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Efficiency of feeding Duddingtonia flagrans chlamydospores to control nematode parasites of first-season grazing goats in France.

A field trial, conducted over two consecutive years, was aimed at assessing the efficacy of the administration of spores of the nematophagous fungus Duddingtonia flagrans to young goats for the control of nematode parasite infections on a French commercial dairy goat flock. For both years, the first-year grazing kids were divided into two similarly managed groups (fungus and control groups): in 2003 a daily dose rate of 5 x 10(5) spores/kg body weight was given to the fungus-group animals, while in 2004 a daily dose rate of 10(6) spores/kg body weight was used; the other half of the kids, acting as control, did not receive the spores. Parameters measured every 3 weeks included nematode egg excretion, larval development in faecal cultures and pasture larval counts. Additionally, at the beginning, the middle and the end of each grazing season, the goats were weighed and blood samples for pepsinogen determination were collected. In 2003, similar results were recorded for all the measured parameters in the control and fungus groups. In contrast, in 2004, the kids receiving the spores showed lower faecal egg counts and pepsinogen levels at the end of the season and higher growth rate compared to kids of the control group.

Animals↗

Mytilin B and MGD2, two antimicrobial peptides of marine mussels: gene structure and expression analysis.

Previous research has shown that mytilins and MGDs are two types of 4-kDa, cysteine-rich, cationic antimicrobial peptides, which are abundant in hemocytes of the mussels, Mytilus galloprovincialis and M. edulis. The expression of the genes encoding these peptides has been analyzed in the hemocytes of animals subjected to various stress factors, as well as during larval development. Variations in gene expression in adult mussels have been tested under conditions of physical stress, bacterial challenge and heat shock. The results suggest that in adult mussels, the MGD2 gene may be over-expressed with physical and temperature stress, but that reduced expression occurs with bacterial challenge. Gene expression during development has been analyzed using different larval and post-larval stages, ranging from 4-day-old veliger larvae to 32-day-old post-larvae. The results show that the expression of both mytilin B and MGD2 is developmentally regulated, but neither gene is expressed in mussels until after larval settlement and metamorphosis. Finally, the genes encoding two isoforms of these peptides have been cloned and sequenced, revealing that both genes contain four exons and three introns.

Amino Acid Sequence↗

Control of developmental timing in Caenorhabditis elegans.

Studies of the nematode Caenorhabditis elegans have identified genetic and molecular mechanisms controlling temporal patterns of developmental events. Mutations in genes of the C. elegans heterochronic pathway cause altered temporal patterns of larval development, in which cells at certain larval stages execute cell division patterns or differentiation programs normally specific for other stages. The products of the heterochronic genes include transcriptional and translational regulators and two different cases of novel small translational regulatory RNAs. Other genes of the pathway encode evolutionarily conserved proteins, including a homolog of the Drosophila Period circadian timing regulator, and a member of the nuclear receptor family of proteins. These regulators interact with each other to elaborate stage-specific regulatory switches and act through downstream effectors to control the timing of cell-type-specific developmental events.

Animals↗

The E75 ecdysone-inducible gene responsible for the 75B early puff in Drosophila encodes two new members of the steroid receptor superfamily.

A pulse of the steroid hormone ecdysone at the end of Drosophila larval development triggers coordinate changes in both larval and imaginal tissues that result in metamorphosis to the adult fly. In larval salivary glands, this pulse activates a genetic regulatory hierarchy manifested by the induction of two kinds of transcription puffs in the polytene chromosomes: a small set of "early" puffs representing a primary response to the hormone, and a complex set of "late" puffs whose delayed appearance is dependent on proteins synthesized during the primary response. We isolated a 50-kb ecdysone-inducible gene, E75, that occupies the early puff locus at 75B. E75 contains two overlapping transcription units. The E75 A unit is a coextensive with the E75 gene and contains six exons: two 5'-proximal exons, A0 and A1, which are specific to this unit, and exons 2-5, which are common to both units. The E75 B unit is 20 kb long and contains five exons, a 5'-terminal exon, B1, located within the second intron of E75 A, and the common exons 2-5. Large open reading frames start within the first exon of each unit and continue into the last exon and therefore encode two different proteins. Both proteins exhibit sequence similarity to the conserved DNA-binding and hormone-binding domains of proteins in the steroid receptor superfamily. The two putative zinc fingers that characterize the DNA-binding domain are encoded by exon A1 and exon 2, so that the E75 A protein contains both fingers, whereas the E75 B protein contains only the second. Both proteins contain the same putative hormone-binding domain encoded by exon 4.

Amino Acid Sequence↗

Effect of larval population density on dynamics of growth in Chrysomyia albiceps (Wiedemann). I. Effect of feeding behaviour and growth rate of larvae.

The feeding behaviour of uncrowded and crowded larvae differed as soon as feeding began. In case of high density populations, larvae tend to form dense clusters and this gregariousness is responsible for much of the injury and resulting predation. Larval growth rate had the same pattern up to 200 larvae per 20 grams meat, then this pattern greatly changed with the increase of densities. Larval survivorship remained high at densities up to 160 larvae per 20 gm meat, then decreased rapidly as larval density increased. The duration of larval development was extended and synchronization of larval pupal ecdysis was affected. Percentage of pupation and pupal weight were greatly decreased at high densities.

Animals↗

Sampling, seasonal abundance, and mermithid parasitism of larval Coquillettidia perturbans in south-central Michigan.

Seasonal abundance of immature Coquillettidia perturbans was studied at 3 sites in south-central Michigan in 1987 and 1988. A modified boat bilge pump and funnel separators proved to be good sampling and sorting devices. Pupae were found in late May to early July, first and second instars were most abundant in July and August, while third and fourth instars were present year-round. Overall, larvae were most numerous in August and September, indicating that Cq. perturbans has a single generation per year in Michigan, with midsummer to fall as the main period of larval development, and with third and fourth larval instars as overwintering stages. Parasitism of Cq. perturbans larvae by a previously undiscovered mermithid nematode was documented at 2 sites in late summer.

Animals↗

A developmental study of serotonin-immunoreactive neurons in the larval central nervous system of the spider crab Hyas araneus (Decapoda, Brachyura).

Larval development in crabs is characterized by a striking double metamorphosis in the course of which the animals change from a pelagic to a benthic life style. The larval central nervous system has to provide an adequate behavioural repertoire during this transition. Thus, processes of neuronal reorganization and refinement of the early larval nervous system could be expected to occur in the metamorphosing animal. In order to follow identified sets of neurons throughout metamorphosis, whole mount preparations of the brain and ventral nerve cord of laboratory reared spider crab larvae (Hyas araneus) were labelled with an antibody against the neurotransmitter serotonin. The system of serotonin-immunoreactive cell bodies, fibres and neuropils is well-developed in newly hatched larvae. Most immunoreactive structures are located in the protocerebrum, with fewer in the suboesophaegeal ganglia, while the thoracic and abdominal ganglia initially comprise only a small number of serotonergic neurons and fibres. However, there are significant alterations in the staining pattern through larval development, some of which are correlated to metamorphic events. Accordingly, new serotonin-immunoreactive cells are added to the early larval set and the system of immunoreactive fibres is refined. These results are compared to the serotonergic innervation in other decapod crustaceans.

Aging↗

Cellular expression patterns of acetylcholinesterase activity during grasshopper development.

We examined the expression of acetylcholinesterase (AChE) in the nervous system and epidermal body structures during embryonic and larval development of two grasshopper species: Locusta migratoria and Schistocerca americana. Histochemical labelling was blocked by the enzyme inhibitors eserine and BW284c51, but not by iso-OMPA, showing that the staining reflected true AChE activity. The majority of staining was localized on the cell surface but granular intracellular staining was also visible in many cell bodies. In both species, the cellular expression of AChE followed a similar but complex spatiotemporal staining pattern. Initially, mainly epidermal tissue structures were stained in the various body appendages (stages 25%-30%). Labelling subsequently appeared in outgrowing neurons of the central nervous system (CNS) and in the nerves innervating the limbs and dorsal body wall (stages 30%-40%). The latter staining originated in motoneurons of the ventral nerve cord. In a third phase (after 45%), the somata of certain identified mechanosensory neurons started to express AChE activity, presumably reflecting cholinergic differentiation. Staining was also found in repo-positive glial cells of the CNS, longitudinal glia of connectives, glia of the stomatogastric nervous system and glial cells ensheathing peripheral nerves. Glial cells remained AChE-positive during larval to adult development, whereas motoneurons lost their AChE expression. The expression pattern in non-neuronal cells and glutamatergic motoneurons and the developmental appearance of AChE prior to synaptogenesis in the CNS suggest non-cholinergic functions of AChE during grasshopper embryogenesis.

Acetylcholinesterase↗

Effects of parasitism by Asobara tabida (Hymenoptera: Braconidae) on the development, survival and activity of Drosophila melanogaster larvae.

The impact of parasitism by Asobara tabida on Drosophila melanogaster larval development, survival features and larval activity has been investigated using two strains of the parasitoid. The successful parasitism rate of the A1 strain was four times greater than that of the WOPV strain. Both strains induced equivalent mortality rates but hosts parasitized by A1 predominantly died as pupae. The time necessary for the host pupariation and emergence, and the larval weight at 72, 96 and 120 h post-parasitization were measured. Parasitized larvae exhibited longer periods of development and lower weights than controls, especially when parasitized by A1. These results suggest that hosts underwent physiological costs varying with respect to the outcome of the parasitic relationship. Of the parasitoid factors possibly responsible for these costs, we examined venoms for their impact on host mortality. Artificial injections of WOPV venoms induced higher mortality rates than did A1 venoms. Venoms were also found responsible for the induction of a transient paralysis, naturally occuring after parasitization. Again, the strongest effect was observed after parasitization by WOPV or injections of its venoms. This study gives new insights into the intriguing features of A. tabida and constitutes the first report of the paralysing properties of the venoms.

Journal Article↗

Different pathways of DNA synthesis are operative in the fat body of Spodoptera litura (insecta) during larval and pupal development.

In Spodoptera litura, the DNA content of the fat body increases during the last larval instar. Radiolabelling studies reveal high degree of H3-thymidine incorporation in the fat cell DNA, during last larval stage of development. At prepupal stage the DNA synthesised during larval development is degraded, at least partially and the resultant nucleotides are most likely used as building blocks for the fat cell DNA synthesis during pupal development and pupal-adult metamorphosis, because very little H3 -thymidine injected into pupae is incorporated in DNA, though in fact, significant amount of DNA is synthesised during this period.

Animals↗

The impact of selective oviposition and infection with Plagiorchis elegans on Aedes aegypti pre-imago population dynamics at optimal food availability.

Progressive changes in the attraction of waters harboring pre-imago populations of Aedes aegypti exposed to different levels of the entomopathogenic digenean Plagiorchis elegans to ovipositing conspecific females were assessed under conditions of optimal food availability. The impact of ovipositional preference and parasitic infection on population structure and development was investigated. Probabilities that larvae progress from one stage to the next or die within 24 h were calculated for all life stages. Exposure to P. elegans cercariae did not significantly affect the attractiveness of larval-holding waters. Ovipositional preference increased significantly with growing bio-mass of the larval population, with the event of pupation and, in some cases, with late instar mortality. Exposure to various levels of the parasite significantly increased mortality of all instars, but most of the damage caused by the parasite occurred in the form of increased pupal mortality and decreased adult emergence. Exposure to the parasite significantly reduced the number of adults produced yet did not impair larval development. Thus, larval recruitment into environments containing P. elegans remains high, the structure of larval populations remains relatively normal, but few adults are produced.

Analysis of Variance↗

DHR3 is required for the prepupal-pupal transition and differentiation of adult structures during Drosophila metamorphosis.

Pulses of the steroid hormone ecdysone activate genetic regulatory hierarchies that coordinate the developmental changes associated with Drosophila metamorphosis. A high-titer ecdysone pulse at the end of larval development triggers puparium formation and induces expression of the DHR3 orphan nuclear receptor. Here we use both a heat-inducible DHR3 rescue construct and clonal analysis to define DHR3 functions during metamorphosis. Clonal analysis reveals requirements for DHR3 in the development of adult bristles, wings, and cuticle, and no apparent function in eye or leg development. DHR3 mutants rescued to the third larval instar also reveal essential functions during the onset of metamorphosis, leading to lethality during prepupal and early pupal stages. The phenotypes associated with these lethal phases are consistent with the effects of DHR3 mutations on ecdysone-regulated gene expression. Although DHR3 has been shown to be sufficient for early gene repression at puparium formation, it is not necessary for this response, indicating that other negative regulators may contribute to this pathway. In contrast, DHR3 is required for maximal expression of the midprepupal regulatory genes, EcR, E74B, and betaFTZ-1. Reductions in EcR and betaFTZ-F1 expression, in turn, lead to submaximal early gene induction in response to the prepupal ecdysone pulse and corresponding defects in adult head eversion and salivary gland cell death. These studies demonstrate that DHR3 is an essential regulator of the betaFTZ-F1 midprepupal competence factor, providing a functional link between the late larval and prepupal responses to ecdysone. Induction of DHR3 in early prepupae ensures that responses to the prepupal ecdysone pulse will be distinct from responses to the late larval pulse and thus that the animal progresses in an appropriate manner through the early stages of metamorphosis.

Animals↗

Azadirachtin potentiates the action of ecdysteroid agonist RH-2485 in Spodoptera littoralis.

Edysteroid agonist RH-2485 induces an immediate and fatal molt in Spodoptera littoralis when added to the diet of the 2nd and 4th instar larvae at 1 ppm, and to that of the 6th instar larvae at 0.001 ppm concentration. Ten times lower doses fed to the larvae continuously allow an apparently normal larval development that is terminated by a supernumerary larval molt. The other effects of RH-2485 include death during metamorphosis and impaired fertility of emerged adults. The number of progeny is reduced even with low RH-2485 doses that do not interfere with moltings; e.g., insects fed 0.0001 ppm since the 2nd, 4th, and 6th instar produce 72%, 62%, and 22%, respectively, less progeny than the controls. Feeding larvae with 10-1000 ppm Suneem oil (containing about 0.1-10 ppm azadirachtin) causes, in a stage- and dose-dependent manner, a cessation or reduction of feeding, delay of molts, death of larvae and pupae, and sterility of emerged adults; with 10 ppm Suneem oil, the number of progeny is reduced by 20-32%. Presence of Suneem oil in the diet does not influence the potential of RH-2485 to induce a prompt molt, but it increases ten times the potency to elicit a supernumerary larval molt. Certain combinations of RH-2485 with Suneem oil provoke up to 3 extra larval molts. Lethal developmental derangements and sterility are more frequent, and the response of larvae of different age is more uniform, when Suneem oil and RH-2485 are combined than when each of these agents is administered alone.

Journal Article↗

Structure and expression of daf-12: a nuclear hormone receptor with three isoforms that are involved in development and aging in Caenorhabditis elegans.

During Caenorhabditis elegans early larval development environmental conditions promote a cascade of signaling molecules to direct growth to the reproductive adult or to arrest development as a dauer larva. Two parallel chemosensory signal transduction pathways, one of which is transforming growth factor (TGF)-beta-like, converge on the daf-12 gene to regulate dauer formation. A third insulin-like signaling pathway interacts with the daf-12 pathway to regulate both dauer formation and adult longevity. To further understand the role of daf-12 in these processes, we have molecularly characterized this gene. We establish rescue of the mutant dauer defective phenotype with a genomic clone. We show that three transcripts of different lengths, due to differential splicing, are made from the daf-12 gene. The deduced protein isoforms are similar to both DNA- and ligand-binding domains of nuclear hormone receptors. The three daf-12 transcripts are produced throughout development and expression increases during the preparation for and execution of dauer formation. Analysis of various daf mutant strains suggests that the isoform ratios of daf-12 steady-state mRNA are not changed by reduction of function in the TGF-beta and insulin signaling components of the dauer pathway. The daf-12 promoter directs expression of GFP in the pharynx. daf-12 is a C. elegans nuclear hormone receptor with multiple isoforms, is expressed throughout development in distinct cells, and functions under a variety of environmental conditions.

Aging↗

Seasonal availability of nematode larvae on pastures grazed by cattle in New South Wales.

Trends in the numbers of infective menatode larvae on pasture plots contaminated by cattle at different seasons of the year were defined in 3 different climatic regions. The main nematodes were Ostertagia ostertagi, Trichostrongylus spp, Haemonchus spp and Cooperia spp. On the North Coast of New South Wales with a sub-tropical climate, the numbers of infective larvae of all 4 nematodes rose rapidly to peak levels soon after each seasonal period of contamination began, then fell quickly within a few months. On the Central Coast of New South Wales, the trends were similar to those on the North Coast, except that the larvae persisted on the pasture for a much longer time. On the Northern Tablelands of New South Wales, where temperatures were much colder than on the coast, larval development was slower and major peaks of larval availability did not occur until early spring. These different seasonal trends in each region were considered to be related to the climatic differences between the regions. On pastures which were contaminated continuously, larval numbers reached maximum levels in mid-winter on the Central Coast and in early spring on the Northern Tablelands. It was concluded that the majority of these larvae were derived from the contamination of pastures in autumn and winter. Subsequently in summer, a rapid dying out of larvae was observed in all the regions, probably due to the effect of hotter weather. The studies suggest that a reduction in the contamination of pasture with nematode eggs in autumn and winter could result in pastures carrying fewer larvae and thus form the basis of effective worm control programs for cattle.

Animals↗

Increased synaptic microtubules and altered synapse development in Drosophila sec8 mutants.

BACKGROUND: Sec8 is highly expressed in mammalian nervous systems and has been proposed to play a role in several aspects of neural development and function, including neurite outgrowth, calcium-dependent neurotransmitter secretion, trafficking of ionotropic glutamate receptors and regulation of neuronal microtubule assembly. However, these models have never been tested in vivo. Nervous system development and function have not been described after mutation of sec8 in any organism. RESULTS: We identified lethal sec8 mutants in an unbiased forward genetic screen for mutations causing defects in development of glutamatergic Drosophila neuromuscular junctions (NMJs). The Drosophila NMJ is genetically malleable and accessible throughout development to electrophysiology and immunocytochemistry, making it ideal for examination of the sec8 mutant synaptic phenotype. We developed antibodies to Drosophila Sec8 and showed that Sec8 is abundant at the NMJ. In our sec8 null mutants, in which the sec8 gene is specifically deleted, Sec8 immunoreactivity at the NMJ is eliminated but immunoblots reveal substantial maternal contribution in the rest of the animal. Contrary to the hypothesis that Sec8 is required for neurite outgrowth or synaptic terminal growth, immunocytochemical examination revealed that sec8 mutant NMJs developed more branches and presynaptic terminals during larval development, compared to controls. Synaptic electrophysiology showed no evidence that Sec8 is required for basal neurotransmission, though glutamate receptor trafficking was mildly disrupted in sec8 mutants. The most dramatic NMJ phenotype in sec8 mutants was an increase in synaptic microtubule density, which was approximately doubled compared to controls. CONCLUSION: Sec8 is abundant in the Drosophila NMJ. Sec8 is required in vivo for regulation of synaptic microtubule formation, and (probably secondarily) regulation of synaptic growth and glutamate receptor trafficking. We did not find any evidence that Sec8 is required for basal neurotransmission.

Animals↗

PAR-3 is required for epithelial cell polarity in the distal spermatheca of C. elegans.

PAR-3 is localized asymmetrically in epithelial cells in a variety of animals from Caenorhabditis elegans to mammals. Although C. elegans PAR-3 is known to act in early blastomeres to polarize the embryo, a role for PAR-3 in epithelial cells of C. elegans has not been established. Using RNA interference to deplete PAR-3 in developing larvae, we discovered a requirement for PAR-3 in spermathecal development. Spermathecal precursor cells are born during larval development and differentiate into an epithelium that forms a tube for the storage of sperm. Eggs must enter the spermatheca to complete ovulation. PAR-3-depleted worms exhibit defects in ovulation. Consistent with this phenotype, PAR-3 is transiently expressed and localized asymmetrically in the developing somatic gonad, including the spermathecal precursor cells of L4 larvae. We found that the defect in ovulation can be partially suppressed by a mutation in IPP-5, an inositol polyphosphate 5-phosphatase, indicating that one effect of PAR-3 depletion is disruption of signaling between oocyte and spermatheca. Microscopy revealed that the distribution of AJM-1, an apical junction marker, and apical microfilaments are severely affected in the distal spermatheca of PAR-3-depleted worms. We propose that PAR-3 activity is required for the proper polarization of spermathecal cells and that defective ovulation results from defective distal spermathecal development.

Animals↗

The influence of environmental P(O(2)) on hemoglobin oxygen saturation in developing zebrafish Danio rerio.

Several studies suggest that during early larval development of lower vertebrates convective blood flow is not essential to supply oxygen to the tissues, but information about the oxygenation status of larvae during the time of cutaneous respiration is still missing. If convective oxygen transport contributes to the oxygen supply to tissues, venous blood in the central circulatory system should be partly deoxygenated, and hyperoxia should increase the oxygen saturation of the hemoglobin. To analyze the changes in hemoglobin oxygen saturation induced by hyperoxic incubation, zebrafish larvae were incubated in a tiny chamber between polytetrafluoroethylene membranes (Teflon), so that the oxygen supply could be rapidly modified. Hemoglobin oxygen saturation was measured in vivo by combining video imaging techniques with a spectrophotometrical analysis of hemoglobin light absorption at specific wavelengths for maximal absorption of oxygenated and deoxygenated blood (413 nm and 431 nm, respectively) under normoxic conditions and after a 10 min period of hyperoxia (P(O(2))=100 kPa), assuming that at a P(O(2)) of 100 kPa the hemoglobin is fully saturated. The results demonstrated that red blood cell oxygenation of zebrafish larvae at 4 days post fertilization (d.p.f.), 5 d.p.f. and 12 d.p.f. could be increased by hyperoxia. The data suggest that at the time of yolk sac degradation (i.e. 4 d.p.f. and 5 d.p.f.), when the total surface area of the animal is reduced, bulk diffusion of oxygen may not be sufficient to prevent a partial deoxygenation of the hemoglobin. The decrease in hemoglobin oxygenation observed at 12 d.p.f. confirms earlier studies indicating that at 12-14 d.p.f., convective oxygen transport becomes necessary to ensure oxygen supply to the growing tissues.

Analysis of Variance↗