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PubMed2000

Onchocerca-Simulium complexes in Venezuela: can human onchocerciasis spread outside its present endemic areas?

The compatibility between sympatric and allopatric combinations of Onchocerca volvulus-anthropophilic species of Simulium was studied in the north-eastern focus of human onchocerciasis as well as in a densely populated locality of the Amazonas State in Venezuela. The objectives were to test the conjecture that local adaptation exists between the parasite and its vectors (the Onchocerca-Simulium complex hypothesis), and assess the possibility of the infection spreading from its present distributional range. For the homologous combination, O. volvulus-S. metallicum cytospecies E in Anzoátegui State (north-eastern focus), parasite yield was 45% in contrast to 1% for the heterologous, southern parasite-S. metallicum infection. This was significantly lower than the parasite yield (4-10%) expected after allowing for the effect of density-dependent limitation of infective larval output described in this paper for S. metallicum. The population of S. exiguum s.l. from southern Venezuela allowed no larval development beyond the L1 stage of either northern or southern parasites. Mechanisms for such refractoriness probably operate at the level of the thoracic muscles, not affecting microfilarial uptake or migration out of the bloodmeal. The parasite yield of southern O. volvulus in S. oyapockense s.l. flies biting man at Puerto Ayacucho (Amazonas) was about 1%, in agreement with the figures recorded for highly compatible sympatric combinations such as O. volvulus-S. ochraceum s.l. in Guatemala. No infective larval development of the northern parasite was observed in southern S. oyapockense. These results, together with considerations of typical worm burdens in the human host, presence/absence of armed cibaria in the simuliids, parasite-induced vector mortality, and fly biting rates, suggest a lower potential for onchocerciasis to spread between the northern and southern endemic areas of Venezuela than that between Amazonian hyperendemic locations and settlements outside this focus with high densities of S. oyapockense s.l.

Adult↗
PubMed2005

The ecdysone-induced DHR4 orphan nuclear receptor coordinates growth and maturation in Drosophila.

A critical determinant of insect body size is the time at which the larva stops feeding and initiates wandering in preparation for metamorphosis. No genes have been identified that regulate growth by contributing to this key developmental decision to terminate feeding. We show here that mutations in the DHR4 orphan nuclear receptor result in larvae that precociously leave the food to form premature prepupae, resulting in abbreviated larval development that translates directly into smaller and lighter animals. In addition, we show that DHR4 plays a central role in the genetic cascades triggered by the steroid hormone ecdysone at the onset of metamorphosis, acting as both a repressor of the early ecdysone-induced regulatory genes and an inducer of the betaFTZ-F1 midprepupal competence factor. We propose that DHR4 coordinates growth and maturation in Drosophila by mediating endocrine responses to the attainment of critical weight during larval development.

Animals↗
PubMed1998

Evidence of multiple mechanisms of avermectin resistance in haemonchus contortus--comparison of selection protocols.

Three isolates of Haemonchus contortus selected for avermectin resistance in sheep were compared in three in vitro pharmacological tests previously shown to discriminate between field isolates of H. contortus resistant and susceptible to the avermectins. Two isolates, F7-A and IVC, were selected for avermectin resistance in the laboratory from a reference susceptible isolate using suboptimal doses of ivermectin (LD95) for 7 and 16 generations, respectively. In these isolates avermectin resistance was not associated with a decreased sensitivity to avermectin inhibition of larval development or L3 motility but was associated with an increased sensitivity to paraherquamide. The third isolate, Warren, was derived from an overwhelmingly avermectin-susceptible, mixed species field isolate in a single generation by propagating the small number of survivors of a 0.2 mg/kg ivermectin treatment (i.e. 10 x LD95). This isolate, like previously characterised avermectin-resistant H. contortus isolates derived from the field in South Africa and Australia, showed a markedly reduced sensitivity to avermectin inhibition of larval development and L3 motility, as well as an increased sensitivity to paraherquamide. These results suggest that avermectin resistance can manifest itself in different ways and that the two selection protocols used to generate the F7-A, IVC and Warren isolates have resulted in the selection of different resistance phenotypes.

Animals↗
PubMed1998

Developmental changes in dopamine levels in larvae of the fly Chymomyza costata: comparison between wild-type and mutant-nondiapause strains.

Dopamine and two related catecholamines, L-3,4-dihydroxyphenylalanine (DOPA) and N-acetyl dopamine (NADA), were analyzed in whole body and tissue samples taken throughout late larval development of the drosophilid fly Chymomyza costata, which enters facultative diapause as a mature 3rd instar larva in response to short photophase. Wild-type (W) and mutant-nondiapause (M) strains reared under diapause inducing (d) and preventing (nd) photophases were compared. Developmental changes in the whole body of dopamine levels showed some general features irrespective of fly strain and rearing conditions: sharp major peaks during moult from second to third instar larva and during pupariation; lower minor peaks around the middle of both 2nd and 3rd instars. Significant differences between the strains and conditions were also found: dopamine levels were lower throughout the 2nd instar and during the 2nd to 3rd instar moult of mutant strain larvae (M/d) as compared to wild-type larvae (W/nd and W/d); while the late 2nd and late 3rd instar larvae destined to diapause (W/d) maintained relatively high dopamine concentrations, their counterparts destined to continuous development (W/nd and M/d) significantly decreased dopamine levels prior to the 2nd to 3rd instar moult or pupariation. Possible relationship between the dopamine levels and diapause induction/onset in C. costata larvae is discussed. Integument contained more than 90% of the dopamine found in the whole body. The gut and central nervous tissues showed relatively low pools of dopamine, only trace amounts were detected in haemolymph and no dopamine was found in fat body. DOPA levels were low and stable throughout larval development of both W and M strains and under both conditions. NADA levels peaked during second halves of 2nd and 3rd instars of both strains, then dropped to trace levels and were elevated again during 2nd to 3rd instar moult as well as in tanned prepupae. No elevation of NADA levels was recorded in 3rd instar W/d larvae which entered diapause.

Journal Article↗
PubMed1997

Proximo-distal specification in the wing disc of Drosophila by the nubbin gene.

Mutations in the nubbin (nub) gene have a phenotype consisting of a severe wing size reduction and pattern alterations, such as transformations of distal elements into proximal ones. nub expression is restricted to the wing pouch cells in wing discs since early larval development. These effects are also observed in genetic mosaics where cell proliferation is reduced in all wing blade regions autonomously, and transformation into proximal elements is observed in distal clones. Clones located in the proximal region of the wing blade cause in addition nonautonomous reduction of the whole wing. Cell lineage experiments in a nub mutant background show that clones respect neither the anterior-posterior nor the dorsal-ventral boundary but that the selector genes have been correctly expressed since early larval development. The phenotypes of nub el and nub dpp genetic combinations are synergistic and the overexpression of dpp in clones in nub wings does not result in overproliferation of the surrounding wild-type cells. We discuss the role of nub in the wing's proximo-distal axis and in the formation of compartment boundaries.

Animals↗
PubMed2001

Effects of a mustard trypsin inhibitor expressed in different plants on three lepidopteran pests.

The effects of mustard trypsin inhibitor MTI-2 expressed at different levels in transgenic tobacco, arabidopsis and oilseed rape lines have been evaluated against three different lepidopteran insect pests. 1. Plutella xylostella (L.) larvae were the most sensitive to the ingestion of MTI-2. The inhibitor expressed at high levels in arabidopsis plants caused rapid and complete mortality. High mortality and significantly delayed larval development were also detectable in oilseed rape expressing MTI-2 at lower levels. 2. Mamestra brassicae (L.) larvae were sensitive only at high MTI-2 expression level, as obtained in transgenic tobacco and arabidopsis, whereas no effects were observed for larvae fed on plants showing relatively low expression levels such as those of oilseed rape lines. 3. Feeding bioassays with Spodoptera littoralis (Boisduval) larvae were carried out using the same oilseed rape lines, showing that at these low expression levels no mortality was observed although a delay in larval development did occur. The levels of insect gut proteolytic activities of the larvae still alive at the end of a 7 day feeding bioassay were usually higher than in the controls, but no new proteinases were expressed in any case. The combined results described in this paper demonstrate altogether the relevance of a case-by-case analysis [target insects and proteinase inhibitor (PI) level of expression in planta] in a PI-based strategy for plant protection.

Animals↗
PubMed2001

The Drosophila genes disconnected and disco-related are redundant with respect to larval head development and accumulation of mRNAs from deformed target genes.

HOM-C/hox genes specify body pattern by encoding regionally expressed transcription factors that activate the appropriate target genes necessary for differentiation of each body region. The current model of target gene activation suggests that interactions with cofactors influence DNA-binding ability and target gene activation by the HOM-C/hox proteins. Currently, little is known about the specifics of this process because few target genes and fewer cofactors have been identified. We undertook a deficiency screen in Drosophila melanogaster in an attempt to identify loci potentially encoding cofactors for the protein encoded by the HOM-C gene Deformed (Dfd). We identified a region of the X chromosome that, when absent, leads to loss of specific larval mouthpart structures producing a phenotype similar to that observed in Dfd mutants. The phenotype is correlated with reduced accumulation of mRNAs from Dfd target genes, though there appears to be no effect on Dfd protein accumulation. We show that these defects are due to the loss of two functionally redundant, neighboring genes encoding zinc finger transcription factors, disconnected and a gene we call disco-related. We discuss the role of these genes during differentiation of the gnathal segments and, in light of other recent findings, propose that regionally expressed zinc finger proteins may play a central role with the HOM-C proteins in establishing body pattern.

Amino Acid Sequence↗
PubMed1990

Development of larval Echinococcus granulosus confronted with chicken heart tissue in vitro.

Vesiculated protoscoleces (VP) were produced by culturing freshly collected protoscoleces from Echinococcus granulosus horse liver hydatids in RPMI 1640 monophasic medium at 37 degrees C for 18 days. Half of the VP were used as such, the other half used after killing them by freeze-thawing. Nine-day-old chicken heart fragments (CHF) were cultured in MEM at 37 degrees C for 72 h. Subsequently, CHF were put together with live and dead VP, respectively, for up to 53 days, on a semisolid medium consisting of agar, Ringer's and MEM. Time-dependent histological observations revealed that dead VP were surrounded by CHF cells. Dead VP tissue was eventually internalized and disintegrated in about 1 week. Live VP penetrated into the CHF tissue and further developed into small hydatid cysts, located within the boundaries of the experimental 'host' tissue. The amorphous-looking contact region PAP-stained positively only with anti-E. granulosus serum and not with anti-CHF serum; it was considered identical to the normal laminated layer. The invasion of VP in CHF tissue proved to be different from a tumour or a bacterial invasion: it was concluded that the confrontation of VP and CHF had resulted in an 'in vitro cohabitation' rather than in an 'in vitro infection'.

Animals↗
PubMedDate not supplied

Toxicity of surficial sediments from Sydney Harbour and vicinity, Australia.

The toxicological responses of three species to 103 surficial saltwater sediment samples from Sydney Harbour, and coastal lakes and estuaries on the south-east coast of New South Wales, Australia, were tested in a battery of four to six laboratory toxicity tests. This is the first large-scale toxicological study of sediments in Australia, the objective of which is to assess the protective and predictive abilities of North American biological effects-based sediment quality guidelines, recently adopted in Australia. Amphipods were exposed to whole sediments in survival and reburial tests, sea urchin fertilisation and larval development tests were conducted on porewaters, and bacterial bio-luminescence (Microtox) tests were conducted on organic solvent extracts and porewaters. Local indigenous species were used for the amphipod and sea urchin tests (Corophium sp. and Heliocidaris tuberculata, respectively). A wide range of responses, from <25 to 100% of negative controls were observed in all tests. Mean control-adjusted responses ranged from 46 to 96% for all tests. The percentages of highly toxic samples ranged from 11 to 83% in the various tests. The order of test sensitivity was: amphipod survival < Microtox test of porewaters < amphipod reburial < sea urchin larval development < sea urchin fertilisation < Microtox test of solvent extracts. Concordance between toxicity tests in classifying samples as highly toxic or not, ranged from 47 to 79%, indicating some similarities between test results, but not complete equivalence. Combined toxicity test results showed that the incidence of highly toxic responses occurring in the majority of tests (75-100% of tests) was low (5% of samples), but a large percentage of samples had highly toxic results in at least one test (76% of samples). Toxicity was more pervasive in the Sydney region than in coastal lakes and estuaries south of Sydney. The current study demonstrated the utility of indigenous invertebrate species and the Microtox bacterium in a sediment toxicity test battery for Australian saltwater sediments.

Animals↗
PubMed1992

Ontogeny of substance P-, CGRP-, and VIP-containing nerve fibers in the amphibian carotid labyrinth of the bullfrog, Rana catesbeiana. An immunohistochemical study.

The ontogeny of substance P, CGRP (calcitonin gene-related peptide), and VIP (vasoactive intestinal polypeptide) containing nerve fibers in the carotid labyrinth of the bullfrog, Rana catesbeiana, was examined by the peroxidase-antiperoxidase method. The time of appearance of these three peptides was different for each. First, CGRP fibers appeared in the wall of the carotid arch and external carotid arteries, and in a thin septum between these two arteries at an early stage of larval development (stage III). At stage V, substance P immunoreactive fibers appeared, and VIP fibers were detected at the early metamorphic stage (stage XXII). Up to the completion of metamorphosis, the number of these fibers remained low. From 1 to 5 weeks after metamorphosis, substance P, CGRP, and VIP fibers increased in number to varying degrees. By 8 weeks after metamorphosis, the distribution and abundance of these fibers closely resembled those of the adults. Some CGRP and VIP immunoreactive glomus cells were found at the stages immediately before and after the completion of metamorphosis. These findings suggest that substance P, CGRP, and VIP fibers during larval development and metamorphosis may be nonfunctional, and start to participate in vascular regulation only after metamorphosis. The transient CGRP and VIP in some glomus cells may be important for the development of the labyrinth, or may take part in vascular regulation through the close apposition of the glomus and smooth muscle cells (g-s connection).

Animals↗
PubMed1998

Immunological studies on the settlement-inducing protein complex (SIPC) of the barnacle Balanus amphitrite and its possible involvement in larva-larva interactions.

Immunological investigation has revealed that a settlement-inducing protein complex (SIPC), which induces cypris settlement of the barnacle Balanus amphitrite, is synthesized during larval development and accumulates in the cypris larva. We previously purified the SIPC from adult B. amphitrite, which was active when bound to a substratum. The SIPC is a glycoprotein of high molecular mass, consisting of three major subunits of 76, 88 and 98 kDa with lentil lectin (LCA)-binding sugar chains. In the present study, we prepared antiserum against each LCA-binding subunit of SIPC, and performed immunoblot analyses. Immunoblotting of adult extracts showed that anti-76-kDa antibody reacted only with the 76-kDa protein, whereas anti-88-kDa and anti-98-kDa antibodies reacted with both the 88-kDa and the 98-kDa proteins. Immunoblotting of larval extracts indicated that reactivity of the 76-kDa protein to anti-76-kDa antiserum increased during larval development and cyprid extracts reacted strongly. Moreover, by using immunostaining we found that the SIPC was contained in 'footprints' of cyprids, which have been shown to act as a settlement-inducing pheromone, and is secreted onto the antennular attachment discs. The results suggest that the SIPC (or SIPC-like proteins) is involved in both adult-larva and larva-larva interactions during settlement of the barnacle B. amphitrite.

Animals↗
PubMed1994

Morphogenesis of catecholaminergic interneurons in the frog spinal cord.

During embryonic and larval development of the frog Xenopus laevis, a bilateral population of cerebrospinal fluid-contacting neurons matures in the ventral spinal cord. These cells are catecholaminergic and form a dispersed or nonrandom pattern of spacing within each of their spinal cord columns. In order to test the mechanisms underlying pattern formation of these neurons, it is first necessary to understand their normal morphogenesis. Morphogenetic changes were examined by using immunocytochemistry for tyrosine hydroxylase as a cell marker. Immunoreactivity in spinal cord cells was detected as early as 1.4 days (stage 28) of embryonic development. Subsequently, these cells underwent changes in shape, and rapid, regressive changes in cell size. The population emerges gradually during development, but the major characteristic of nonrandom spacing, their dispersion from other catecholaminergic cells, is apparent at early stages of differentiation. Increases in cell density occur over an extended period of time and can be divided into an initial phase of large, rapid changes and a subsequent plateau phase of gradual changes. The two longitudinal columns of catecholaminergic cells that are characteristic of older animals become apparent just before hatching, when density increases until cells on both sides of the midline are present in the same region. Although the dispersed pattern exists within each column, cross-correlation analysis shows that there is a random relationship between cells in opposite columns. During larval development, the catecholaminergic cell domain expands in both a rostral and caudal direction. The morphogenetic changes of the catecholaminergic cell population begin to show how the cells become partitioned within the floor plate region of the spinal cord.

Animals↗
PubMed2002

Functional genomics and sexual differentiation in amphibians.

In Xenopus laevis the basic mechanisms underlying sexual differentiation were investigated by determining time courses of sexual steroids and their corresponding receptors during complete larval development from egg to juveniles. Androgens as well as estradiol (E2) are derived from maternal origin and accumulate in hatching tadpoles. Sexual steroid contents decreased rapidly after hatching and rose again at the end of metamorphosis indicating endogenous production. In parallel the mRNA expression for corresponding androgen (AR) and estrogen receptors (ER) was measured by means of semiquantitative RT-PCR. Both receptor mRNAs increased dramatically just after hatching and decreased only moderately until end of metamorphosis. In female juveniles E2 and ER-mRNA levels were higher compared with males. Treatment by exogenous E2 elevated both, ER- and AR-mRNA, indicating stimulatory functions of E2 for gene expression of both receptors. Effects on sexual differentiation during larval development were achieved by treatment with E2 and the antiandrogen cyproterone acetate both causing feminization, the antiestrogen tamoxifen resulting in neutralization, and the androgens, methyltestosterone and dihydrotestosterone, but not testosterone, leading to masculinization. The data presented are in accordance with further recent findings and suggest a new hypothesis for functional genomics in sexual differentiation of amphibians.

Androgens↗
PubMedDate not supplied

Gene expression and larval evolution: changing roles of distal-less and orthodenticle in echinoderm larvae.

We describe the expression of the homeobox genes orthodenticle (Otx) and distal-less (Dlx) during the larval development of seven species representing three classes of echinoderms: Holothuroidea, Asteroidea, and Echinoidea. Several expression domains are conserved between species within a single class, including Dlx expression within the brachiolar arms of asteroid larvae and Otx expression within the ciliated bands of holothuroid larvae. Some expression domains are apparently conserved between classes, such as the expression of Dlx within the hydrocoel (left mesocoel) in all three classes. However, several substantial differences in expression domains among taxa were also evident for both genes. Some autapomorphic (unique derived) features of gene expression are phylogenetically associated with autapomorphic structures, such as Dlx expression within the invaginating rudiment of euechinoids. Other autapomorphic gene expression domains are associated with evolutionary shifts in life history from feeding to nonfeeding larval development, such as Otx expression within the ciliated bands of a nonfeeding holothuroid larva. Similar associations between evolutionary changes in morphology and life history mode with changes in regulatory gene expression have also been observed in arthropods, urochordates, and chordates. We predict that recruitment of regulatory genes to a new developmental role is commonly associated with evolutionary changes in morphology and may be particularly common in clades with complex life cycles and diversity of life history modes. Caution should be used when making generalizations about gene expression and function based on a single species, which may not accurately reflect developmental processes and life histories of the phyla to which it belongs.

Animals↗
PubMed1990

[Variation in the level of free amino acids during larva development of Penaeus japonicus (Crustacea, Decapoda, Penaeidae].

Variations of free amino-acids (FAA) content during the larval development of Penaeus japonicus have been studied. FAA content increases from 83 to 687 mumoles/g dry weight from egg to post-larval (20 days old: P20) development. There is a sharp increase between zoe and mysis stages, and another one between one day post-larvae (P1) and P20. After the mysis stage, free essential and non essential amino-acids evolve in a different way: the non essential FAA content, mainly glycine, increases continuously, whereas essential FAA content decreases (about 40%) from the mysis stage to metamorphosis in post-larvae (P1). FAA content evolution during larval development exhibits the same trend as the variation of the digestive enzyme activity.

Amino Acids↗
PubMed1986

Ontogeny of heart rate regulation in the bullfrog, Rana catesbeiana.

Heart rate (fH) at 20-23 degrees C was recorded in six different developmental stages of the bullfrog, Rana catesbeiana (n = 104, body mass 40 mg to 90 g), at rest after normoxic acclimation, during acute changes in O2 availability, and after brief but intense activity. The effects of cholinergic blockade and combined cholinergic and beta-adrenergic blockade on the response to this experimental protocol were also assessed. Mild tonic vagal inhibition of fH was evident during larval development but disappeared after metamorphosis. There was no tonic sympathetic stimulation of fH at rest in any developmental stage. Intense activity produced a tachycardia in all developmental stages but newly hatched larvae. In adults, tachycardia during activity resulted from beta-adrenergic stimulation but in larvae may have resulted from the direct effects of increased venous return stretching the cardiac pacemaker. Neither acute hypoxia or hyperoxia affected fH in any developmental stage, with the single exception of a severe depression of fH occurring at a PO2 of 30 Torr in newly hatched larvae. These results indicate that, although the heart of the newly hatched larvae is essentially devoid of regulation, cholinergic and beta-adrenergic mechanisms for reflex regulation of fH appear early in larval development. These mechanisms, although most fully expressed in the adult bullfrog, are essentially intact before metamorphosis of the larva.

Animals↗
PubMed2006

Influence of photon beam irradiation on Lymnaea natalensis snails infected with Fasciola gigantica.

Lymnaea natalensis is the intermediate host of Fasciola gigantica in Egypt. The effect of photon beam irradiation on the ability of the laboratory reared L. natalensis to support the larval development of F. gigantica has been studied. 120 snails were divided into two groups: The control infected non irradiated group (GI) and the experimental infected irradiated group (GII). The later group was subdivided into two subgroups: GIIa: snails irradiated before infection and GIIb: snails irradiated after infection. Photon beam irradiation had non significant effect on the survival rate between the all groups at the 30th day post infection. The life span, the number of infected snails and the length of the shedding period were significantly decreased in the two irradiated subgroups than the control group. The effect was more obvious on GIIb without significant difference. The number of metacercariae significantly decreased in the 2 irradiated subgroups than the control one. Also, it was significantly decreased in GIIb when compared with GIIa. So, photon beam irradiation has a great role on retarding larval development of F. gigantica inside the snail. This opens the way to a new strategy for fascioliasis control of in Egypt.

Animals↗
PubMed2000

Inheritance of avermectin resistance in Haemonchus contortus.

A larval development assay was used to compare the responses of the Chiswick Avermectin Resistant (CAVRS) isolate of Haemonchus contortus, an avermectin-susceptible isolate (VRSG) and their crosses to avermectins. The F(1) and F(2) generations of reciprocal crosses between CAVRS and VRSG were denoted as CAVRS malesxVRSG females=CXV, and VRSG malesxCAVRS females=VXC. The levels of avermectin resistance in the developing larvae of the F(1) of both CXV and VXC were indistinguishable from that in the avermectin-resistant parent, indicating that the resistance trait is completely dominant. Avermectin dose-response curves for the CXV F(1) did not show a 50% mortality rate at low concentrations, indicating that avermectin resistance is not sex-linked. This conclusion was confirmed when adult male worms of the F(1) of the CXV mating were found to have survived treatment of the host with 200microgkg(-1) ivermectin. This dose rate (200microgkg(-1) ivermectin) caused a 50% reduction in the number of adult males in the F(1) from both CXV and VXC crosses, but only a non-significant reduction in the number of adult females in the F(1). Dose-response curves obtained for the F(2) generations in the larval development assay indicated the presence of 25% of avermectin-susceptible individuals, suggesting that a single major gene largely controls the avermectin-resistance trait. This genetic analysis of avermectin resistance in an Australian H. contortus isolate indicates that the expression of the gene for avermectin resistance is an autosomal, complete dominant in the larvae; however, in adults its expression is sex-influenced, with males having a lower resistance to avermectin than females.

Animals↗