under circadian control. The rhythm free-runs in both continuous darkness (DD) and continuous light (LL) with a period length close to 24 hr, which is temperature compensated. The rhythm appears to damp out more rapidly in LL than in DD. It is argued that the circadian clock regulating PTTH release is in the brain and is entrained by extra-retinal photoreception. It is suggested that this "PTTH clock" is coupled in vivo to the clock previously described in the PGs that regulates rhythmic ecdysteroid synthesis by the circadian rhythm of release of PTTH. This coupling appears to be tight, since the rhythm of PTTH release retains close synchrony with the rhythm of ecdysteroid synthesis under both DD and LL. It is concluded that these two coupled clocks comprise a multioscillator system that drives the rhythms in ecdysteroid synthesis and the hemolymph ecdysteroid titer and consequently imposes temporal order on ecdysteroid-dependent developmental events.

Animals↗

Evaluation of the effects of selamectin against adult and immature stages of fleas (Ctenocephalides felis felis) on dogs and cats.

The adulticidal, ovicidal, and larvicidal effects of selamectin against flea (Ctenocephalides felis felis) infestations on dogs and cats were evaluated in a series of seven controlled and masked studies (three in cats, four in dogs). Animals were randomly allocated to treatment with either selamectin at a minimum dosage of 6mgkg(-1) in the commercial formulation or one of two negative-controls (0.9% NaCl solution or the vehicle from the commercial formulation). Treatments were administered topically in a single spot on the skin at the base of the neck in front of the scapulae. Speed of kill, measured by flea comb counts at 12h intervals during the 48h immediately following a single treatment on day 0, was evaluated in two studies. One study was in dogs and the other in cats, and each animal was infested with approximately 100 unfed viable adult fleas prior to treatment. Reductions in geometric mean flea counts for selamectin compared with saline were >98% between 24 and 36h after treatment in dogs, and between 12 and 24h after treatment in cats (P< or =0.0006). Efficacy in reducing flea egg hatch and larval development was evaluated in four studies, in which dogs and cats were treated once on day 0 and then repeatedly infested with approximately 600 fleas. Flea eggs were collected approximately for 72h after each infestation, on days 3, 7, 14, 21, and 30, counted, and cultured to determine their hatchability and subsequent larval development. Compared with the vehicle, selamectin was highly effective in reducing flea egg hatch (>92% in cats) and larval development (> or =95% for dogs and cats), and emergence of adults (97.8-100% for dogs, 85.6-100% for cats) for 30 days. Effects of exposure to hair coat debris were investigated in a study with dogs treated once on day 0 and repeatedly infested with 100 adult fleas. Debris (dander, flea faeces, hair, scales) was collected on days 1, 7, 14, 21, and 30 and added to normal flea eggs or larvae for incubation. Compared with debris from vehicle-treated dogs, debris from selamectin-treated dogs was highly effective in preventing egg hatch (>96%), in killing larvae (>98%) and in preventing larval development to adults (>99%) (P</=0.0033). Selamectin was shown to be highly effective in the treatment and control of flea infestations (C. felis) on dogs and cats. The adulticidal, ovicidal, and larvicidal effects of selamectin will be important in interrupting the flea life cycle by preventing the introduction and establishment of new flea infestations in a household environment.

Administration, Topical↗