The megalopa and juvenile development of Pachygrapsus transversus (Gibbes, 1850) (Decapoda, Brahyura) compared with other grapsid crabs

Megalopae were reared in the laboratory to the 7(th) crab stage. The megalopa and 1(st) crab stage are described and juvenile development was studied with emphasis on pleopodal differentiation. The megalopal phase, is easily identified, and shares with those of other Grapsinae and Plagusiinae big size, the presence of many natatory setae, and a series of conspicuous teeth on the inner margin of the dactyli from the 2(nd) to 4(th) walking leg. These features are regarded as adaptive for settlement in a wave-swept environment, such as the rocky marine intertidal where most of those species live. Fast development of juvenile pleopods is another characteristic of these subfamilies. In Pachygrapsus transversus, the sexes can be distinguished from the 2(nd) crab stage. Gonopod differentiation in males and the basic segmentation of all four pleopod pairs in females are already concluded at the 5(th) instar. A review of the available information indicated that settlement of large megalopae and fast juvenile development, preceding a precocious sexual maturity, are trends in Grapsinae and Plagusiinae. on the other hand, the Sesarminae pass through a more extensive juvenile instar sequence and presumably a delayed maturity.

Effect of the juvenile hormone on the development of the mandibular gland in workers' pupae of Apis mellifera L. (Hymenoptera, Apidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Audouinella macrospora (Acrochaetiaceae, Rhodophyta) is the Chantransia stage of Batrachospermum (Batrachospermaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Growth and juvenile development of Uca maracoani Latreille 1802-1803 in laboratory conditions (Crustacea, Decapoda, Brachyura, Ocypodidae).

The life cycle of decapod crustaceans can be classified into three distinct morphological phases: larval, juvenile and adult. Despite its recognized importance, studies of the juvenile phase have been neglected. The present Study aimed to analyze the growth of juveniles from a single population of Uca maracoani under laboratory conditions, and also to describe the morphological differentiation of pleopods in each sex. Megalopae and juvenile crabs or U. maracoani obtained on a Mud beach at Jabaquara, Paraty, on the southern coast of the state of Rio de Janeiro (Brazil), were reared in the laboratory. The specimens were checked daily for molts and deaths. The carapace widths (CW) of intact exuviae and dead individuals were measured under a stereoscopic microscope provided with a micrometer rule. These data allowed the definition of a growth equation as well as the stages related to the beginning of pleopod development, which begins when females reach 3.0 mill CW (6th juvenile developmental stage), similar to the sizes reported for other species of the genus. In males, however, pleopods appear when the crabs reach 3.5 mm CW, equivalent to the 7th developmental stage. This difference may be related to differential growth between sexes. It also may be a consequence of laboratory rearing, or may represent an actual feature of the species.

Somatic meiosis and development of the juvenile gametophyte in members of the Batrachospermales sensu lato (Rhodophyta)

Seven populations (six in culture and one sampled directly from nature) of the freshwater red algal families Batrachospermaceae, Lemaneaceae and Thoreaceae were examined, involving three species of Batrachospermum, two of Paralemanea and one of Thorea. All 'Chantransia' stages ultimately produced juvenile gametophytes. The production of juvenile gametophytes in the three populations of Batrachospermum was generally most abundant at 15°C and low irradiances (47-68 μmol photons m-2 s-1). The most abundant gametophyte development in the Paralemanea species was observed at 10°C and low or high irradiances (47-142 μmol photons m-2 s-1). Gametophyte production in Thoreaceae occurred at higher temperatures (20°C) and also at low irradiances. In species of the Batrachospermaceae and Lemaneaceae, the 'elimination cells' can be situated on the basal or suprabasal cell of the juvenile gametophyte, but the position is usually fixed in individual species. The presence and position of the elimination cells remain to be established in Thoreaceae. Our results corroborate a previous study suggesting that the position of elimination cells is such a constant feature that it is of potential diagnostic value at the generic or infrageneric (sectional or specific) level. The characteristics observed in the development of the juvenile gametophytes in species of Batrachospermaceae and Lemaneaceae essentially agreed with general descriptions in the previous studies. The characteristics of the Thoreaceae, with a distinctive developmental pattern of the juvenile gametophyte and the occurrence of two morphological types in the 'Chantransia' stage, support the proposal to elevate it to the ordinal level. Two remarkable observations in Batrachospermum species were the production of numerous juvenile gametophytes from filaments of the same plant of the 'Chantransia' stage and the formation of a system of rhizoidal filaments or cell agglomeration of the juvenile gametophytes, which produced new gametophytes. These two characteristics potentially increase the formation of additional gametophytes under favourable conditions.

The megalopa stage of Portunus spinimanus Latreille, 1819 and Portunus gibbesii (Stimpson, 1859) (Decapoda, Brachyura, Portunidae) from the southeastern Atlantic coast of the United States

The identification of megalopae from plankton samples is difficult, because this larval stage is the least well known among crab larvae, unknown in some species and poorly described in others. Wild megalopa specimens of some swimming crabs (family Portunidae Rafinesque, 1815) were captured alive from neuston samples obtained during summer surveys near the coast of Charleston, South Carolina (U.S.A). For identification purposes, larvae were reared to the 8th juvenile instar. After reaching the 5th juvenile instar, the juvenile crabs exhibited morphological features suitable for identification to the species level. The specimens belonged to two species of Portunidae, Portunus spinimanus Latreille, 1819 and P. gibbesii (Stimpson, 1859). Their megalopae were described in detail and compared to other portunid megalopae known from the southeastern Atlantic coast of the U.S.A. Species-specific characters of portunid megalopae are the number of carpal spines on the chelipeds, the relative size of the sternal spines (7th sternite), the number of antennal flagellum segments, and the setation of mouthparts. Copyright © 2007 Magnolia Press.

Effects of treatment of the fat body trophocytes of Melipona quadrifasciata anthidioides nurse workers and virgin queens in culture by juvenile hormone III and ecdysterone (20-HE)

The fat body (FB) consists of two types of cells: throphocytes and oenocytes. Throphocytes are related to intermediary metabolism storing lipids, carbohydrates, and proteins while oenocytes play role in the lipids and lipoproteins production. The vitellogenin is the precursor of egg yolk (vitelline) and is synthesized on FB. The aim of this work was to analyze the effects of hormones acting in bee reproduction, as juvenile hormone (JH) and ecdisteroids (20 HE) on FB cells, where vitellogenin is synthesized. For the study were chose nurse workers that in Melipona quadrifasciata anthidioides present activated ovaries and produce eggs, and virgin queens whose ovaries are not yet activated, presenting only previtellogenic follicles. FB trophocytes from these classes of bees were cultivated in media containing different amounts of JH and 20-HE. The effects on trophocytes cytoplasm reserves of lipids, proteins, and activity of acid phosphatase were compared by observing preparations from cultured FB, treated and control, by transmission electron microscopy (TEM). The results showed that the hormones effects are related to the bee's caste and functional ovary stage. The role of acid phosphatase on mobilization of the trophocyte reserves was also determined. © 2012 Wiley Periodicals, Inc.

Redescription of megalopa and juvenile development of Pachygrapsus gracilis (Decapoda: Grapsidae) from the Amazon region, reared in the laboratory

ABSTRACT: This present study re-describes the megalopa stage and provides detailed morphological descriptions and main growth changes observed in stages I through VII of the juvenile instars of the dark shore crab Pachygrapsus gracilis (Saussure, 1858), from the Amazon region. The specimens in this study were reared in the laboratory and the megalopae were collected at Ajuruteua beach in northeastern Pará, Brazil. Previous studies had described the megalopa of P. gracilis from Mexican waters, as well as those of Pachygrapsus transversus (Gibbes, 1850) and Pachygrapsus marmoratus (Fabricius, 1787). A comparison between the Mexican and Amazonian populations of P. gracilis revealed significant morphological differences. The main difference is the presence of 3 elongated setae on the 5th pereiopods of individuals of the Amazonian population. The setal number and their arrangement in the appendages also differed. In P. gracilis, the male and female genital openings are observed from the juvenile instar III, whereas differentiation in male pleopods is observed only in juvenile instar V. In females, the pleopods undergo rapid differentiation during juvenile instar VI. These morphological comparisons and other observations on development are briefly compared and discussed with reports for other species.

Molting dynamics and juvenile hormone titer profiles in the nymphal stages of a lower termite, Cryptotermes secundus (Kalotermitidae) - signatures of developmental plasticity

Termites are social cockroaches and this sociality is founded on a high plasticity during development. Three molting types (progressive, stationary and regressive molts) are fundamental to achieve plasticity during alate/sexual development, and they make termites a major challenge to any model on endocrine regulation in insect development. As the endocrine signatures underpinning this plasticity are barely understood, we studied the developmental dynamics and their underlying juvenile hormone OH) titers in a wood-dwelling termite. Cryptotermes secundus, which is characterized by an ancestral life style of living in dead wood and individuals being totipotent in development. The following general pattern elements could be identified during winged sexual development (i) regressive molts were accompanied by longer intermolt periods than other molting types, (ii) JH titers decreased gradually during the developmental transition from larva (immatures without wing buds), to nymph (immatures with wing buds), to winged adult, (iii) in all nymphal stages, the JH titer rose before the next molt and dropped thereafter within the first week, (iv) considerable variation in JH titers occurred in the midphase of the molting cycle of the 2nd and 3rd nymphal instar, inferring that this variation may reflect the underlying endocrine signature of each of the three molting types, (v) the 4th nymphal instar, the shortest of all, seems to be a switch point in development, as nymphs in this stage mainly developed progressively. When comparing these patterns with endocrine signatures seen in cockroaches, the developmental program of Cryprotermes can be interpreted as a co-option and repetitive use of hormonal dynamics of the post dorsal-closure phase of cockroach embryonic development. (C) 2012 Elsevier Ltd. All tights reserved.

The endocrine basis for reproductive life-history trade-offs during the previtellogenic resting stage in the yellow fever mosquito, Aedes Aegypti

Juvenile hormone (JH) is the central hormonal regulator of life-history trade-offs in many insects. In Aedes aegypti, JH regulates reproductive development after emergence. Little is known about JH's physiological functions after reproductive development is complete or JH's role in mediating life-history trade-offs. By examining the effect of hormones, nutrition, and mating on ovarian physiology during the previtellogenic resting stage, critical roles were determined for these factors in mediating life-history trade-offs and reproductive output. The extent of follicular resorption during the previtellogenic resting stage is dependent on nutritional quality. Feeding females a low quality diet during the resting stage causes the rate of follicular resorption to increase and reproductive output to decrease. Conversely, feeding females a high quality diet causes resorption to remain low. The extent of resorption can be increased by separating the ovaries from a source of JH or decreased by exogenous application of methoprene. Active caspases were localized to resorbing follicles indicating that an apoptosis-like mechanism participates in follicular resorption. Accumulations of neutral lipids and the accumulation of mRNA's integral to endocytosis and oocyte development such as the vitellogenin receptor (AaVgR), lipophorin receptor (AaLpRov), heavy-chain clathrin (AaCHC), and ribosomal protein L32 (rpL32) were also examined under various nutritional and hormonal conditions. The abundance of mRNA's and neutral lipid content increased within the previtellogenic ovary as mosquitoes were offered increasing sucrose concentrations or were treated with methoprene. These same nutritional and hormonal manipulations altered the extent of resorption after a blood meal indicating that the fate of follicles and overall fecundity depends, in part, on nutritional and hormonal status during the previtellogenic resting stage. Mating female mosquitoes also altered follicle quality and resorption similarly to nutrition or hormonal application and demonstrates that male accessory gland substances such as JH III passed to the female during copulation have a strong effect on ovarian physiology during the previtellogenic resting stage and can influence reproductive output. Taken together these results demonstrate that the previtellogenic resting stage is not an inactive period but is instead a period marked by extensive life-history and fitness trade-offs in response to nutrition, hormones and mating stimuli.

Hepatic Responses of Juvenile Fundulus heteroclitus from Pollution-adapted and Nonadapted Populations Exposed to Elizabeth River Sediment Extract.

Atlantic killifish (Fundulus heteroclitus) inhabiting the Atlantic Wood Industries region of the Elizabeth River, Virginia, have passed polycyclic aromatic hydrocarbon (PAH) resistance to their offspring as evidenced by early life stage testing of developmental toxicity after exposure to specific PAHs. Our study focused on environmentally relevant PAH mixtures in the form of Elizabeth River sediment extract (ERSE). Juvenile (5 month) F1 progeny of pollution-adapted Atlantic Wood (AW) parents and of reference site (King's Creek [KC]) parents were exposed as embryos to ERSE. Liver alterations, including nonneoplastic lesions and microvesicular vacuolation, were observed in both populations. ERSE-exposed KC fish developed significantly more alterations than unexposed KC fish. Interestingly, unexposed AW killifish developed significantly more alterations than unexposed KC individuals, suggesting that AW juveniles are not fully protected from liver disease; rapid growth of juvenile fish may also be an accelerating factor for tumorigenesis. Because recent reports show hepatic tumor formation in adult AW fish, the differing responses from the 2 populations provided a way to determine whether embryo toxicity protection extends to juveniles. Future investigations will analyze older life stages of killifish to determine differences in responses related to chronic disease.

Tegument Glycoproteins and Cathepsins of Newly Excysted Juvenile Fasciola hepatica Carry Mannosidic and Paucimannosidic N-glycans

Recently, the prevalence of Fasciola hepatica in some areas has increased considerably and the availability of a vaccine to protect livestock from infection would represent a major advance in tools available for controlling this disease. To date, most vaccine-target discovery research on this parasite has concentrated on proteomic and transcriptomic approaches whereas little work has been carried out on glycosylation. As the F. hepatica tegument (Teg) may contain glycans potentially relevant to vaccine development and the Newly Excysted Juvenile (NEJ) is the first lifecycle stage in contact with the definitive host, our work has focused on assessing the glycosylation of the NEJTeg and identifying the NEJTeg glycoprotein repertoire. After in vitro excystation, NEJ were fixed and NEJTeg was extracted. Matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) analysis of released N-glycans revealed that oligomannose and core-fucosylated truncated N-glycans were the most dominant glycan types. By lectin binding studies these glycans were identified mainly on the NEJ surface, together with the oral and ventral suckers. NEJTeg glycoproteins were affinity purified after targeted biotinylation of the glycans and identified using liquid chromatography and tandem mass spectrometry (LC-MS/MS). From the total set of proteins previously identified in NEJTeg, eighteen were also detected in the glycosylated fraction, including the F. hepatica Cathepsin B3 (FhCB3) and two of the Cathepsin L3 (FhCL3) proteins, among others. To confirm glycosylation of cathepsins, analysis at the glycopeptide level by LC-ESI-ion-trap-MS/MS with collision-induced dissociation (CID) and electron-transfer dissociation (ETD) was carried out. We established that cathepsin B1 (FhCB1) on position N80, and FhCL3 (BN1106_s10139B000014, scaffold10139) on position N153, carry unusual paucimannosidic Man2GlcNAc2 glycans. To our knowledge, this is the first description of F. hepatica NEJ glycosylation and the first report of N-glycosylation of F. hepatica cathepsins. The significance of these findings for immunological studies and vaccine development is discussed.

Stimulating Neoblast-Like Cell Proliferation in Juvenile Fasciola hepatica Supports Growth and Progression towards the Adult Phenotype In Vitro

Fascioliasis (or fasciolosis) is a socioeconomically important parasitic disease caused by liver flukes of the genus Fasciola. Flukicide resistance has exposed the need for new drugs and/or a vaccine for liver fluke control. A rapidly improving 'molecular toolbox' for liver fluke encompasses quality genomic/transcriptomic datasets and an RNA interference platform that facilitates functional genomics approaches to drug/vaccine target validation. The exploitation of these resources is undermined by the absence of effective culture/maintenance systems that would support in vitro studies on juvenile fluke development/biology. Here we report markedly improved in vitro maintenance methods for Fasciola hepatica that achieved 65% survival of juvenile fluke after 6 months in standard cell culture medium supplemented with 50% chicken serum. We discovered that this long-term maintenance was dependent upon fluke growth, which was supported by increased proliferation of cells resembling the "neoblast" stem cells described in other flatworms. Growth led to dramatic morphological changes in juveniles, including the development of the digestive tract, reproductive organs and the tegument, towards more adult-like forms. The inhibition of DNA synthesis prevented neoblast-like cell proliferation and inhibited growth/development. Supporting our assertion that we have triggered the development of juveniles towards adult-like fluke, mass spectrometric analyses showed that growing fluke have an excretory/secretory protein profile that is distinct from that of newly-excysted juveniles and more closely resembles that of ex vivo immature and adult fluke. Further, in vitro maintained fluke displayed a transition in their movement from the probing behaviour associated with migrating stage worms to a slower wave-like motility seen in adults. Our ability to stimulate neoblast-like cell proliferation and growth in F. hepatica underpins the first simple platform for their long-term in vitro study, complementing the recent expansion in liver fluke resources and facilitating in vitro target validation studies of the developmental biology of liver fluke.

Evidence from genetic and Lagrangian drifter data for transatlantic transport of small juvenile green turtles

[EN] Aim: A key life-history component for many animals is the need for movement between different geographical locations at particular times. Green turtle (Chelonia mydas) hatchlings disperse from their natal location to spend an early pelagic stage in the ocean, followed by a neritic stage where small juveniles settle in coastal areas. In this study, we combined genetic and Lagrangian drifter data to investigate the connectivity between natal and foraging locations. In particular, we focus on the evidence for transatlantic transport.

Evidence from genetic and Lagrangian drifter data for transatlantic transport of small juvenile green turtles

[EN] Green turtle hatchlings disperse away from their natal location to spend an early pelagic stage in the ocean, followed by a neritic stage where small juveniles settle in coastal areas. Here, we combined genetic and Lagrangian drifter data to investigate the connectivity between natal and foraging locations; particularly focussing on the evidence for transatlantic transport. Our results supported the general hypothesis that turtles tend to select foraging areas ‘closest-to-home’.