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The effect of p-aminobenzoic acid and folic acid on the development of infective larvae of Brugia malayi in Aedes aegypti.

Adult Aedes aegypti mosquitoes, infected with the subperiodic Brugia malayi, were found to enhance the development of the filarial parasites to the infective stage when they were exposed to a cotton pad soaked in 10% sucrose solution containing p-aminobenzoic acid (PABA) in 0.001, 0.005, 0.01, 0.05 and 0.1% concentrations. Similarly, larval development increased when the mosquitoes were fed with folic acid at 0.001, 0.01 and 0.1% concentrations. This stimulation was more when PABA or folic acid was given prior to the infected blood meal through the developmental period of the larvae. The data thus suggest that PABA and folic acid are nutrients for the development of B. malayi-microfilariae to the infective stage in A. aegypti.

4-Aminobenzoic Acid↗

How does blood cell concentration modulate cardiovascular parameters in developing zebrafish (Danio rerio)?

Blood flow and shear forces are considered to be important parameters possibly stimulating angiogenesis or cardiovascular remodeling. The main objective of this study was to test the hypothesis that a significant reduction in shear forces as a consequence of a significant isovolemic anemia induced by microsurgical techniques during early larval development of the zebrafish might induce a compensatory stimulation of erythropoiesis and/or induce a modification of cardiac activity or even the formation of the heart and may influence the shaping of the vascular bed. Blood from 2 day old zebrafish larvae was withdrawn and replaced by zebrafish Ringer's solution, so that the blood cell concentration was reduced by at least 75%. At 5 days post fertilization (dpf) a partial recovery in blood cell concentration was observed and reached a value of 814.55+/-85.42 cells/nL, while in control animals blood cell concentration amounted to 1856.00+/-131.59 cells/nL. At 7 dpf the value of blood cell concentration was 1023.89+/-95.75 cells/nL versus 1701.54+/-146.03 cells/nL in control animals. Compared to control animals, heart rate and cardiac output were significantly reduced in anemic animals and alterations in the formation of the vascular bed were also observed. A significant decrease in the end-diastolic volume suggested that ventricular volume was reduced. Thus, within a few days zebrafish larvae were nearly able to compensate for an isovolemic anemia by an enhanced erythropoiesis. However, several changes in cardiovascular system indicated that phenotypic plasticity is established even at an early developmental stage.

Anemia↗

Cellular mechanisms governing synaptic development in Drosophila melanogaster.

The neuromuscular connections of Drosophila are ideally suited for studying synaptic function and development. Hypotheses about cell recognition can be tested in a simple array of pre- and postsynaptic elements. Drosophila muscle fibers are multiply innervated by individually identifiable motoneurons. The neurons express several synaptic cotransmitters, including glutamate, proctolin, and octopamine, and are specialized by their synaptic morphology, neurotransmitters, and connectivity. During larval development the initial motoneuron endings grow extensively over the surface of the muscle fibers, and differentiate synaptic boutons of characteristic morphology. While considerable growth occurs postembryonically, the initial wiring of motoneurons to muscle fibers is accomplished during mid-to-late embryogenesis (stages 15-17). Efferent growth cones sample multiple muscle fibers with rapidly moving filopodia. Upon reaching their target muscle fibers, the growth cones rapidly differentiate into synaptic contacts whose morphology prefigures that of the larval junction. Mismatch experiments show that growth cones recognize specific muscle fibers, and can do so when the surrounding musculature is radically altered. However, when denied their normal targets, motoneurons can establish functional synapses on alternate muscle fibers. Blocking synaptic activity with either injected toxins or ion channel mutants does not derange synaptogenesis, but may influence the number of motor ending processes. The molecular mechanisms governing cellular recognition during synaptogenesis remain to be identified. However, several cell surface glycoproteins known to mediate cellular adhesion events in vitro are expressed by the developing synapses. Furthermore, enhancer detector lines have identified genes with expression restricted to small subsets of muscle fibers and/or motoneurons during the period of synaptogenesis. These observations suggest that in Drosophila a mechanism of target chemoaffinity may be involved in the genesis of stereotypic synaptic wiring.

Animals↗

Developmental ontogeny of prolactin and its receptor in fish.

Prolactin (PRL) is a member of a family of structurally similar proteins which includes growth hormone (GH) and somatolactin (SL) in teleost fish. The genes encoding these proteins are expressed principally in the pituitary gland and sequence analysis reveals they share considerable similarity. GH, PRL, and SL bring about their physiological action by binding to specific receptors localised in the membrane of cells in target tissue. The PRL receptor (PRLR) and GH receptor (GHR) have been identified in a number of teleosts but the SL receptor remains to be characterised. On hormone binding, receptors dimerise, and signal transduction occurs via the JAK/STAT signalling pathway. The principal action of PRL in fish is freshwater osmoregulation, although it has also been implicated in reproduction, behaviour, growth, and immunoregulation. The role of PRL in early development and metamorphosis is well established, respectively, in mammals and amphibians, although its role in fish is not so well known. Studies have shown that PRL mRNA and protein are restricted to the developing pituitary gland in fish embryos and larvae. PRLR mRNA and protein is also present in fish embryos and has a widespread tissue distribution in larvae. The levels of PRLR and PRL mRNA vary throughout embryonic and early larval development. The potential role of PRL in fish embryos and larvae is considered in relation to their physiological status.

Aging↗

[Morphology and development of the cysticercoids of Paricterotaenia porosa (Cestoda: Dilepididae)].

Studies of the development of Paricterotaenia porosa in its intermediate host Chironomus sp. were carried out. At 20 degrees C the development of the larva is completed in 24 days. The primary cavity is formed in 9 days, the separation of the cercomere anlage--in 11 to 12 days and the invagination of the larva--in 19 days. The larval development of P. porosa is characterized by its passing in the cavity of the external cyst, by the complete separation of anlages of the body of the larva and cercomere at the metamere stage and by the intensive growth of the cercomere after the invagination of the larva. The analysis of the cercomere structure suggests its protective function.

Animals↗

In vivo and in vitro activity of venom from the endoparasitic wasp Pimpla turionellae (L.) (Hymenoptera: Ichneumonidae).

The biological activity of venom from Pimpla turionellae L. (Hymenoptera: Ichneumonidae) was examined in vivo toward larvae and pupae of Galleriae mellonella L. (Lepidoptera: Pyralidae), and in vitro toward bacterial and fungal cultures, as well as cultured insect cells. Pupae of G. mellonella were far more susceptible to the venom than larvae. At low doses of venom [0.1 venom reservoir equivalents (VRE)], pupal abdominal mobility was inhibited within 30 min, and by 24 h, all pupae injected with venom concentrations >0.5 VRE were completely paralyzed. These same doses of venom resulted in an inhibition of adult emergence. Host larvae were far less sensitive to wasp venom as evidenced by all venom injected larvae remaining responsive to mechanical stimulation by 1 h post injection, even at concentrations equivalent to 1 venom reservoir. Eventually (>2 h at 25 degrees C), venom-injected larvae became immobile, then flaccid, and all died within 24 h post-injection. At lower concentrations of wasp venom, the onset of paralysis was delayed by comparison to that evoked by 1 VRE, and few host larvae were able to pupate. Development of host larvae to adult emergence was also reduced in a dose-dependent manner, with eclosion completely prevented at high concentrations (>0.5 VRE) of venom. Venom doses <0.5 VRE did not appear to induce paralysis or alter larval development. When venom was incubated with bacterial or fungal cultures, no antimicrobial activity was detected. However, wasp venom was found to be cytotoxic and cytolytic to cultured cells derived from the cabbage looper Trichoplusia ni Hubner (Lepidoptera: Noctuidae) and the yellow fever mosquito, Aedes aegypti (L.) (Diptera: Culcidae). Though both cell types displayed similar susceptibility in terms of LC50s, the lepidopteran cells responded much more rapidly with regard to the onset of morphological changes and the timing of cell death. A possible mode of action for the venom is discussed.

Animals↗

The Drosophila dosage compensation complex binds to polytene chromosomes independently of developmental changes in transcription.

In Drosophila, the dosage compensation complex (DCC) mediates upregulation of transcription from the single male X chromosome. Despite coating the polytene male X, the DCC pattern looks discontinuous and probably reflects DCC dynamic associations with genes active at a given moment of development in a salivary gland. To test this hypothesis, we compared binding patterns of the DCC and of the elongating form of RNA polymerase II (PolIIo). We found that, unlike PolIIo, the DCC demonstrates a stable banded pattern throughout larval development and escapes binding to a subset of transcriptionally active areas, including developmental puffs. Moreover, these proteins are not completely colocalized at the electron microscopy level. These data combined imply that simple recognition of PolII machinery or of general features of active chromatin is either insufficient or not involved in DCC recruitment to its targets. We propose that DCC-mediated site-specific upregulation of transcription is not the fate of all active X-linked genes in males. Additionally, we found that DCC subunit MLE associates dynamically with developmental and heat-shock-induced puffs and, surprisingly, with those developing within DCC-devoid regions of the male X, thus resembling the PolIIo pattern. These data imply that, independently of other MSL proteins, the RNA-helicase MLE might participate in general transcriptional regulation or RNA processing.

Animals↗

[Age specificity of the effect of 5-bromdeoxyuridine on the differentiation and puff spectrum of the salivary gland chromosomes of Drosophila virilis Sturt. I. Relationship between the viability of individuals and induction of new puffs and the period of action of 5-bromdeoxyuridine].

5-Bromodeoxyuridine (BUdR) influences the differentiation of the imaginal discs, larval viability and the chromosome puffing throughout the larval development. The differentiation of imaginal discs is more sensitive during the early 3rd instar. "cryptocephal" phenocopy appeared if the larvae were reared in the BUdR medium till the late 2nd instar. 2nd and 3rd instars are critical for the larval viability. During the period of the middle 3rd instar BUdR induced the complete spectrum of the newly forming specific puffs.

Age Factors↗

Digestion of delta-endotoxin by gut proteases may explain reduced sensitivity of advanced instar larvae of Spodoptera littoralis to CryIC.

The present study describes the correlation between gut protease activity of lepidopteran larvae of different instars, the inactivation of Bacillus thuringiensis delta-endotoxins in crystalline and noncrystalline forms, and the reduced susceptibility of advanced larval instars of Spodoptera littoralis to the toxin. The original assembly of delta-endotoxins in a crystal structure is essential for causing efficient larval mortality. Denaturation and renaturation (D/R) of delta-endotoxin crystals increased the vulnerability of the toxin molecules to proteolysis, reduced their capability to kill neonate larvae of S. littoralis, but sustained most of their larval growth-inhibition activity. E. coli-produced CryIC delta-endotoxin applied as a fraction of inclusion bodies exerted a growth inhibition effect, similar to the molecules released from the crystals by denaturation and subsequent renaturation. Incubation of CryIC with gut juice of 1st or 2nd instar larvae, left part of the CryIC toxin intact, while the toxin was completely degraded when incubated with gut juice of 5th instar larvae. The degradation rate was consistent with the increase of protease specific activity of the gut juice during larval development. This increase in toxin degradation may account for the loss of sensitivity of 5th instar larvae to CryIC. Specific protease inhibitors such as PMSF and Leupeptin were shown to inhibit gut proteases activity in all instar larvae, while, 1,10 phenanthroline, TLCK and TPCK were effective only in young instar larvae. The differential effect of protease inhibitors on proteases obtained from different larval instars indicated that gut juice protease profiles change with larval age. The observed quantitative and qualitative differences in degradation of delta-endotoxin by larval gut proteases that occur during larval maturation may account for the difference in susceptibility to the delta-endotoxin. This finding should be taken into consideration when designing strategies for the development of transgenic crops expressing delta-endotoxins as potent insecticidal proteins.

Animals↗

Systematic identification of oscillatory gene expression in single cell types.

Many biological cycles are driven by oscillatory gene expression coordinated across cell types. For example, larval development in Caenorhabditis elegans involves coordinated cyclic changes in cell division, behavior, and growth, the latter requiring production of a structured extracellular matrix called the cuticle. Here, we combine single-cell RNA sequencing and novel computational approaches to identify oscillatory gene expression in individual cell types. We find that many cell types exhibit looping structures in PCA and UMAP space that correspond to transcriptional oscillations at each larval stage. Oscillatory gene expression is found in all cuticle-producing cell types, including glia, but not detected in neurons or muscle. We develop rigorous statistical approaches for de novo identification of oscillatory genes and cell types, yielding >5,000 genes. While many oscillatory genes relate to cuticle production, each cell type expresses largely distinct genes, suggesting that cuticle production is a patchwork of cell-type-specific programs. Finally, we derive a potential set of regulatory transcription factors that can explain coordinated oscillatory gene expression and find that shared upstream factors likely control gene timing across cell types. Together, our results suggest that shared regulators control cell-type-specific oscillatory gene expression, including in previously overlooked cell types such as glia.

Journal Article↗

The effects of dimethylated and alkylated polycyclic aromatic hydrocarbons on the embryonic development of the Japanese medaka.

The Japanese medaka (Oryzias latipes) early-life stage assay was used to investigate the effects of a number of commercially available dimethylated polycyclic aromatic hydrocarbons (PAHs) (3,6-dimethylphenanthrene, 7,12-dimethylbenz[a]anthracene, and 4,6-dimethyldibenzothiophene) and their unsubstituted congeners, dimethylated and unsubstituted tertiary mixtures, and a complex environmental mixture (with elevated C2-substituted dibenzothiophene) on embryo larval development. Unsubstituted PAHs showed trends of increased blue sac disease (BSD) relative to dimethylated PAHs, although the severity of BSD induction varied. Results demonstrated that the dibenzothiophene congeners were the strongest inducers of BSD of the commercial PAHs tested. These compounds reduced the hatching success of embryonic medaka, an effect that was enhanced in the mixture. The base neutral extract significantly increased the frequency and severity of BSD abnormalities, while significantly reducing larval hatch length. Based on these results, a sublethal maximum allowable toxicant concentration (MATC) of 13.91 microg PAHs/L was calculated.

Animals↗

Effects of pond water, sediment and sediment extract samples from New Hampshire, USA on early Xenopus development and metamorphosis: comparison to native species.

In an effort to assess potential ecological hazards to amphibian species in selected regions within New Hampshire, the traditional Frog Embryo Teratogenesis Assay-Xenopus (FETAX), a 14-/21 day tail resorption thyroid disruption assay and >30 day limb development tests were conducted with representative surface water and sediment samples. Two separate sets of samples collected from five sites were evaluated. The primary objectives of the study were to determine if samples were capable of inducing early embryo-larval maldevelopment, to determine if maldevelopment included limb defects, to determine if thyroxine co-administration altered the rates of limb malformation and to evaluate the impact of the samples on growth rates, developmental progress and metamorphic climax. Results from these studies suggested that pond water and sediment extract samples, but not whole sediment samples, from B2, FW, LP and W ponds were capable of inducing abnormal early embryo-larval development. In addition, water samples from B2 and W ponds induced significant abnormal hindlimb development. Some abnormal forelimb development was noted in the tail resorption studies, but not to the same extent as the hindlimbs. Each of the water samples induced appreciable developmental delay, including the paired reference site B1, which could be reversed by the addition of exogenous thyroxine.

Animals↗

Metabolic rate variation in Glossina pallidipes (Diptera: Glossinidae): gender, ageing and repeatability.

Despite the importance of metabolic rate in determining flight time of tsetse and in mediating the influence of abiotic variables on life history parameters (and hence abundance and distribution), metabolic rate measurements and their repeatability have not been widely assessed in these flies. We investigate age-related changes in standard metabolic rate (SMR) and its repeatability, using flow-through respirometry, for a variety of feeding, gender and pregnancy classes during early adult development in laboratory-reared individuals of the tsetse fly, Glossina pallidipes. Standard metabolic rate (144-635 microW) was generally within 22% of previous estimates, though lower than the values found using closed system respirometry. There was no significant difference between the genders, but metabolic rate increased consistently with age, probably owing to flight muscle development. Repeatability of metabolic rate was generally high (r=0.6-.09), but not in younger teneral adults and pregnant females (r approximately equal to 0.05-0.4). In these individuals, low repeatability values are a consequence of muscle or in utero larval development. Tsetse and other flies generally have a much higher metabolic rate, for a given size, than do other insect species investigated to date.

Aging↗

Low larval vector survival explains unstable malaria in the western Kenya highlands.

Several highland areas in eastern Africa have recently suffered from serious malaria epidemics. Some models predict that, in the short term, these areas will experience more epidemics as a result of global warming. However, the various processes underlying these changes are poorly understood. We therefore investigated malaria prevalence, malaria vector densities and malaria vector survival in a highland area in western Kenya, ranging from approximately 1,550-1,650 m altitude. Although only five adult malaria vectors were collected during 180 light traps and 180 resting collections over a 23-month study period, malaria was prevalent among school children (average parasite prevalence: 10%). During an extensive survey of potential larval habitats, we identified only seven habitats containing Anopheles gambiae Giles s.l. larvae. Their limited number and low larval densities suggested that their contribution to the adult vector population was small. Experiments on adult and larval survival showed that at this altitude, adult mosquitoes survived inside local houses, but that larval development was severely retarded: only 2 of 500 A. gambiae s.l. larvae developed to the pupal stage, whereas all other larvae died prior to pupation. At present, high vector densities are unlikely because of unfavourable abiotic conditions in the area. However, temporary favourable conditions, such as during El Niño years, may increase larval vector survival and may lead to malaria epidemics.

Altitude↗

The effects of ozone-exposed sugar maple seedlings on the biological performance and the feeding preference of the forest tent caterpillar (Malacosoma disstria Hbn.).

The effects of exposure of sugar maple (Acer saccharum Marsh.) to ozone on the entire larval stage of a native insect have not been previously investigated. This study reports the effects of sugar maple seedlings exposed to different ozone concentrations on the relative performance and the feeding preference of the forest tent caterpillar (Malacosoma disstria Hbn.). Three-year-old seedlings were set in nine open-top field chambers in the spring of 1992 and 1993. Three ozone concentrations were generated: charcoal-filtered ambient air (0x), ambient air (1x) and three times ambient air (3x). In 1992, female and male larval development time did not differ among ozone treatments. In 1993, female larvae reared on 3x developed faster than those on 0x and 1x, while male larvae were not affected. Ozone treatments did not influence pupal weights except for males in 1993 where pupae reared on 0x were heavier than 1x but did not differ from 3x. Larval and pupal survival rates were not affected by ozone in either year. Finally, 4th and 5th instar larvae showed a significant feeding preference for 3x foliage in 1993 but not in 1992. The response of the forest tent caterpillar to ozone exposed seedlings varied between years and could be more sensitive to annual climatic variations than ozone.

Journal Article↗

Molecular characterization and developmental expression of a retinoid- and fatty acid-binding glycoprotein from Drosophila. A putative lipophorin.

A detailed understanding of the mechanism of lipid transport in insects has been hampered by the inability to identify the proapolipophorin gene that encodes apolipophorins I and II, the principal protein components of lipophorin, the lipid transport vehicle. Here we provide the first molecular description of the Drosophila gene encoding a retinoid- and fatty acid-binding glycoprotein (RFABG) and present evidence that it is a member of the proapolipophorin gene family. The gene, localized to the chromosome 4 (102 F region), encodes a 3351-amino acid protein that could serve as the precursor for the approximately 70-kDa and >200-kDa polypeptides associated with RFABG. The N-terminal sequence of the approximately 70-kDa polypeptide and that predicted for the >200-kDa polypeptide showed high sequence similarity to blowfly apolipophorin II and apolipophorin I, respectively. The RFABG precursor contains a signal peptide and exhibits a multidomain mosaic protein structure, which is typical of extracellular proteins. It has structural domains similar to lipid-binding proteins, namely vitellogenins and apolipoprotein B. The protein also contains a domain similar to the D domain of von Willebrand factor and mucin. The gene is expressed in the Drosophila embryo during development in cells that make up the amnioserosa and fat bodies. Immunolocalizations using specific antibodies against RFABG reveal that the protein is initially dispersed through the embryonic amnioserosa sac and latter concentrated at skeletal muscle-epidermis apodemeal contact junctions during larval development. This novel gene may play an important role in the transport of lipids, including retinoids and fatty acids, in insects.

Amino Acid Sequence↗

Susceptibility of mulesing wounds to oviposition and strike development by the Australian sheep blowfly Lucilia cuprina.

The age and/or physical condition of mulesing wounds had a significant effect on both the oviposition response of L. cuprina and the ability of the wound to support a strike. Up to 48 h after mulesing, untreated wounds elicited a strong oviposition response in contrast with chemically treated wounds, although subsequent larval development was negligible. Seven to 9-day-old wounds, however, regardless of the wound treatment, were highly attractive oviposition sites, which subsequently developed into strikes; the D3 formulation of Defiance*S, however, significantly depressed strike development and shows promise as a mules wound treatment. A marked improvement in the wound healing 14 to 16 d after mulesing coincided with a significant decrease in oviposition. Only those sheep whose wound scabs remained broken, exposing pus and raw tissue, attracted oviposition; subsequent development of the eggs into strikes was negligible.

Animals↗

Oestrogens have no hormonal effect on the development and reproduction of the harpacticoid copepod Nitocra spinipes.

In recent years, reports have described endocrine-disruptive effects of environmental oestrogens in fish, but little is known about similar effects in crustaceans. The objective of the present study was therefore to examine whether the oestrogens 17-beta-oestradiol, 17-alpha-ethynylestradiol and diethylstilbestrol (DES), could affect mortality, larval development rate, fecundity and sex ratio in the sexually reproducing harpacticoid copepod Nitocra spinipes. Newly released nauplii (<24-h old) were exposed to 1/1,000, 1/100 and 1/10 (nominal concentrations) of each oestrogen's 96 h-LC50 value for < or = 18 days at 22 +/- 1 degrees C. The percentage of gravid females and the number of developed copepodites were both reduced at 0.03 mg l(-1) DES, although the latter response was not significant. None of the other two oestrogens induced any measurable effects. Since the only observed significant response appeared at a DES concentration no more than 10 times below the 96 h-LC50 value, there is no evidence of endocrine-disruptive activity in N. spinipes exposed to oestrogens.

Animals↗