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Ontogeny of thyroid hormones, cortisol, hsp70 and hsp90 during silver sea bream larval development.

We studied the profiles of silver sea bream (Sparus sarba) thyroxine (T(4)), triiodothyronine (T(3)), cortisol and the heat shock protein (hsp) families hsp70 and hsp90 during larval development. Eggs from sexually mature female sea bream were fertilized and larvae were collected at incremental time intervals between 1-46 days post hatch (dph). Both T(4) and T(3) were detected in 1 dph larvae and it was found that both increased as development progressed with a distinct surge in amounts between 21-35 dph, a time associated with direct development of larvae to juveniles. Cortisol increased from 1 dph reaching a maximum and constant level from 35 dph onwards. Using RT-PCR coupled with radioisotope hybridization of immobilized cDNA we assessed the transcript levels of hsp70 and it was found that transcript remained unaltered between 1-14 dph before progressively increasing. Immunoblotting was used to study the larval concentrations of hsp70 and hsp90 and it was found that hsp70 was not significantly changed between 1-14 dph whereas hsp90 increased from 1 dph onwards. These findings suggest an important role for hsp90 in the corticosteroid receptor complex during silver sea bream larval development.

Animals↗

Interspecific competition among Aedes aegypti, Ae. albopictus, and Ae. triseriatus (Diptera: Culicidae): larval development in mixed cultures.

Interactions among the larvae of Aedes aegypti (L.), Ae. albopictus (Skuse), and Ae. triseriatus (Say) were studied in trispecific and bispecific mixed populations under laboratory and field conditions. Competitive stress (as evidenced by the average time to first, 50, and 75% pupation and the total pupation periods for mixed populations of each species in comparison with their single species controls) was more pronounced in mixed cultures reared in glass jars in the laboratory than in tires under field conditions. In the laboratory, the larval development of Ae. aegypti reared together with Ae. albopictus or Ae. triseriatus, or both, larvae was accelerated significantly. Conversely, the time to pupation for Ae. albopictus and Ae. triseriatus was delayed when reared with Ae. aegypti. However, the average wing length of female Ae. albopictus and Ae. triseriatus was greater in the mixed cultures than in single species cultures. These data indicated that the effect of intraspecific competition was greater than interspecific competition. Adequate food and higher temperature appeared to promote rapid development and higher survival of the immature stages of the three Aedes species in tires placed in the field. In general, Ae. triseriatus larvae required a longer period for larval development and had greater larval mortality than either Ae. aegypti or Ae. albopictus. In mixed populations of Ae, albopictus and Ae. aegypti in the jars and food-rich tires, the periods needed to attain first, 50, and 75% cumulative pupation were not significantly different than in single species controls. We conclude that no clear-cut displacement occurred in mixed experimental populations of Ae. aegypti and Ae. albopictus.

Aedes↗

Schistosoma rodhaini: intramolluscan larval development, migration and replication processes of daughter sporocysts.

The larval development of Schistosoma rodhaini in Biomphalaria glabrata presents two features of interest. The first concerns the migration of daughter sporocysts: only a fraction of the daughter sporocysts produced by the mother sporocyst migrate directly towards the digestive gland of the snail and become cercariogenous; the others remain in the anterior region of the snail where the majority transforms into small sporocystogenous sporocysts. The second point concerns the replication processes: only direct replication of daughter sporocysts was observed in sporocysts in a prehepatic localization. The demographic processes involved in the development of S. rodhaini are compared with other species of Schistosoma and interpreted as an adaptive character for better exploitation of the snail host.

Animals↗

Insulin-like growth factors I and II in the sole Solea senegalensis: cDNA cloning and quantitation of gene expression in tissues and during larval development.

Insulin-like growth factors I and II (IGF-I and IGF-II) play an important role as modulators of development, growth, and reproduction. This study aimed to isolate the IGF-I and IGF-II cDNAs and determine their temporal expression pattern in different organs and throughout larval development in Senegal sole. The rapid amplification of cDNA ends (RACE) was used to obtain both full-length IGFs sequences. A high sequence similarity with other teleosts sequences was observed. Domains B and A revealed as the most evolutionary conserved. Steady-state copy numbers of IGF-I and IGF-II were also quantified in different Senegal sole tissues by real-time PCR. IGF-I and IGF-II expressed ubiquitously with the highest mRNA levels in liver (88 x 10(6) molecules/microg total RNA) and gills (14.0 x 10(6) molecules/microg total RNA) respectively. IGF-II mRNA levels were higher than IGF-I in prehatching embryos and premetamorphic larvae with a significant drop before the commencement of eye migration in metamorphosis. The abundance of IGF-II transcripts correlated positively with the growth rate during larval development. The putative role of IGF-II on metamorphosis and larval growth is discussed.

Amino Acid Sequence↗

Identification and quantification of hydroxamic acids in maize seedling root tissue and impact on western corn rootworm (Coleoptera: Chrysomelidae) larval development.

Hydroxamic acid content was analyzed in the root tissue of four maize, Zea mays L., lines using high-performance liquid chromatography (HPLC) and related to western corn rootworm, Diabrotica virgifera virgifera LeConte, larval development and survivorship. Maize lines evaluated included Mp710 (PI 596627), MpSWCB-4, (PI 550498), Sc213 (PI 548792), and Dk580 (DeKalb commercial hybrid). Maize plants from each line were grown in test tubes containing a transparent agarose gel medium in a growth chamber. After 8 d of growth, root tissue of each line was harvested and hydroxamic acid content analyzed using HPLC. Three hydroxamic acids, 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), 6-methoxybenzoxazolinone (MBOA), and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), were identified in the maize roots tested. DIMBOA concentration was quantified and ranged from 246.37 +/- 70.53 micrograms to 91.84 +/- 49.82 micrograms DIMBOA per gram of root tissue. No significant difference was found among lines in D. v. virgifera larval development and survivorship.

Animals↗

Cuticle deposition and ecdysteroid titers during embryonic and larval development of the argasid tick Ornithodoros moubata (Murray, 1877, sensu Walton, 1962) (Ixodoidea:Argasidae).

Timing of embryonic and larval molts at the ultrastructural level and presence of ecdysteroids (ES) during embryonic and larval development of the argasid tick Ornithodoros moubata were studied. Embryonic "cuticles" A, B, and C were deposited 24-30 hr, 48-56 hr, and 6 days after oviposition, respectively. Deposition of the larval cuticulin layer started on Day 8 of embryonic development and procuticle deposition continued after hatching until apolysis of the larval cuticle 40 hr posthatch. Plaques of cuticulin formed on tips of microvilli 48-56 hr after hatching and procuticle was deposited until after ecdysis. Radioimmunoassay (RIA) was used to determine the ES titer in methanolic extracts of various ages of embryos and larvae. No peaks of RIA-positive material were detected during deposition of envelopes A, B, and C. However two peaks of ES were observed during embryonic development, one which coincided with the shortening of the germ band and a second which coincided with the deposition of the larval epicuticle on Day 8. During larval development, a peak of ES was observed on Day 3 (48-56 hr posthatch) and was correlated with nymphal epicuticle deposition. HPLC-RIA revealed that these last two peaks consisted mainly of 20-hydroxyecdysone together with a small quantity of ecdysone. Conjugated RIA positive material was present in freshly laid eggs and an augmentation of this esterase hydrolysable material was noted at the appearance of each ES peak. Thus the embryos did not appear to be hydrolyzing the maternal apolar conjugates to release ES during embryonic development; on the contrary, they appeared to be conjugating the newly synthesized hormones.

Animals↗

Elevated copper levels during larval development cause altered locomotor behavior in the adult carabid beetle Pterostichus cupreus L. (Coleoptera: Carbidae).

It is generally believed that copper causes changes in carabid communities indirectly by reducing food availability, because these animals are frequently found to have only slightly elevated metal contents even close to pollution sources. Using computer-centered video tracking, the locomotor behavior of adult Pterostichus cupreus carabid beetles was quantified after being raised on copper-contaminated food and soil during larval development. Copper was found to have an acute toxic effect measured in larval mortality, to cause a slight increase in the developmental period of males, but not to effect the emergence weights of adults of either sex. This toxic effect on the larvae was preserved through pupation to the surviving adults, which were normal in size and appearance, but displayed a dramatically depressed locomotor behavior. Copper analysis of these adults revealed that copper levels were either the same as or only slightly elevated in comparison with controls. The findings suggest that the altered locomotor behavior is associated with copper-induced internal structural damage during larval development and therefore expresses a prolonged or permanent effect. Such changes in locomotor behavior are likely to reduce the fitness of the animal under field conditions.

Animals↗

C. elegans knockouts in ubiquinone biosynthesis genes result in different phenotypes during larval development.

Ubiquinone is an essential molecule in aerobic organisms to achieve both, ATP synthesis and antioxidant defence. Mutants in genes responsible of ubiquinone biosynthesis lead to non-respiring petite yeast. In C. elegans, coq-7/clk-1 but not coq-3 mutants live longer than wild type showing a 'slowed' phenotype. In this paper we demonstrate that absence in ubiquinone in coq-1, coq-2 or coq-8 mutants lead to larval development arrest, slowed pharyngeal pumping, eventual paralysis and cell death. All these features emerge during larval development, whereas embryo development appeared similar to that of wild type individuals. Dietary coenzyme Q did not restore any of the alterations found in these coq mutants. These phenomena suggest that coenzyme Q mutants unable to synthesize this molecule develop a deleterious phenotype leading to lethality. On the contrary, phenotype of C. elegans coq-7/clk-1 mutants may be a unique phenotype than can not generalize to mutants in ubiquinone biosynthesis. This particular phenotype may not be based on the absence of endogenous coenzyme Q, but to the simultaneous presence of dietary coenzyme Q and the its biosynthesis intermediate demethoxy-coenzyme Q.

Aging↗

Partial substitution of di- and tripeptides for native proteins in sea bass diet improves Dicentrarchus labrax larval development.

To determine whether incorporation of peptides into diets can improve larval development, sea bass (Dicentrarchus labrax) larvae were fed for 21 d one of three isonitrogenous, isoenergetic semipurified diets in which enzymatic hydrolysate (75% di- and tripeptides) of fish meal proteins was substituted for 0, 20 or 40% of native fish meal proteins. Growth and survival were significantly greater (P < 0.05) in larvae fed peptide diets compared to those fed only native protein, with the best performance exhibited by those fed the 20% level of peptides. Chymotrypsin activity was much higher in groups fed peptide diets compared to that fed all native protein (P < 0.001), indicating a greater proteolytic capacity of the pancreas. At the intestinal level, activities of the brush border enzymes, aminopeptidase, maltase and gamma-glutamyl transpeptidase, increased with age while the cytosolic enzyme, leu-ala peptidase, decreased with age (P < 0.001). These changes in enzymatic activities correspond to the normal development of intestinal digestion. This development occurred earlier in the group fed 20% peptide-substituted diet than in the two other groups. The better larval performances observed in groups fed diets containing peptides can be related to the enhanced proteolytic capacity of the pancreas and the earlier development of intestinal digestion.

Aminopeptidases↗

Larval development test for detection of anthelmintic resistant nematodes.

The growth, using freshly cultured Escherichia coli with ampicillin or heat-treated lyophilised E coli as a food source, of the larvae of the mouse nematode Nematospiroides dubius and the infectivity of resulting third stage larvae were determined. Concentrations of E coli between 0.5 and 1 mg dry weight ml-1 permitted optimal larval development for both N dubius and Trichostrongylus colubriformis. Development of larvae of susceptible and cambendazole-resistant strains of Haemonchus contortus in thiabendazole solutions showed clear differences between the strains and the larval development test was more sensitive than the egg hatch test. The test also detected a levamisole resistant strain of H contortus, although the degree of resistance could not be adequately measured. It is concluded that the test can be run with any anthelmintic to which resistance is suspected.

Animals↗

Larval development and metamorphosis in Pleurobranchaea maculata, with a review of development in the notaspidea (Opisthobranchia).

Pleurobranchaea maculata is a carnivorous notaspidean that is common in New Zealand. This species produces small eggs (diameter 100 microm) and planktotrophic veligers that hatch in 8 d and are planktonic for 3 weeks before settling on biofilmed surfaces (14 degrees C). Larval development is known in detail for only two other notaspidean species, P. japonica and Berthellina citrina. In all three species of pleurobranchids, mantle and shell growth show striking differences from veligers of other opisthobranch taxa. In young veligers of pleurobranchids, the shell is overgrown by the mantle, new shell is added by cells other than those of the mantle fold, and an operculum does not form. Thus some "adult" traits (e.g., notum differentiation, mechanism of shell growth, lack of operculum) are expressed early in larval development. This suggests that apomorphies characteristic of adult pleurobranchids evolved through heterochrony, with expression in larvae of traits typical of adults of other clades. The protoconch is dissolved post-settlement and not cast off as occurs in other opisthobranch orders, indicating that shell loss is apomorphic. P. maculata veligers are atypical of opisthobranchs in having a field of highly folded cells on the lower velar surface, a mouth that is posterior to the metatroch, and a richly glandular, possibly chemodefensive mantle. These data indicate that notaspidean larvae are highly derived in terms of the novel traits and the timing of morphogenic events. Phylogenetic analysis must consider embryological origins before assuming homology, as morphological similarities (e.g., shell loss) may have developed through distinct mechanisms.

Animals↗

Genetic contribution to variation in larval development time, adult size, and longevity of starved adults of Anopheles gambiae.

The variation in mosquito life-history traits such as adult size has been studied with respect to environmental factors, but the genetic contribution to such variation has received almost no consideration. Using a full-sib design of F1s produced by wild caught Anopheles gambiae (M molecular form) females, we estimated broad-sense heritability of larval developmental time, adult size (based on dry weight and wing length), and longevity of starved adults. These traits were correlated (at the phenotypic level) with each other in females and males (|r(p)|>0.5, P<0.001). Longevity of starved adults increased with adult size, and both traits (adult longevity and size) decreased with longer larval development. Genetic correlations were lower (|r(g)|>0.45, P<0.05) but provided consistent evidence against a trade off between adult size and larval development time predicting that a mosquito can develop faster into a smaller adult or be a larger adult by a longer development. Estimates of heritability of the three traits were moderate to high (range: 0.05-0.48) and statistically significant (P<0.05), indicating substantial genetic contribution to the phenotypic variation in these traits. These results suggest that adaptive differences are likely to be found in these traits between A. gambiae populations.

Animals↗

Effect of condensed tannins on egg hatching and larval development of Trichostrongylus colubriformis in vitro.

The effects of condensed tannins extracted from seven forages on the viability of the eggs and first stage (L1) larvae of the sheep nematode Trichostrongylus colubriformis were evaluated in in vitro assays. The extracts of condensed tannins were obtained from Lotus pedunculatus (LP), Lotus corniculatus (LC), sulla (Hedysarum coronarium), sainfoin (Onobrychus viciifolia), Dorycnium pentaphylum (DP), Dorycnium rectum (DR) and dock (Rumex obtusifolius). Extracts containing 200 to 500 microg/ml reduced the proportion of eggs that hatched. The larval development assay was used to evaluate the effect of the extracts on the development of either eggs or L1 larvae to L3 infective larvae. Development was allowed to proceed for seven days by which time the larvae in control incubations had reached the infective L3 stage. Extracts containing 200 microg/ml from LP, DP, DR or dock prevented egg development, and only 11, 8 and 2 per cent of the eggs developed to L3 larvae with extracts from LC, sulla and sainfoin, respectively. When the concentration was 400 microg/ml no eggs developed to L3 larvae. The addition of the extracts after hatching also inhibited the development of L1 to L3 larvae; 200 microg/ml extracted from LP, LC, sulla, sainfoin, DP, DR and dock resulted in only 14, 18, 17, 15, 14, 16 and 4 per cent of L1 larvae developing to the L3 stage compared with 85 per cent for controls, and 400 microg/ml further reduced the development of L1 larvae. Statistical analyses showed that when the extracts were added before hatching they were significantly (P<0.001) more effective at inhibiting the larval development than when they were added after hatching. The condensed tannins from dock had the greatest inhibitory effect on egg development followed by the tannins from DR, sainfoin, DP, LP, sulla and LC.

Animals↗

Effects of tetrabromobisphenol A on larval development and thyroid hormone-regulated biomarkers of the amphibian Xenopus laevis.

Effects of tetrabromobisphenol A (TBBA) on thyroid hormone (TH)-dependent morphological development and on TH-regulated biomarkers were investigated using the amphibian Xenopus laevis. Tadpoles were exposed in the Xenopus metamorphosis assay to different TBBA concentrations ranging from 2.5 to 500 microg/L for 21 days. Only the highest TBBA concentration inhibited larval development while the biomarker for negative feedback mechanism, TH receptor beta-mRNA, was not affected. Short-term exposures to TBBA, triiodothyronine (T3), and a combination of TBBA and T3 were performed to examine expression of TH-upregulated biomarkers. TBBA alone increased slightly the expression of TH receptor beta- and basic region leucin zipper transcription factor b/Zip-mRNA but inhibited their T3-induced elevation in a dose-dependent manner. Despite the fact that TBBA seems to cause only mild effects during long-term treatment on larval development, short-term exposure revealed indirect evidence that TBBA can function as a TH antagonist.

Animals↗

Embryonic and larval development of the sonic motor nucleus in the oyster toadfish.

The sonic motor nucleus (SMN), a likely homologue of the hypoglossal nucleus, provides the final common pathway for sound production in the oyster toadfish (Opsanus tau). SMN neurons increase in size and number for 7-8 years postnatally, and the swimbladder-sonic muscle complex grows throughout life. This study describes the normal embryonic and larval development of the SMN from its initial differentiation on about day 19 through day 40, when the yolk sac is resorbed and the fish is free swimming. In contrast to the rapid development of CNS nuclei in mammals, the SMN gradually increased in maturity with more active growth at the beginning and end of the observation period and a relatively static period in the middle. Consistent with a hypoglossal homology, the SMN differentiated within the spinal cord, added cells rostrally, and eventually extended into the medulla. Immature neurons appeared to originate from precursor cells in the ventral portion of the ventricular zone of the central canal. Such cells were initially round with little cytoplasmic development and later added processes and Nissl substance. The number of neurons increased 10-fold from a median of 35 to 322 cells, and no evidence of cell death was observed. Soma area approximately doubled from 20.6 to 41.2 micron 2, and cell nucleus area followed a similar pattern. [3H]-thymidine autoradiography demonstrated that neurons were added continuously throughout the nucleus during embryonic and larval development.

Air Sacs↗

Larval development of Lynceus brachyurus (Crustacea, Branchiopoda, Laevicaudata): redescription of unusual crustacean nauplii, with special attention to the molt between last nauplius and first juvenile.

The larval development of "conchostracans" has received only scattered attention. Here I present the results of a study on the larval (naupliar) development and the metamorphosis of Lynceus brachyurus, a member of the bivalved branchiopod order the Laevicaudata. Lynceus brachyurus is the only species of the "Conchostraca" in Denmark. The phylogenetic position of the Laevicaudata has traditionally been a source of controversy, and this study does not solve the question completely. This work focuses on features potentially important for phylogeny. The general appearance of the larvae of L. brachyurus has been known for more than a century and a half, and some of its unique features include a large, larval dorsal shield; a huge, plate-like labrum; and a pair of immovable, horn-like antennules. However, many details relating to limb morphology, potentially important for phylogeny, have not been studied previously. Based on size categories, five or six larval stages can be recognized. The larvae approximately double their length and width during development (length: 230-520 microm). Most morphological features stay largely unchanged during development, but the antennal coxal masticatory spines are significant exceptions: they become bifid after one of the first molts. In all larval stages only the antennae and the mandibles actively move. In late naupliar stages the trunk limbs become visible as rows of laterally placed, undeveloped, and still immovable lobes. Swimming is performed by the antennae, whereas the mandibles appear to be involved mainly in feeding, as in other branchiopod larvae. The last naupliar stage undergoes a small metamorphosis to the first juvenile stage, the details of which in part were studied by following the premolt juvenile condition through the cuticle of the last stage nauplius. Among other changes there is a characteristic change in the shape and morphology of the univalved dorsal naupliar shield to a bivalved juvenile carapace. The general morphologies of the antennae and the mandibles are very similar to those of other branchiopod larvae and fall well within the "branchiopod naupliar feeding apparatus" recognized as a branchiopod synapomorphy by Olesen (2003), but some specific features shared with the larvae of other "conchostracans" are also identified. These special "conchostracan" features include: 1) a similar antennular setation; 2) a similar comb-like setulation of the bifid antennal coxal processes; and 3) mandibular palpsetae with setules condensed. In light of recent suggestions concerning branchiopod phylogeny (Cyclestheria as a sister group to the Cladocera), these similarities probably do not support a monophyletic "Conchostraca" but rather are symplesiomorphies of this taxon. A final decision must await a phylogenetic analysis of a more complete set of characters.

Animals↗

A 48-h larval development toxicity test using the marine polychaete Galeolaria caespitosa Lamarck (Fam. Serpulidae).

Assessing the risk of effluents and other anthropogenic inputs to the receiving environment is ultimately best done on a site-specific basis, which often requires toxicity tests using organisms relevant to that environment. Additionally, the test species or life stage needs to be available for a reasonable portion of the year to allow temporal fluctuations to be assessed. A 48-h larval development toxicity test using the marine polychaete Galeolaria caespitosa was developed. This test was developed as G. caespitosa releases viable gametes year-round, and the test species is environmentally relevant to the marine system receiving the liquid effluent being evaluated. Toxicity tests were conducted using G. caespitosa from different locations and evaluating the gamete response to copper. All population responses were comparable, with EC50 values ranging from 16 to 40 microg/L copper (as CuCl2, x 2H2O). Toxicity tests were also conducted using G. caespitosa gametes with an effluent produced by a lead smelting operation. The response of the G. caespitosa test with this effluent was compared with three other test methods using two microalgal species, Isochrysis sp. and Nitzschia closterium, and gametes from the mussel Mytilus edulis. The G. caespitosa larval development toxicity test was the most sensitive test to the effluent, with EC50 values ranging from 1-23% effluent, while it ranked second in sensitivity to copper. This test could be applied to other common serpulids worldwide.

Animals↗

Analysis of forkhead and snail expression reveals epithelial-mesenchymal transitions during embryonic and larval development of Nematostella vectensis.

The winged helix transcription factor Forkhead and the zinc finger transcription factor Snail are crucially involved in germ layer formation in Bilateria. Here, we isolated and characterized a homolog of forkhead/HNF3 (FoxA/group 1) and of snail from a diploblast, the sea anemone Nematostella vectensis. We show that Nematostella forkhead expression starts during late Blastula stage in a ring of cells that demarcate the blastopore margin during early gastrulation, thereby marking the boundary between ectodermal and endodermal tissue. snail, by contrast, is expressed in a complementary pattern in the center of forkhead-expressing cells marking the presumptive endodermal cells fated to ingress during gastrulation. In a significant portion of early gastrulating embryos, forkhead is expressed asymmetrically around the blastopore. While snail-expressing cells form the endodermal cell mass, forkhead marks the pharynx anlage throughout embryonic and larval development. In the primary polyp, forkhead remains expressed in the pharynx. The detailed analysis of forkhead and snail expression during Nematostella embryonic and larval development further suggests that endoderm formation results from epithelial invagination, mesenchymal immigration, and reorganization of the endodermal epithelial layer, that is, by epithelial-mesenchymal transitions (EMT) in combination with extensive morphogenetic movements. snail also governs EMT at different processes during embryonic development in Bilateria. Our data indicate that the function of snail in Diploblasts is to regulate motility and cell adhesion, supporting that the triggering of changes in cell behavior is the ancestral role of snail in Metazoa.

Amino Acid Sequence↗