Larval Types Research Articles

Measuring rootworm refuge function: Diabrotica virgifera virgifera emergence and mating in seed blend and strip refuges for Bacillus thuringiensis (Bt) maize.

Current insect resistance management plans rely on refuges of plants without Bacillus thuringiensis (Bt) toxins to provide a gene pool of unexposed insects. Insects from refuges must mate with insects from Bt maize to slow resistance evolution. We used stable isotope labeling to observe Diabrotica virgifera virgifera emergence, dispersal, physical characteristics, and mating in Bt and refuge maize planted in different refuge configurations. Our objective was to assess how refuge type facilitates mating between insects from Bt and refuge plants. Mating between D. v. virgifera beetles from different plant types was more likely in seed blends compared with strip refuges. Adult D. v. virgifera from refuge plants emerged before those from Bt plants. In strip refuges, D. v. virgifera from refuge plants did not disperse far from refuge boundaries. Larval host plant type did not affect adult size. Larger males and females were more likely to mate. Low proportions of D. v. virgifera from refuge plants were found in 5% seed blend refuges. Seed blend refuges can help to facilitate gene flow between D. v. virgifera beetles from Bt and refuge maize, but current approaches do not meaningfully contribute to delaying resistance because numbers of refuge beetles produced are insufficient. © 2018 Society of Chemical Industry.

Cnidarian Cell Type Diversity and Regulation Revealed by Whole-Organism Single-Cell RNA-Seq

The emergence and diversification of cell types is a leading factor in animal evolution. So far, systematic characterization of the gene regulatory programs associated with cell type specificity was limited to few cell types and few species. Here, we perform whole-organism single-cell transcriptomics to map adult and larval cell types in the cnidarian Nematostella vectensis, a non-bilaterian animal with complex tissue-level body-plan organization. We uncover eight broad cell classes in Nematostella, including neurons, cnidocytes, and digestive cells. Each class comprises different subtypes defined by the expression of multiple specific markers. In particular, we characterize a surprisingly diverse repertoire of neurons, which comparative analysis suggests arethe result of lineage-specific diversification. By integrating transcription factor expression, chromatin profiling, and sequence motif analysis, we identify the regulatory codes that underlie Nematostella cell-specific expression. Our study reveals cnidarian cell type complexity and provides insights into the evolution of animal cell-specific genomic regulation.

New and known zoonotic nematode larvae within selected fish species from Queensland waters in Australia

Zoonotic fish-borne nematodes are a significant and growing global health concern. However, in Australia their occurrence and geographical distribution are not well known. Moreton Bay in Queensland, Australia, is a popular tourist destination for domestic and international visitors where seafood is a popular food item. The aim of this study was to determine the occurrence of zoonotic nematodes within four commercially significant teleost fish species from Moreton Bay; Surf Bream, Sea Mullet, Yellow-Fin Tripod fish and Gold-Lined Sea Bream. Overall, 34.03% of fish examined (n = 144) were found to be a host to at least one parasitic nematode. The seasonal dynamics and prevalence of nematode infection was tested by statistical means. Results indicated a significant difference in seasonal infection for Surf Bream (p < 0.005) only. Several potentially zoonotic larval morphotypes were found, including Contracaecum types I and II, and Terranova type II (Family Anisakidae), and Hysterothylacium types VI and XIV and a new Hysterothylacium larval type herein assigned as type XVII (Family Raphidascarididae). ITS sequence data showed that some Contracaecum type II and Hysterothylacium types VI found in the present study are genetically distinct from those previously reported. The new genotypes were assigned to Contracaecum type II Genotype B and Hysterothylacium type VI, Genotypes B and C. This study provides essential information for future research on specific identification and differentiation of infective stages of zoonotic nematoda in edible fish.

Molecular phylogenetic analysis of a taxonomically unstable ranid from Sumatra, Indonesia, reveals a new genus with gastromyzophorous tadpoles and two new species

The presence of an adhesive abdominal sucker (gastromyzophory) allows tadpoles of certain species of anurans to live in fast-flowing streams. Gastromyzophorous tadpoles are rare among anurans, known only in certain American bufonids and Asian ranids. To date,Huia sumatrana, which inhabits cascading streams, has been the only Sumatran ranid known to possess gastromyzophorous tadpoles. In the absence of thorough sampling and molecular barcoding of adults and larvae, it has remained to be confirmed whether other Sumatran ranid species living in similar habitats, i.e.,Chalcorana crassiovis, possesses this larval type. Moreover, the taxonomic status of this species has long been uncertain and its taxonomic position within the Ranidae, previously based exclusively on morphological characters, has remained unresolved. To study the diversity and relationships of these frogs and to establish the identity of newly collected gastromyzophorous tadpoles from Sumatra, we compared genetic sequences ofC. crassiovis-like taxa from a wide range of sites on Sumatra. We conducted bayesian and maximum likelihood phylogenetic analyses on a concatenated dataset of mitochondrial (12S rRNA, 16S rRNA, and tRNAval) and nuclear (RAG1 and TYR) gene fragments. Our analyses recoveredC. crassiovisto be related toClinotarsus,Huia, andMeristogenys. The DNA barcodes of the gastromyzophorous tadpoles matched adults from the same sites. Herein, we provide a re-description of adultC. crassiovisand propose “C. kampeni” as a synonym of this species. The molecular evidence, morphological features, and distribution suggest the presence of two related new species. The two new species andC. crassiovistogether represent a distinct phylogenetic clade possessing unique molecular and morphological synapomorphies, thus warranting a new genus.

From ground pools to treeholes: convergent evolution of habitat and phenotype in Aedes mosquitoes

BackgroundInvasive mosquito species are responsible for millions of vector-borne disease cases annually. The global invasive success of Aedes mosquitoes such as Aedes aegypti and Aedes albopictus has relied on the human transport of immature stages in container habitats. However, despite the importance of these mosquitoes and this ecological specialization to their widespread dispersal, evolution of habitat specialization in this group has remained largely unstudied. We use comparative methods to evaluate the evolution of habitat specialization and its potential influence on larval morphology, and evaluate whether container dwelling and invasiveness are monophyletic in Aedes.ResultsWe show that habitat specialization has evolved repeatedly from ancestral ground pool usage to specialization in container habitats. Furthermore, we find that larval morphological scores are significantly associated with larval habitat when accounting for evolutionary relationships. We find that Ornstein-Uhleinbeck models with unique optima for each larval habitat type are preferred over several other models based predominantly on neutral processes, and that OU models can reliably simulate real morphological data.ConclusionsOur results demonstrate that multiple lineages of Aedes have convergently evolved a key trait associated with invasive success: the use of container habitats for immature stages. Moreover, our results demonstrate convergence in morphological characteristics as well, and suggest a role of adaptation to habitat specialization in driving phenotypic diversity in this mosquito lineage. Finally, our results highlight that the genus Aedes is not monophyletic.

A multispecies approach reveals hot spots and cold spots of diversity and connectivity in invertebrate species with contrasting dispersal modes.

Genetic diversity is crucial for species' maintenance and persistence, yet is often overlooked in conservation studies. Species diversity is more often reported due to practical constraints, but it is unknown if these measures of diversity are correlated. In marine invertebrates, adults are often sessile or sedentary and populations exchange genes via dispersal of gametes and larvae. Species with a larval period are expected to have more connected populations than those without larval dispersal. We assessed the relationship between measures of species and genetic diversity, and between dispersal ability and connectivity. We compiled data on genetic patterns and life history traits in nine species across five phyla. Sampling sites spanned 600km in the northwest Mediterranean Sea and focused on a 50-km area near Marseilles, France. Comparative population genetic approaches yielded three main results. (i) Species without larvae showed higher levels of genetic structure than species with free-living larvae, but the role of larval type (lecithotrophic or planktotrophic) was negligible. (ii) A narrow area around Marseilles, subject to offshore advection, limited genetic connectivity in most species. (iii) We identified sites with significant positive contributions to overall genetic diversity across all species, corresponding with areas near low human population densities. In contrast, high levels of human activity corresponded with a negative contribution to overall genetic diversity. Genetic diversity within species was positively and significantly linearly related to local species diversity. Our study suggests that local contribution to overall genetic diversity should be taken into account for future conservation strategies.

Abundance and Dynamics of Anopheles (Diptera: Culicidae) Larvae in a Malaria Endemic Area of Bangladesh.

Malaria is a major health burden in the border-belt areas of Bangladesh. There are recent data from adult mosquito collections that a number of vectors are involved in the transmission cycle. However, little information regarding the larval habitats of Anopheles mosquitoes are available in Bangladesh. To start filling this gap, a cross-sectional larval survey was conducted in Bandarban, Bangladesh from October 2011 to September 2012. Descriptive analysis, Poisson regression, spearman correlations and zero-inflated Poisson regression were used to calculate the degree of association between the abundance of larval Anopheles species and environmental factors. From the 300 larval habitats sampled, 5,568 Anopheles larvae were collected and of these, 2,263 (40.6%) were identified to species. Collections represented 16 Anopheles species with Anopheles vagus (26.4%, n = 598) as the dominant species. A total of 16 Anopheles larval habitat types were identified. Larval abundance was significantly different (P < 0.05) among habitats with pond (40%, n = 914) and rice field (34%, n = 779) implicated to be the most utilized. Larval abundance varied significantly (P < 0.05) with habitat characteristics. Most of the larvae were collected from sites with a range of pH from 7.0 to 8.0. Data obtained from this study revealed both natural and human-created larval habitats were favorable for anopheline larval survival and development. Such information elucidates plausible drivers of high anopheline diversity, high vector abundance, changes in relative species abundance from historic data, and sustained transmission of malaria in endemic areas of Bangladesh.

Life Cycle and Ethological Notes onBurca braco braco(Herrich-Schäffer, 1865) (Hesperiidae: Pyrginae)

ABSTRACT. In this study, we describe for the first time the immature stages of a Burca species: eggs, five larval instars, prepupa, and pupa. Morphometric measurements for each stage are given. Based on the measures of head width and growth rates, we recognize the possible presence of non-typical individuals with more than five instars. We describe four larval shelters types that are associated with specific instars. Each individual constructs three to four shelters during its larval development. We also report notes on feeding patterns and oviposition.

Differentiation of movement behaviour in an adaptively diverging salamander population.

Dispersal is considered to be a species-specific trait, but intraspecific variation can be high. However, when and how this complex trait starts to differentiate during the divergence of species/lineages is unknown. Here, we studied the differentiation of movement behaviour in a large salamander population (Salamandra salamandra), in which individual adaptations to different habitat conditions drive the genetic divergence of this population into two subpopulations. In this system, salamanders have adapted to the deposition and development of their larvae in ephemeral ponds vs. small first-order streams. In general, the pond habitat is characterized as a spatially and temporally highly unpredictable habitat, while streams provide more stable and predictable conditions for the development of larvae. We analysed the fine-scale genetic distribution of larvae, and explored whether the adaptation to different larval habitat conditions has in turn also affected dispersal strategies and home range size of adult salamanders. Based on the genetic assignment of adult individuals to their respective larval habitat type, we show that pond-adapted salamanders occupied larger home ranges, displayed long-distance dispersal and had a higher variability of movement types than the stream-adapted individuals. We argue that the differentiation of phenotypically plastic traits such as dispersal and movement characteristics can be a crucial component in the course of adaptation to new habitat conditions, thereby promoting the genetic divergence of populations.

Occurrence of Anisakis (Nematoda: Anisakidae) larvae in unusual hosts in Southern hemisphere

Nematodes belonging to the genus Anisakis are important parasites due to their abundance in seafood and health impacts on humans. In the present study Anisakis larvae were found in a number of uncommon hosts including the Grey petrel, Procellaria cinerea, the Little penguin, Eudyptula minor, Blue-lipped sea krait, Laticauda laticaudata and Spinner shark, Carcharhinus brevipinna.Morphological examination showed nematodes in these animals are Anisakis larval type I. Genetic characterisation suggested that the larva from one Grey petrel was Anisakis berlandi, whereas the other larvae from the second Grey petrel and from the little penguin were Anisakis pegreffii. A number of larvae found in Blue-lipped sea krait and Spinner shark were identified as Anisakis typica. This is the first report of infective stage of Anisakis larvae parasitising hosts other than teleost fish. Understanding of the extent of infection and the pathogenicity of anisakid nematodes in hosts found in the present study is important in the conservation studies and management plans of these hosts.

NMR Profiling of Metabolites in Larval and Juvenile Blue Mussels (Mytilus edulis) under Ambient and Low Salinity Conditions

Blue mussels (Mytilus edulis) are ecologically and economically important marine invertebrates whose populations are at risk from climate change-associated variation in their environment, such as decreased coastal salinity. Blue mussels are osmoconfomers and use components of the metabolome (free amino acids) to help maintain osmotic balance and cellular function during low salinity exposure. However, little is known about the capacity of blue mussels during the planktonic larval stages to regulate metabolites during osmotic stress. Metabolite studies in species such as blue mussels can help improve our understanding of the species’ physiology, as well as their capacity to respond to environmental stress. We used 1D 1H nuclear magnetic resonance (NMR) and 2D total correlation spectroscopy (TOCSY) experiments to describe baseline metabolite pools in larval (veliger and pediveliger stages) and juvenile blue mussels (gill, mantle, and adductor tissues) under ambient conditions and to quantify changes in the abundance of common osmolytes in these stages during low salinity exposure. We found evidence for stage- and tissue-specific differences in the baseline metabolic profiles of blue mussels, which reflect variation in the function and morphology of each larval stage or tissue type of juveniles. These differences impacted the utilization of osmolytes during low salinity exposure, likely stemming from innate physiological variation. This study highlights the importance of foundational metabolomic studies that include multiple tissue types and developmental stages to adequately evaluate organismal responses to stress and better place these findings in a broader physiological context.

Oligomeric larvae of the pycnogonids revisited.

Organization and ultrastructure of the protonymphon larva were never adequately described, despite it being the common larval type of the enigmatic sea spiders and the only example of oligosegmented life stage among recent chelicerates. We have made a comprehensive examination of the newly hatched free-living protonymphons of Nymphon brevirostre using SEM, TEM, light, and confocal microscopy. Although fairly typical in their broad characters, protonymphon larvae have a number of unique and unexpected traits. Body cavity, already present at this stage, is lined with extracellular matrix and thus is conclusively identified as primary body cavity. Central nervous system includes four postocular neuromeres arranged in three ganglia: supraesophageal, subesophageal, and the first ganglion of the ventral nerve cord. Examination of the sensory organs revealed unusually organized eyes, mechanoreceptors, and chemoreceptors. We have uncovered a mixed sensory-secretory nature of chelar glands and proposed possible modalities of its receptory part. We gave first descriptions of the complex ultrastructure of three secretory organs (spinning glands, slit-like organs, proboscis glands) and hypothesized on their mode of functioning. Comparisons with another oligomeric larva, for example, nauplius, revealed discrepancies in the segmentation of these animals. Although both larvae are externally unsegmented and bear three pairs of homologous appendages, the protonymphon body includes a fourth segment of the prospective walking legs which is absent in nauplius.

Description and genetic characterization of a new Contracaecum larval type (Nematoda: Anisakidae) from Australia.

Nematode parasites belonging to the genus Contracaecum are economically important parasites with zoonotic significance. Adult Contracaecum spp. are found in the stomach of marine mammals or piscivorous birds, and larval stages infect a wide range of invertebrates and fish species. Human infection with Contracaecum larvae has been reported in Australia and other countries after the consumption of infected fish. Although the genus Contracaecum comprises numerous species, thus far only four Contracaecum larval types have been specifically identified, therefore their life cycle and biology are not yet fully understood. In this study, a new Contracaecum larva (type IV), found in the intestinal tissue of carp caught from Coonancoocabil Lagoon, New South Wales, Australia, is described and characterized genetically. It was identified as Contracaecum bancrofti, a unique Australian species reported previously from the Australian pelican in Northern Territory, New South Wales, Victoria and South Australia. This study highlights the role of migratory birds and introduced fish species, such as carp, in distributing zoonotic pathogens not only across the continent but also from marine to freshwater systems. Coonancoocabil Lagoon is located in the Murrumbidgee Valley National Park, a highly managed conservation area with native fish in wetlands located in close proximity to several fish farms and hatcheries. Infection of a highly resilient invasive fish species, such as carp, with a zoonotic parasite of low host specificity, such as Contracaecum larvae, should be alarming for aquaculture and environmental authorities.

Do larval types affect genetic connectivity at sea? Testing hypothesis in two sibling marine gastropods with contrasting larval development

In marine environments, connectivity among populations of benthic invertebrates is provided primarily by dispersion of larvae, with the duration of pelagic larval phase (PLD) supposed to represent one of the major factor affecting connectivity. In marine gastropods, PLD is linked to specific larval development types, which may be entirely intracapsular (thus lacking a pelagic dispersal), or include a short pelagic lecithotrophic or a long planktotrophic phase.In the present study, we investigated two sibling species of the cosmopolitan neogastropod genus Columbella (commonly known as dove shells): Columbella adansoni Menke, 1853, from the Macaronesian Atlantic archipelagos, with planktotrophic development, and Columbella rustica Linnaeus, 1758, from the Mediterranean Sea, with intracapsular development.We expected to find differences between these two sister species, in terms of phylogeographic structure, levels of genetic diversification and spatial distribution of genetic diversity, if PLD was actually a relevant factor affecting connectivity.By analysing the sequence variation at the cytochrome c oxidase subunit I (COI) in 167 specimens of the two species, collected over a comparable geographic range, we found that Columbella adansoni, the species with planktotrophic development, and thus longer PLD, showed no phylogeographic structure, lower levels of genetic diversity, interpopulational variance lower than the intrapopulational one and no spatial structure in the distribution of the genetic diversity; Columbella rustica, the species with intracapsular development, thus with evidently lower dispersal abilities, showed a clear phylogeographic structure, higher levels of genetic diversity, high interpopulational and low intrapopulational variance, and a clear signature of global spatial structure in the distribution of the genetic diversity.Thus, in this study, two sibling species differing almost only in their larval ecology (and PLD), when compared for their genetic variation showed patterns supporting the hypothesis that PLD is a major factor affecting genetic connectivity.Therefore, it seems reasonable to expect that the ecological attributes of the marine communities - also in terms of the variation in larval ecology of the species involved – are taken into the due consideration in conservation actions, like the design of marine protected areas networks.

Morphological and molecular characterization of selected species of Hysterothylacium (Nematoda: Raphidascarididae) from marine fish in Iraqi waters.

Hysterothylacium species are perhaps the most abundant and diverse group of marine ascaridoids; however, their life cycle and specific identification in larval stages in many parts of the world, particularly in Iraqi marine waters, have not been completely understood. In this study three members of the genus Hysterothylacium collected from Khor Abdulla in Iraq are morphologically described, genetically characterized and their relationship with other closely related taxa are compared and discussed. A new Hysterothylacium larval type in the fourth stage of development is described, and morphological and molecular evidence (based on the sequences of internal transcribed spacers) are provided for its distinction from previously known fourth-stage Hysterothylacium larval types. Based on the sequence data it is suggested that the new larval type, which herein was assigned as Hysterothylacium larval type XVI, is H. persicum which was previously reported from the close proximity in Bandar Abbas, Iran. In addition, two other taxa, including Hysterothylacium larval type XV and H. reliquens, have been found in the present study, for which new hosts are reported. This study provides some insights into the taxonomy and systematics of these parasites, not only in this region but also for similar studies elsewhere.

Inscuteable maintains type I neuroblast lineage identity via Numb/Notch signaling in the Drosophila larval brain

In the Drosophila larval brain, type I and type II neuroblasts (NBs) undergo a series of asymmetric divisions which give rise to distinct progeny lineages. The intermediate neural progenitors (INPs) exist only in type II NB lineages. In this study, we reveal a novel function of Inscuteable (Insc) that acts to maintain type I NB lineage identity. In insc type I NB clones of mosaic analyses with a repressible cell marker (MARCM), the formation of extra Deadpan (Dpn)+ NB-like and GMC-like cells is observed. The lack of Insc leads to the defective localization and segregation of Numb during asymmetric cell division. By the end of cytokinesis, this results in insufficient Numb in ganglion mother cells (GMCs). The formation of extra Deadpan (Dpn)+ cells in insc clones is prevented by the attenuation of Notch activity. This suggests that Insc functions through the Numb/Notch signaling pathway. We also show that in the absence of Insc in type I NB lineages, the cellular identity of GMCs is altered where they adopt an INP-like cell fate as indicated by the initiation of Dpn expression accompanied by a transient presence of Earmuff (Erm). These INP-like cells have the capacity to divide multiple times. We conclude that Insc is necessary for the maintenance of type I NB lineage identity. Genetic manipulations to eliminate most type I NBs with overproliferating type II NBs in the larval brain lead to altered circadian rhythms and defective phototaxis in adult flies. This indicates that the homeogenesis of NB lineages is important for the adult's brain function.

Morphogenesis and fine structure of the developing jaws of Mooreonuphis stigmatis (Onuphidae, Annelida)

The morphology of the jaw apparatus of Mooreonuphis stigmatis was studied at different stages of development using light and electron microscopy as well as 3D reconstruction techniques. Three generations of maxillary apparatuses were identified. The first (larval) type of maxillae was found in juveniles that have from 4 up to 15 chaetigers. They are formed by short microvilli and have no capacity for continuous growth, as they are completely replaced by the subsequent generation of jaws. The second (juvenile) type of maxillae appears in individuals that have from 10 to 18 chaetigers. They are fully penetrated by long microvilli and have a layer of collagen fibrils in the basicuticle. The third (adult) type of maxillae is found in worms that have at least 92 chaetigers. These maxillae have long microvilli that penetrate the intermediate layer of the basicuticle but do not reach the thick collagen layer. The mandibles and two generations of maxillae (adult and juvenile) have the capacity for continuous growth. Previously, it was thought that different types of jaw structures are characteristic of different annelid taxa (for example, Dorvilleidae, Adercodon, etc.). Our data suggest that all three types of structures are found during developmental changes of the jaws in one species. The similarity between the larval maxillae of M. stigmatis and maxillary apparatus of Dorvilleidae supports the progenetic origin of the latter. The features of fine morphology suggest that the larval maxillae of Onuphidae have more chances to be fossilized than the adult maxillae. Consequently, the larval maxillae of labidognath Eunicida could be better represented in fossil material, while the calcified adult maxillae have a lower preservation capacity.

Development of a lecithotrophic pilidium larva illustrates convergent evolution of trochophore-like morphology

BackgroundThe pilidium larva is an idiosyncrasy defining one clade of marine invertebrates, the Pilidiophora (Nemertea, Spiralia). Uniquely, in pilidial development, the juvenile worm forms from a series of isolated rudiments called imaginal discs, then erupts through and devours the larval body during catastrophic metamorphosis. A typical pilidium is planktotrophic and looks like a hat with earflaps, but pilidial diversity is much broader and includes several types of non-feeding pilidia. One of the most intriguing recently discovered types is the lecithotrophic pilidium nielseni of an undescribed species, Micrura sp. “dark” (Lineidae, Heteronemertea, Pilidiophora). The egg-shaped pilidium nielseni bears two transverse circumferential ciliary bands evoking the prototroch and telotroch of the trochophore larva found in some other spiralian phyla (e.g. annelids), but undergoes catastrophic metamorphosis similar to that of other pilidia. While it is clear that the resemblance to the trochophore is convergent, it is not clear how pilidium nielseni acquired this striking morphological similarity.ResultsHere, using light and confocal microscopy, we describe the development of pilidium nielseni from fertilization to metamorphosis, and demonstrate that fundamental aspects of pilidial development are conserved. The juvenile forms via three pairs of imaginal discs and two unpaired rudiments inside a distinct larval epidermis, which is devoured by the juvenile during rapid metamorphosis. Pilidium nielseni even develops transient, reduced lobes and lappets in early stages, re-creating the hat-like appearance of a typical pilidium. Notably, its two transverse ciliary bands can be ontogenetically linked to the primary ciliary band spanning the larval lobes and lappets of the typical planktotrophic pilidium.ConclusionsOur data shows that the development of pilidium nielseni differs remarkably from that of the trochophore, even though their larval morphology is superficially similar. Pilidium nielseni’s morphological and developmental features are best explained by transition from planktotrophy to lecithotrophy in the context of pilidial development, rather than by retention of or reversal to what is often assumed to be the spiralian ancestral larval type — the trochophore. Development of pilidium nielseni is a compelling example of convergent evolution of a trochophore-like body plan within Spiralia.

Use of agar-bound microparticulate diet as alternative food for tropical abalone, Haliotis asinina (Linnaeus 1758) post-larvae in large-scale cultures

The efficacy of using agar-bound microparticulate diet (A-MPD) as alternative food for abalone Haliotis asinina Linne post-larvae in large-scale culture was investigated. Larvae sourced from the hatchery-bred (HB) and wild-sourced (WS) broodstock were fed with either diatoms (TMT1-NF), agar-bound microparticulate diet (TMT2-A-MPD), or a combination of both feeds (TMT3-NF + A-MPD) in six 2-m3 tanks replicated over time. Three hundred thousand veliger larvae were stocked/tank containing 80 corrugated plates with mucus trails hanging on bamboo poles. Feeds were given at 0900 h starting at day 3 with seawater flow through introduced every 1400 h starting day 5. Two-way analysis of variance determined significant differences (p < 0.05) in survival and shell length between larval sources and feed types. Tukey’s post hoc test established differences among treatment means. At day 30, survival for HB- and WS-sourced larvae was significantly higher (42%) in TMT3 compared with TMT2 having 35% for HB and 38% for WS (p < 0.05). Larvae fed with TMT1 had significantly lowest survival among the three treatments. Survival at 60 and 90 days did not show significant difference for TMT2 and TMT3 regardless of broodstock source. Post-larval shell growth (90 days), from both sources fed with TMT2 and TMT3, was significantly higher than TMT1 (p < 0.05). Larval performance did not show any significant interactions between HB and WS broodstock. The use of A-MPD alone or in combination may elicit improvement in survival and shell length growth in abalone larvae regardless of larval sources. A-MPD may be used as full or partial replacements to diatoms as alternative food for abalone post-larvae in large-scale culture.

Epigenetic modifications and their relation to caste and sex determination and adult division of labor in the stingless bee Melipona scutellaris.

Stingless bees of the genus Melipona, have long been considered an enigmatic case among social insects for their mode of caste determination, where in addition to larval food type and quantity, the genotype also has a saying, as proposed over 50 years ago by Warwick E. Kerr. Several attempts have since tried to test his Mendelian two-loci/two-alleles segregation hypothesis, but only recently a single gene crucial for sex determination in bees was evidenced to be sex-specifically spliced and also caste-specifically expressed in a Melipona species. Since alternative splicing is frequently associated with epigenetic marks, and the epigenetic status plays a major role in setting the caste phenotype in the honey bee, we investigated here epigenetic chromatin modification in the stingless bee Melipona scutellaris. We used an ELISA-based methodology to quantify global methylation status and western blot assays to reveal histone modifications. The results evidenced DNA methylation/demethylation events in larvae and pupae, and significant differences in histone methylation and phosphorylation between newly emerged adult queens and workers. The epigenetic dynamics seen in this stingless bee species represent a new facet in the caste determination process in Melipona bees and suggest a possible mechanism that is likely to link a genotype component to the larval diet and adult social behavior of these bees.