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Roles of Hroth, the ascidian otx gene, in the differentiation of the brain (sensory vesicle) and anterior trunk epidermis in the larval development of Halocynthia roretzi.

Otx genes are expressed in the anterior neural tube and endoderm in all of the chordates so far examined. In mouse embryos, important roles of otx genes in the brain development have been well documented. However, roles of otx genes in other chordate species have been less characterized. To advance our understanding about roles of otx genes in chordates, we have studied Hroth, otx of the ascidian, Halocynthia roretzi. Hroth is expressed in the anterior part of the neural tube (the sensory vesicle), the endoderm and anterior epidermis in the development. In this study, we investigated roles of Hroth in the larval development through an antisense morpholino oligonucleotides (MOs) approach. Embryos injected with Hroth-targeting MO (Hroth knockdown embryos) developed into larvae without the adhesive organ, sensory pigment cells and cavity of the sensory vesicle. The tissues, in which defects were observed, are derived from anterior-animal cells of the embryo in early cleavage stages. During cleavage stages, Hroth is also expressed in the endoderm precursors of the vegetal hemisphere. However, Hroth expression in the anterior endoderm precursors do not seem to be essential for the above defects, since MO injection into the anterior-animal but not anterior-vegetal pair cells at the 8-cell stage gave the defects. Analysis of marker gene expression demonstrated that the fate choice of the sensory vesicle precursors and the specification of the sensory vesicle territory occurred normally, but the subsequent differentiation of the sensory vesicle was severely affected in Hroth knockdown embryos. The anterior trunk epidermis including the adhesive organ-forming region was also affected, indicating that anterior epidermal patterning requires Hroth function. Based on these findings, similarities and differences in the roles of otx genes between ascidians and mice are discussed.

Animals↗

[The larval development of Dipetalonema rugosicauda (syn. Wehrdikmansia rugosicauda) in the tick Ixodes ricinus. II. The development of Dipetalonema rugosicauda in Ixodes ricinus and investigations about the occurrence of the microfilariae in the roe deer (C. capreolus) (author's transl)].

Microfilariae of D. rugosicauda, which were taken up by nymphs of I. ricinus during their attachment on roe deers, developed in those ticks to infective larvae after two moults. The larval development lasted 56-67 days at 20-22 degrees C and occurred similar to other species of Dipetalonema inclusive a sausage stage. During this time the nymphs moulted to adult ticks. Adult females and males of I. ricinus collected in the area of Biberach/Riss, Southern-Germany, contained up to 12 infective larvae of D. rugosicauda per tick. The occurrence of those infective larvae and of microfilariae in roe deers depended on the season. The highest rate of infected ticks was 4.3% in spring, whereas the maximum rate of infected roe deers was 46.8% in autumn. Based on investigations of fawns of roe deers the prepatent period of D. rugosicauda can be estimated as approximately 6 months. Compared with microfilariae of Onchocerca tarsicola from red deers the microfilariae of D. rugosicauda prefered deeper layers of the ear skin of their hosts. Besides D. rugosicauda microfilariae of a further hitherto unknown species were detected in the ear skin of roe deers additionally. They resembled microfilariae of O. tarsicola.

Animals↗

Effects of sublethal exposure to Bacillus thuringiensis var. israelensis on larval development and adult size in Aedes aegypti.

The effects of exposure to sublethal concentrations of Bacillus thuringiensis var. israelensis (Serotype H-14) on second instar Aedes aegypti larvae were investigated. A test system was developed in which adverse effects would be detected as increased duration of larval development and decreased adult body size. No evidence of negative effects on survivors could be detected when sufficient B.t.i. dosages were applied to kill approximately half of the larvae in the treatment groups. However, when larval density was not controlled, and competition for food decreased as a result of larval mortality in the B.t.i.-treated groups, adult wing length was greater in the B.t.i. survivors than in the untreated controls. In addition, a residual mortality was noted in larvae that had been exposed to B.t.i. for 24 hr and then removed to habitats without B.t.i.

Aedes↗

Larval development and metamorphosis of the American lobster Homarus americanus (Crustacea, Decapoda): effect of eyestalk ablation and juvenile hormone injection.

Removal of eyestalks of Homarus americanus on different days and molting stages during larval development revealed that eyestalk tissue is involved in the larval and postlarval molting rhythm and in preparation for metamorphosis as early as the end of Stage II. Eyestalk removal in stages II and III reduced the duration of larval and postlarval stages. Eyestalk removal up to the end of Stage II delayed the completion of metamorphosis by one or two molts and caused additional development stages (designated IVa, IV', and V'). In this study, the critical stage for eyestalk ablation to delay metamorphosis occurred at the end of molt stage D1 of larval Stage II (the seventh day of development at 20 degrees). Injection of juvenile hormone before the critical stage resulted in a few intermediate stage IV' animals. This study demonstrates the involvement of eyestalk neuroendocrine tissue in the control of metamorphosis and investigates a possible involvement of juvenile hormone.

Animals↗

Effects of temperature on the larval development of Angiostrongylus cantonensis in the intermediate host, Biomphalaria glabrata.

The effects of temperature on the larval development of Angiostrongylus cantonensis in Biomphalaria glabrata were studied under controlled conditions. When B. glabrata were maintained at different, constant temperatures, the first-stage larvae developed to third-stage larvae between 20 degrees and 31 degrees C. The velocity of development to the third-stage in the snail depended on the relationship, Y = -0.1281 + 0.0081 X (Y velocity of development; X rearing temperature). The "threshold of development" was 15.8 degrees C and the thermal constant 123 degree-days. It is concluded that there might be a possibility of A. cantonensis spreading to temperate regions in the world, if, in addition to the temperature, other factors allowed the completion of the life cycle of this worm.

Angiostrongylus↗

The influence of an insect growth regulator on the larval development of the mud crab Rhithropanopeus harrisii.

Effects of the juvenile hormone analogue (S)-methoprene on the larval development and survival of the estuarine mud crab Rhitrhopanopeus harrisii were examined in the laboratory. Crab leave continuously exposed to 1000 microg (S)-methoprene litre(-1) did not survive beyond zoeal stage I. With continuous exposure to 100 microg (S)-methoprene litre(-1), mortality significantly increased through all larval stages except for zoeal stage II, and developmental duration significantly increased through all zoeal stages but not the premetamorphic megalopa. No supernumerary larval stages occurred with exposure (S)-methoprene. Rhithropanopeus harrisii larvae appear to be more sensitive to the single isomer formulation, (S)-methophrene, than to the double isomer formulation, (S)-methoprene, used in previously documented studies.

Journal Article↗

Experimental evidence of food-independent larval development in endemic Jamaican freshwater-breeding crabs.

In an experimental study, we compared reproductive and developmental traits of endemic sesarmid crabs from Jamaica living in landlocked limnic or terrestrial habitats. Laboratory rearing and behavioral observations showed that the larval development of Sesarma windsor, Sesarma dolphinum (both from freshwater brooks), and Metopaulias depressus (the bromeliad crab) invariably consists of two nonfeeding zoeal stages and a facultatively lecithotrophic megalopa. In a quantitative study of life-history processes characterizing this developmental mode, we provide for S. windsor first data of biomass and elemental composition (dry mass, W; carbon, hydrogen, nitrogen, collectively, CHN) during development from the egg through successive larval stages. These data show that larval independence of food is based on an enhanced female energy allocation in reproduction, reflected in unusually large egg size (1.45-1.70 mm), as well as high contents of C and H (about 60% and 9%, respectively) and high C : N ratios (7.6-8.4) in eggs and early larvae. During zoeal development, about 6% of initial W and 9% of N but 13% each of C and H were lost; similar losses occurred during megalopal development in continued absence of food. These patterns reflect the metabolic utilization of stored organic matter, with preferential degradation of lipid reserves. Fed megalopae gained greater amounts of W and N as compared with C and H (increments of 37% and 38% vs. 25% and 19%, respectively), indicating preferential investment of nutritional energy in proteins required for the formation of new tissues and organs, while generally decreasing proportions of CHN within total W suggested an increasing mineralization of the exoskeleton. Although survival and molt cycle duration of the megalopa stage were not affected by absence of food, significant effects were found in the size of first-stage juvenile crabs, indicating a trade-off between nutritional flexibility in the last larval stage and postmetamorphic fitness. Similar patterns of development and biomass in M. depressus as well as preliminary data obtained for S. dolphinum and Sesarma fossarum suggest that reproductive and developmental traits may be similar in all endemic Jamaican sesarmids. These traits are interpreted as life-history adaptations to development in landlocked habitats, probably playing a key role during adaptive radiation.

Adaptation, Physiological↗

The effect of repeated blood meals on the larval development of Dirofilaria corynodes in the fat body of Aedes aegypti.

The objective of the study was to determine whether blood meals taken by a vector subsequent to the infective one have any influence on the course of development of a filarial parasite in its tissue. The filaria Dirofilaria corynodes (von Linstow, 1899) was selected for study and the black-eye strain of Aedes aegypti served as the experimental vector. The microfilaria of D. corynodes develops to the infective stage in the fat bodies of the vector. Following an infective blood meal, mosquitoes were maintained on 10% sugar water alone (group A) or received supplementary blood meals i.e., an uninfected blood meal prior to the first larval molt followed by another after it (group B), an early blood meal (group C) or a late blood meal (group D). It was observed that additional blood meals after the infective one enhance the rate of larval development regardless of the timing of the feedings. An early blood meal, even if it is the only one, has a long-term significant effect on development. Repeated blood meals do not have an additive effect but tend to result in greater uniformity in larval size. A late blood meal also has a positive effect on development although the larval filariae do not grow or mature as rapidly as in the groups already mentioned. Larvae from mosquitoes fed on sugar water alone tended to develop at a slower rate and with less uniformity. It is concluded that infected mosquitoes taking one or more additional blood meals will yield a larger number of infective larvae in a shorter period of time.

Aedes↗

Cadmium-induced metallothionein expression during embryonic and early larval development of the mollusc Crassostrea virginica.

Newly fertilized eggs of the oyster Crassostrea virginica were exposed to 0.2 microM Cd and sampled during the first 24 hr of embryonic and larval development for determination of the temporal patterns of total bioaccumulated and metallothionein (MT)-bound Cd concentrations and the concentrations of MT mRNA. In comparison with controls, exposure to this concentration of Cd resulted in delayed development to the D-stage veliger larval stage. Maternal MT mRNA, which was carried over into the egg, declined immediately after fertilization. In controls, levels of MT mRNA were stable after this initial decline, and little or no detectable Cd was bound to MTs. Accumulation of new MT mRNA was observed in the exposed group after 12 hr of exposure when increases due to induction by Cd had occurred. Total accumulated Cd concentrations, which ranged from 0.04 to 0.1 pmol Cd/microg DNA in unexposed individuals, increased to 13.5 pmol Cd/microg DNA after 24 hr in Cd-exposed individuals. There was an increase in MT-bound Cd in exposed individuals prior to MT induction, which may be explained in part as Cd binding to maternal MT. This was detected as binding to CvNAcMT, the expected N-acetylated form of the oyster MT. Amounts of Cd bound to this form of MT increased, thereafter, and was followed by increases in binding to CvMT, the unexpected, nonacetylated form of the oyster MT, which has been associated with Cd toxicity, possibly in relation to the effects of Cd on normal cotranslational processing.

Animals↗

Expression of anterior Hox genes during larval development of the gastropod Haliotis asinina.

We report the spatial expression patterns of five anterior Hox genes during larval development of the gastropod mollusc Haliotis asinina, an unsegmented spiralian lophotrochozoan. Molecular alignments and phylogenetic analysis indicate that these genes are homologues of Drosophila HOM-C genes labial, proboscipedia, zen, Deformed, and Sex combs reduced; the abalone genes are named Has-Hox1, -Hox2, -Hox3, -Hox4, and -Hox5. Has-Hox transcripts are first detected in the free-swimming trochophore larval stage and restricted to the posttrochal ectoderm. Has-Hox2, -Hox3, and -Hox4 are expressed in bilaterally symmetrical and overlapping patterns in presumptive neuroectodermal cells on the ventral side of the trochophore. Has-Hox1 expression is restricted to a ring of cells on the dorsoposterior surface, corresponding to the outer mantle edge where new larval shell is being synthesized. There appears to be little change in the expression domains of these Has-Hox genes in pre- and posttorsional veliger larvae, with expression maintained in ectodermal and neuroectodermal tissues. Has-Hox2, -Hox3, -Hox4, and-Hox5 appear to be expressed in a colinear manner in the ganglia and connectives in the twisted nervous system. This pattern is not evident in older larvae. Has-Hox1 and-Hox4 are expressed in the margin of the mantle in the posttorsional veliger, suggesting that Hox genes play a role in gastropod shell formation.

Amino Acid Sequence↗

Evaluation of a larval development assay for the detection of anthelmintic resistance in Ostertagia circumcincta.

Two experiments were carried out to evaluate a larval development assay for the detection of anthelmintic resistance in O. circumcincta. In Experiment I, the dose responses to levamisole (LEV), thiabendazole (TBZ) and ivermectin (IVM) of 8 isolates of O. circumcincta were measured 34 days after infection (DAI). Four of these isolates were shown to be resistant to 1 or more anthelmintics. With 2 exceptions, all isolates considered to be resistant had higher LD50 values than the susceptible isolates for that anthelmintic. One exception was isolate RM8, which was considered to be resistant to all 3 anthelmintics based on faecal egg count reduction tests in goats, but the LD50 value for LEV did not differ from that for the susceptible isolates. The other exception was an isolate considered to be susceptible to TBZ which had a relatively high LD50 value. In an unrelated trial that was prompted by this finding, this isolate was confirmed to be benzimidazole-resistant. Isolate RM8 and an isolate susceptible to all 3 anthelmintics (SK2) were used in the second experiment, which was conducted to monitor changes in the LD50 values of LEV, TBZ and IVM over time following a single infection of 35,000 infective larvae in young sheep. Faecal samples were collected weekly from 24 to 115 DAI. With all 3 anthelmintics, the LD50 values increased with time to a peak around 50-60 DAI, and then declined to levels similar to those observed soon after patency. This trend was consistent for both isolates. The highest mean LD50 values for isolates SK2 for IVM and TBZ and RM8 for IVM and RM8, respectively, were 1.7 and 1.8 times, and 2.2 and 2.9 times higher than the initial mean LD50 values. There was a clear distinction in LD50 values between isolates at each sampling day for both IVM and TBZ. However, as a consequence of the changes in LD50 values with time, the peak LD50 values of IVM for isolate SK2 were higher than the minimum LD50 values of isolate RM8. As there was no apparent difference in LEV efficacy between these 2 isolates, the data were pooled. The highest mean LD50 value was 2.3 times higher than the initial LD50 value.

Animals↗

[The inotropic action of acetylcholine on the heart ventricles during larval development in the frog Rana temporaria].

The effects of acetylcholine and other cholinergic drugs on the isolated electrically driven larval frog ventricles have been studied. The negative inotropic response to acetylcholine appeared as early as stage 33 of the larval development (the stages were determined according to Dabagian and Sleptsova, 1975) and persisted through all the developmental stages including metamorphosis. The response is muscarinic in origin since it was reproduced with a muscarinic agonist methylfurmetide, blocked with atropine but was not modified with tubocurarine. At the stage 41 and following stages, the sensitivity to acetylcholine was decreased while to methylfurmetide was not. The decreased sensitivity to acetylcholine is most likely due to increase of activity of cholinesterases in the myocardial tissues.

Acetylcholine↗

[Effects of nutrient and salinity in egg and larval development of Aedes togoi].

The study made an observation on periodicity of oviposition, and the effects of nutrient and salinity in egg and larval development of Aedes togoi, and the results are summarized as follows: The 53.9% mosquitoes of one feeding laid eggs once, 26.9% laid twice and 19.2% laid three times. Autogenous rate of Aedes togoi reared in three different nutrient groups in larval sage was 6.9% in 0.8 mg/larva, 22.5% in 1.6 mg/larva and 44.4% in 2.4 mg/larva. The oviposition rate according to different salinity of the oviposition sites (0%, 0.5%, 1.0%, 2.0% and 4.0%) was 25.2% in distilled water, 36.2% in 0.5% salinity, 23.5% in 1.0% salinity and 14.6% in 2.0% salinity. Developing period of the aquatic stage of male Aedes togoi in 25 degrees C were shorter (10.73 days) than females (11.85 days). The most effective concentration of salinity for the developing period was 1.0% which took 9.25 days in males and 10.44 days in females. In the developmental status of the follicles according to nutrition in the larval stage, the numbers of follicles of groups fed 0.8 mg, 1.6 mg and 2.4 mg per larva were 180.7, 197.5 and 202 respectively. The result of ovary dissection on the 10th day after emergence, three different nutrition groups were in Christopher's stage IIa mostly; each 71.0%, 61.1% and 39.9% of the total follicles and autogenous females observed.

Animal Nutritional Physiological Phenomena↗

[The larval development of the warble-fly Oestromyia leporina Pall. (Deptera, Hyodermah'dae) (author's transl)].

The present paper deals with the larval development of the warble-fly Oestromyia leporina and especially with the development of the first instar larvae (L 1). The hatching from the egg and the modus of infection is described. The migration of the L 1 within the host is reported for the first time as well as the perforation of the skin after the migration period. Exact dates on the duration of the different larval stages are given. Experimental infection reveals a partial immunization of the host against new infections. Some remaining problems of larval biology are discussed. By comparison several facts suggest our present knowledge of the perforation of larval (L 1) Hypoderma bovis probably to be wrong.

Animals↗

The Drosophila melanogaster stranded at second (sas) gene encodes a putative epidermal cell surface receptor required for larval development.

Several lethal mutations were identified previously in the 84BD interval of the Drosophila melanogaster third chromosome (Lewis et al., 1980; Cavener et al., 1986b). We have examined the l(3)84Cd complementation group and found that mutants exhibit novel cuticular defects and die during larval development. The lethal phase occurs during the first larval molt or subsequently during the second instar larval stage; hence, we have named the gene stranded at second (sas). There are no apparent effects on the rate of development of embryos or first instar larvae. Second instar larvae which survive the molt exhibit a marked reduction in growth and eventually die as small second instar larvae. Incomplete penetrance in some weak sas alleles can yield fertile adults. In addition to the lethal phenotype, a segmentally repeated pattern of tanned spots is found within the ventral setal belts of mutant larvae. The position of the spots is always either between the fourth and fifth row of setae (cuticular projections) or between the first and second row of setae. The spots are adjacent to the muscle attachment sites in the setal belt region. Another common larval phenotype is the abnormal tanning of the ventral surface of the pharynx. The sas gene was cloned, and both the cuticular tanning and the larval lethal phenotypes were complemented by P-element-mediated transformation with a genomic DNA-cDNA construct. Three major sas transcripts are expressed throughout development in cuticle secreting epidermal tissues. The sas transcripts show stage- and tissue-specific patterns of expression with switches in transcript patterns occurring at the molts. The inferred 1348-amino-acid sequence suggests that sas encodes a cell surface protein which functions as a receptor. The putative extracellular region contains four tandem repeats of a cysteine-rich motif which is similar to a cysteine pattern present in procollagen and in thrombospondin. Following this region are at least three copies of a fibronectin type III class repeat. The short (35 amino acids) intracellular domain contains a sequence (NPXY) that has been implicated in endocytosis via coated pits.

Amino Acid Sequence↗

Differentiation of two locally sympatric Protopolystoma (Monogenea: Polystomatidae) species by temperature-dependent larval development and survival.

The developmental response of egg stages to different environmental temperature regimes was studied in Protopolystoma xenopodis and Protopolystoma orientalis (Monogenea: Polystomatidae) isolates from southern Africa. Eggs failed to develop at 10 degrees C, whilst at 15 degrees C only P. xenopodis completed larval development, hatching 49--88 days post-collection. Respective hatching windows were 26--34 (P. xenopodis) and 37--49 (P. orientalis) days at 20 degrees C, and 18--26 and 27--37 days at 25 degrees C. Continuous maintenance at 30 degrees C was lethal for eggs of both species. There were no consistent interspecific differences in the response of egg stages to low and high temperature shocks during early embryonic development.

Africa, Southern↗

Seawater type shapes larval development, intestinal microbiota, and water quality in Macrobrachium rosenbergii.

Giant freshwater prawn (Macrobrachium rosenbergii) has high economic value and extensive aquaculture prospects, and larval quality critically restricts the sustainable development of the industry. To investigate the effects of different seawater sources on larval survival and growth, artificial seawater, natural seawater from Beibu Gulf (China) and East China Sea (China) were used as culture media to compare their influences on larval development and intestinal microbiota of M. rosenbergii. Significant intergroup differences were observed in cultivation outcomes. The larval emergence rate reached the maximum value of 52.1% in Group G (Beibu Gulf natural seawater), followed by 43.57% in Group R (artificial seawater), and only 36.4% in Group Z (East China Sea natural seawater), indicating distinct larval emergence gaps and a gradual decline in larval development efficiency. Intestinal microbiota analysis indicated that Pseudomonadota, Actinomycetota and Bacillota were the dominant phyla in all three groups. The relative abundance of Bacillota increased significantly in Group G on day 10. LEfSe analysis proved that this group owned the most stable microbial structure and highest community diversity, and pronounced structural fluctuations occurred in Group R, suggesting that intestinal microbial profiles can reflect larval developmental conditions. Correlation analysis showed that gut microbial composition was closely correlated with water environmental parameters, and microbial metabolic pathways were strongly associated with larval growth and development. This study indicates that natural seawater from the Beibu Gulf of China is more suitable for healthy larval cultivation of M. rosenbergii. Structural stability and diversity of intestinal microbiota are key factors affecting larval growth. The results offer theoretical references for optimizing artificial seawater formula and improving larval rearing efficiency.

16S rRNA↗

Characterization of a partial alpha-amylase clone from red porgy (Pagrus pagrus): expression during larval development.

A partial alpha-amylase cDNA was isolated from red porgy (Pagrus pagrus, Teleostei: Sparidae) and its tissue specific expression during larval development was examined. The cDNA was 949 bp long and showed 90% identity with other fish amylases. A 545 bp fragment was used to study amylase expression using in situ hybridization and RT-PCR techniques. Both methods showed a similar pattern: high and relatively constant expression for the first 30 days after hatching (dah), subsequently decreasing until the end of the experiment at 60 dah. The goal of this work was to extend the existing knowledge of the functionality of larval fish digestive systems and to provide new information about alpha-amylase gene expression.

Age Factors↗