Early Juvenile Stages Research Articles

Light and scanning electron microscope observations of the digenean intestinal parasite Acanthostomum spiniceps Looss 1896 (Cryptogonimidae) from the catfish Bagrus bajad and B. docmak

Fishes of the River Nile are a promising source of animal protein. The catfishes Bagrus bajad Fabricius 1775 and Bagrus docmak Fabricius 1775 are economically important with well-marketable size and use in fish farming. Digenean parasites cause severe damage to the gut tissue of their fish hosts. There are still some questions regarding the taxonomical features of the genus Acanthostomum Looss 1899. The present study has revealed new morphological features that confirmed the identity of the collected worms as Acanthostomum spiniceps Looss 1896 and aided in elucidating the possible functions of different internal organs and surface features. A long-stemmed excretory bladder was detected but there was no evidence of gonotyl. The circumoral spines (28–30) were detected in the early juvenile stage and adult. The ventral sucker has no tegumental folds in juveniles or crescent-shaped tegumental folds in adults but possesses two overlapping lips. In both juveniles and adults, the oral sucker with the associated circumoral spines assumed retracted, and protracted positions reflecting its highest movement activity. Elongation of the anterior region and partial emergence of the oral cavity in living juveniles and adults were described. The possible functions of the forebody glands and the posterior body openings near the anal pores were discussed. Large, non-ciliated, dome-shaped, and small ciliated, button-like papillae were concentrated on the body surface, particularly the oral and ventral suckers. The tegument possesses densely arranged single-pointed and scale-like spines that gradually decrease in size and number as they proceed posteriorly.

Bacterivory of the hydrothermal-vent-specific copepod Stygiopontius senokuchiae (Dirivultidae, Siphonostomatoida) from copepodite through adult stages

Abstract Stygiopontius senokuchiae is a hydrothermal-vent-specific copepod species (Dirivultidae, Siphonostomatoida) and abundantly distributes near vent orifices. This species is thought to have an early juvenile planktic stage, and previous studies using bulk tissue stable-isotope and radioisotope data suggest that adults ingest chemoautotrophic microbes growing on vent chimneys. However, ontogenetic changes in their diets have not been investigated. We analysed gut contents of copepodite I to adult stages of S. senokuchiae collected from a hydrothermal-vent chimney at the Izu-Ogasawara Arc to check for the presence of bacterial cells in oral tubes and guts. We compared these results with an unidentified co-occurring calanoid and a species of Ectinosoma (Harpacticoida) and to other Siphonostomatoid copepods, namely Hatschekia labracis (Hatschekiidae), which was attached to a scarbreast tuskfin (Choerodon azurio, Labridae), and Asterocheres sp. 1 (Asterocheridae), which was attached to a spirastrellid sponge. Carbon isotope ratios of S. senokuchiae at different stages and Ectinosoma were measured to complement the nutritional insights obtained from gut-content analysis. Our results clearly showed that most S. senokuchiae individuals possessed bacteria in the oral tube or gut regardless of growth stage, whereas the coexisting Ectinosoma and calanoid did not. The carbon isotopic compositions confirmed that S. senokuchiae gains its nutrition from chemoautotrophic bacteria that use the rTCA carbon-fixation pathway. Comparisons with other Siphonostomatoida copepods suggest that Dirivultidae are specifically adapted to feed on bacteria at hydrothermal-vent chimneys, allowing their high dominance and evolutionary success in these habitats.

Exogenous myo-inositol increases salt tolerance and accelerates CAM induction in the early juvenile stage of the facultative halophyte Mesembryanthemum crystallinum but not in the late juvenile stage.

Mesembryanthemum crystallinum L. (ice plant) develops salt tolerance during the transition from the juvenile to the adult stage through progressive morphological, physiological, biochemical, and molecular changes. Myo-inositol is the precursor for the synthesis of compatible solute D-pinitol and promotes Na+ transport in ice plants. We previously showed that supplying myo-inositol to 9-day-old seedlings alleviates salt damage by coordinating the expression of genes involved in inositol synthesis and transport, affecting osmotic adjustment and the Na/K balance. In this study, we examined the effects of myo-inositol on physiological parameters and inositol-related gene expression in early- and late-stage juvenile plants. The addition of myo-inositol to salt-treated, hydroponically grown late juvenile plants had no significant effects on growth or photosynthesis. In contrast, supplying exogenous myo-inositol to salt-treated early juvenile plants increased leaf biomass, relative water content, and chlorophyll content and improved PSII activity and CO2 assimilation. The treatment combining high salt and myo-inositol synergistically induced the expression of myo-inositol phosphate synthase (INPS), myo-inositol O-methyltransferase (IMT), and inositol transporters (INTs), which modulated root-to-shoot Na/K ratio and increased leaf D-pinitol content. The results indicate that sufficient myo-inositol is a prerequisite for high salt tolerance in ice plant.

Knockout of microphthalmia-associated transcription factor (mitf) confers a red and yellow tilapia with few pigmented melanophores

As the most important genetic factor for melanophore differentiation and melanogenesis, mitf has been studied for over 30 years. The phenotypes of mitf mutants have also received continuous attention for nearly a century. Due to the third round of genome duplication, teleosts have two mitf genes. Mutation analysis demonstrates that mitfa plays an important role in zebrafish pigmentation. However, there have been no functional studies on body color regulation by mitfb or studies of mitfa;mitfb double mutants. In the present study, we mutated both mitf genes in tilapia using CRISPR/Cas9. Disruption of mitfa resulted in light yellow body color with weak gray vertical bars due to significantly reduced numbers of melanophores and increased sizes of xanthophores, while disruption of mitfb led to slight hypo-pigmentation. Double mutation of mitfa and mitfb resulted in dramatic hypo-pigmentation in trunk and fins due to loss of additional melanophores and increase in the number of iridophores compared to the single mutants, but had no influence on retinal pigment epithelium (RPE) pigmentation. The mitfa−/−;mitfb−/− mutants were yellow with black spots at early juvenile stage (60 dpf), yellow-reddish at late juvenile stage (120 dpf) and red and yellow (including iris) at adult stage (180 dpf), due to increased erythrophores and enlarged xanthophores. Our results demonstrated that both mitf genes are important for body color formation in tilapia, but mitfa plays a more important role than mitfb. Additionally, by comparing the phenotypes of the mitfa−/−;mitfb−/−, pmela−/−;pmelb−/− and hps4−/− mutants, we found that disruption of genes with different functions in different aspects of melanogenesis could lead to different body colors in tilapia. Mitf is the most important gene for melanophore differentiation and is probably also critical for differentiation of erythrophores, xanthophores and iridophores. To our knowledge, this is the first report showing that both mitf genes are involved in body color formation by loss of function study, and the first report showing that erythrophore numbers and xanthophore sizes were affected by mitf. Our research not only serves as a model for studying mitf function in tilapia and the closely related cichlids, but also provides new strategies for breeding red and yellow tilapia for aquaculture.

Differences in the Morphology and Reproduction of the Ascidian Boltenia Villosa Across a Latitudinal Gradient

While the larval and early juvenile stages of the stalked ascidian Boltenia villosa are well-documented in the literature, little is known about the range of morphological variation in B. villosa adults. Anecdotal evidence suggests that larger individuals with shorter stalks make up the populations found in Washington, while Oregon populations consist of smaller individuals with longer stalks. The present study aimed to develop a qualitative understanding of the morphological and reproductive differences across the latitudinal gradient of B. villosa. Morphological differences between populations were studied through a combination of morphometrics and scanning electron microscopy. To compare reproductive outputs, the diameters of oocytes and the gonadosomatic indices of representative individuals were measured. The results suggest significant differences between the two populations in body proportions, spine character, and reproduction despite their genetic similarities. These preliminary results provide the basis for future research into the distribution of B.villosa and the possible existence of an undescribed subspecies of B. villosa.

Influence of larval prey availability on year class strength in Faroe Plateau cod (Gadus morhua)

Recruitment to the Faroe Plateau cod (Gadus morhua) stock (62°N, 6.8°W) is highly variable between years, and availability of suitable prey during the early life stages may determine the year class strength. Here we first investigate interannual variability in recruitment (1983–2017) of cod based on survey data and data from stock assessments. Positive relationships between the spawning stock biomass and larval abundance and between juvenile abundance and recruitment at year 1 were detected. However, the relationship between larval abundance and juvenile abundance was not statistically significant, indicating variable survival during the larval/early juvenile stages. We then investigated if food availability at the larval and early juvenile stages could explain the variability in recruitment using an 8-year long time series of larval and juvenile gut content. Feeding at the early larval stage, present during the pre-bloom, was relatively stable between years. However, large interannual variability in feeding at the early juvenile stage was detected. When juvenile survival and mean length were high, juveniles clearly had less food in their guts. This contradiction is suggested to be due to intense grazing pressure on the zooplankton at high juvenile abundances. However, years with high larval survival coincided with years when the phytoplankton spring bloom was early and intense, suggesting that production of suitable prey during the larval stages determines larval growth and survival and that this production is dependent on the phytoplankton spring bloom characteristics.

Deletion of eif6 Partially Rescues Survival, but Not Neutropenia, of Zebrafish Model of Shwachman-Diamond Syndrome: The Roles of Sbds and Eif6 in Perturbing Lipid Metabolism

Introduction: Shwachman-Diamond syndrome (SDS) is an autosomal recessive disorder characterized by pancreatic insufficiency, skeletal defects, neutropenia, and an increased risk of myelodysplastic syndrome (MDS)/acute myeloid leukemia (AML). Biallelic mutations in SBDS account for ~90% of patients. SBDS interacts physically with EFL1 to release EIF6 from the pre-60S ribosomal subunit and promote the assembly of the mature 80S ribosome in the cytoplasm. A subset of SDS patients acquire del(20q), resulting in loss of EIF6. Additionally, somatic heterozygous EIF6 mutations have been found in SDS patients. SDS patients harboring del(20q) and EIF6 mutations are linked to a lower risk of developing MDS/AML when compared to other SDS patients. Together these factors point to a potential mechanism whereby a reduction in EIF6 levels may lead to a suppression in ribosome biogenesis defects. Methods and Results: We have created sbds knockout (KO) zebrafish strains that phenocopy the human syndrome with stunted growth, neutropenia and atrophy in the pancreas, liver, and intestine. Unlike the Sbds-null mice which die in early embryogenesis, sbds-null fish survive to early juvenile stages. The sbds KO fish display an accumulation of Eif6 in liver and intestine. Interestingly, we also found an accumulation of EIF6 in SDS patient-derived lymphoblasts. EIF6 has been described as a key regulator of metabolism, specifically stimulating glycolytic and fatty acid synthesis. Lipid metabolomics of sbds KO fish show a decrease in free fatty acids and phosphatidylcholine. Expression of several genes critical in lipid metabolism (srebp1, fasn and pparg) is upregulated in sbds KO fish. To study the role of Eif6 in the pathophysiology of SDS, we created an eif6 KO zebrafish with an insertion of one base pair resulting in a premature stop codon. By immunoblotting we demonstrated that mutants did not display Eif6 by 6 days post fertilization (dpf), and heterozygous fish displayed less protein than wildtype siblings. This new strain showed early mortality (~10 dpf), neutropenia, tp53 pathway activation, with a pronounced upregulation of cdkn1a as early as 5 dpf. RNA-Seq data of eif6 KO fish at 5 dpf showed a total of 2225 differentially expressed genes (DEGs): 668 downregulated and 1557 upregulated. These DEGs are involved in two main gene expression pathways: ribosome biogenesis and oxidative phosphorylation. We also observed a dysregulation of lipid metabolism markers in the eif6 KO fish. Furthermore, we bred the sbds KO with the eif6 KO, we observed a partial rescue on survival only in sbds-/- with one copy of eif6(eif6+/-) but failed to rescue their neutropenia. Conclusions: Our organismal models of sbds or eif6 deletion provide new insights into the pathophysiology of human SDS: 1) SBDS affects lipid metabolism possibly due to an accumulation of EIF6, 2) Loss of eif6 affects development/survival at an earlier stage than loss of sbds, and 3) Loss of either sbds or eif6 markedly upregulates cdkn1a, which is downstream of tp53. Interestingly, Eif6 partially rescues survival of sbds-null organisms, but only in the haploinsufficient state. EIF6 may offer a promising target for a novel therapeutic strategy in SDS.

Predators and habitat association of post-settlement snapper (Chrysophrys auratus).

Structured habitats play an important nursery role during the crucial early juvenile or post-settlement stages of many fish species. Predominantly, the utility of structured habitats to juvenile fish is thought to be associated with the provisioning of food or as a refuge from predation. Although snapper (Chrysophrys auratus) in New Zealand also have a strong affinity for structured habitats during their post-settlement phase, their predators are unknown as is the role of predation in determining habitat association. Here the authors investigated potential predators of post-settlement snapper by remotely observing interactions of restrained post-settlement snapper with potential predators and investigating the diet of potential predators. They also conducted tank experiments with a potential predator, both with and without the presence of structure. Restrained snapper were infrequently approached by predators, but two new nocturnal predators were identified. No snapper were observed in the diet of potential predators, although two piscivores were identified as potential candidates. No predation occurred during tank experiments, but there was a non-significant indication that under threat of predation post-settlement snapper may use habitat when it is present and aggregate together when it is not. The findings suggest that the pulsed nature of predation may have made it difficult to observe given the methods employed and that the threat of predation may be sufficient to drive the habitat selection of post-settlement snapper. Investigating the significance of predation via methods that do not require direct observations may therefore be more appropriate given this context.

Persistent high hatchery recruitment despite advanced reoligotrophication and significant natural spawning in a whitefish

Many lakes of the pre-Alpine region suffered from severe eutrophication that affected the natural reproduction of whitefish (Coregonus spp.) and necessitated large-scale supportive breeding programs. With the advanced reoligotrophication, it is now important to evaluate the relevance of continued artificial breeding for population dynamics. We focused on a whitefish population of a lake that has reached phase III of the reoligotrophication, i.e., lake biomass production is declining since 2012 in response to low phosphorus concentrations. We show that most eggs are naturally spawned, the observed oxygen concentrations would again support embryo development at all depths, and ready-to-hatch embryos can indeed be found on spawning grounds. We marked all hatchery-produced eggs of the 2014 cohort with Alizarin red, stocked them as usual (at larval or early juvenile stages), and recaptured them over a period of five years. Fish were aged from yearly growth rings on scales, and otoliths were checked for marks. We found 90.3 % of the 2014 cohort to be hatchery-born. This ratio did not decline with fish age. We also determined juvenile growth of the cohorts 2012–2020 (based on the first annual ring on scales) and found that stocking intensity predicted juvenile growth (r2 = 0.67). This strong density dependence of individual growth suggests that stocking has largely determined cohort size over the first nine years of reoligotrophication phase III. We conclude that large areas of spawning grounds allow again for successful embryogenesis, that large quantities of eggs are naturally spawned, but that natural recruitment is significantly reduced by ecological or evolutionary factors, e.g., competition with hatchery-born fish, desynchronization of trophic interactions, or long-term effects of fishing- or hatchery-induced evolution.

Distribution, Biology and Ontogeny of Indian River Shad (Tenualosa ilisha)

The distribution, biology and ontogenic development of the Indian River Shad (Tenualosa ilisha) compared with some other fishes compile in this review. The distribution in the inland waters of Bangladesh, stock/race, movement and migration pattern, size and sex ratio, age of reproduction, maturity and fecundity and ontogenic development of the mouth, digestive tract and accessory gland of T. ilisha discussed in this article. The life cycle of a fish starts after fertilization, but the mouth and digestive tract development start after hatching. Yolk sac larvae (3.83 ± 0.57 mm SL), pre flexion larvae (8.07 ± 1.83 mm SL), flexion larvae (11.50 ± 1.55 mm SL), post flexion larvae (14.72 ± 1.70 mm SL), and juvenile (18.75 ± 3.02 mm SL) were separated into five groups based on morphological characteristics. At the yolk sac stage, the mouth was closed, however at the pre-flexion stage, the larval mouth gape varied between 177 ± 25 and 367 ± 47 µm. The ranges were 241 ± 31 and 497 ± 59 µm, for the flexion stage, 307 ± 38 and 621 ± 74 µm for the post-flexion stage, and 393 ± 61 and 788 ± 119 µm for the early juvenile stage. Development of the digestive tract is complete at the juvenile stage of this species.

Taphonomy of drought afflicted tetrapods in the Early Triassic Karoo Basin, South Africa

The sedimentology and taphonomy of in-situ fossils from earliest Triassic strata (Induan) in the southern Karoo Basin of South Africa is presented as evidence for episodes of drought-induced mass death of the resident tetrapods. Abundant skeletons are preserved in a 2 m-thick tabular silty-sandstone capping a multi-storeyed low-sinuosity channel sandstone interpreted as a wide shallow channel that became progressively abandoned, with more ephemeral flow regime than in the underlying channels and subjected to intermittent flows of low-density sediment-laden floodwaters. Stratigraphic and planimetric distribution of 170 in-situ tetrapod fossils shows several clusters of up to eight closely-spaced articulated Lystrosaurus skeletons preserved in prone and spread-eagled body position. These are interpreted as drought-stricken carcasses that collapsed and died of starvation in and alongside dried-up water sources. Two of the specimens display an unusual micritic envelope with a distinctive pustular texture interpreted as permineralised mummified skin indicative of rapid desiccation after death. Bonebeds of disarticulated bones of multiple juvenile Lystrosaurus occur in shallow depressions within the rubified mudstones. Layering of different skeletal elements suggests some hydraulic sorting but the initial aggregation was likely a behavioural response to drought. Osteohistology of spread-eagled Lystrosaurus (L. declivis and L. murrayi species) skeletons show that they represent early juvenile stage which is in accordance with previous findings that throughout Pangaea Early Triassic Lystrosaurus died relatively young due to environmental stressors. Our results support the hyperthermal hypothesis that ∼252 Mya increased continental aridity, already a consequence of the coalescence of Pangaea, was critically intensified by volcanogenic greenhouse gasses from the Siberian traps. We propose that in the aftermath of the End-Permian mass extinction event, a succession of climatic drying episodes orchestrated a series of fully-functioning terrestrial ecosystems that were markedly different to those of the pre-extinction, and likely had a profound and lasting influence on the evolution of tetrapods.

DHA-enrichment of live and compound feeds influences the incidence of cannibalism, digestive function, and growth in the neotropical catfish Pseudoplatystoma punctifer (Castelnau, 1855) during early life stages

Pseudoplatystoma punctifer is a highly appreciated fish species native to the Amazon basin, whose commercial farming has been hampered by low survival during early life stages due to the high incidence of cannibalism and the low acceptability of compound diets at weaning. Dietary DHA is known to promote digestive system development and maturation and growth, whereas its deficiency induces stress. The aim of this study was to evaluate the effect of dietary DHA supplementation on the incidence of cannibalism, digestive physiology, and growth performance during the early life stages of P. punctifer to improve current feeding protocols. Four dietary treatments were generated using a commercial enrichment product with high DHA content. Fish larvae were fed non-enriched or enriched Artemia from 4 to 15 days post fertilization (dpf) and fed a non-enriched or enriched compound diet from 15 to 26 dpf, coinciding with the start of the juvenile stage. Growth, survival, incidence of cannibalism, proximate and fatty acid composition, histology of the intestine and liver, and quantitative gene expression of the main digestive enzymes (amy, try, ctr, pga, pla2, and lpl) were analyzed in the different dietary groups at the end of each feeding period. Results showed that dietary DHA supplementation influenced P. punctifer in a developmental stage-dependent manner. In particular, DHA-enriched Artemia provided during the larval stage contributed to reduce the incidence of cannibalism and improved survival at the early juvenile stage, while enriching the compound diet improved growth. The expression level of genes involved in protein and carbohydrate digestion (ctr, amy) was higher in groups fed enriched Artemia despite that both enriched and non-enriched Artemia displayed similar proximate composition, suggesting that different dietary fatty acid profiles may modulate the expression of these digestive enzyme precursors. The group transitioning from enriched Artemia to non-enriched compound diet showed a higher expression of most genes at the early juvenile stage. At the histological level, the group fed non-enriched Artemia and compound diet showed a significant accumulation of lipids in the intestine relative to the liver, contrary to the rest of the groups that showed similar amounts of lipids in both tissues, indicating a more balanced lipid metabolism. The group fed both DHA-enriched Artemia and compound diet provided the best results in terms of growth, survival, incidence of cannibalism, and digestive physiology. In conclusion, this study showed that the nutritional history during the larval period affected fish nutrition and behavior during the early juvenile stage.

Impact of squid predation on juvenile fish survival

Predation is a major source of mortality during the early life stages of marine fishes; however, few studies have demonstrated its impact—especially that of squid predation—on survival processes. Here, we examined the feeding habits and predation impacts of swordtip squid on a major prey fish, juveniles of jack mackerel, in the East China Sea. Otoliths of the juveniles extracted from the squid stomach were used to reconstruct the age–length relationship and the growth trajectory of the consumed juveniles, and they were compared to those of juveniles collected with a net using a newly developed statistical framework. The juveniles consumed by squid had significantly shorter body lengths and smaller body sizes during the late larval and early juvenile stages than the netted juveniles, suggesting that smaller juveniles with slower growth rates have a higher probability to be selected. The body mass ratio of the predator squid to prey juveniles (predator–prey mass ratio, PPMR) ranged from 7 to 700, which was remarkably lower than the PPMR reported in various marine ecosystems based on analyses of fishes. Our findings demonstrate that squid predation can significantly impact the early life survival of fish and the trophodynamics in marine ecosystems.

A Baseline for Skeletal Investigations in Medaka (Oryzias latipes): The Effects of Rearing Density on the Postcranial Phenotype.

Oryzias latipes is increasingly used as a model in biomedical skeletal research. The standard approach is to generate genetic variants with particular skeletal phenotypes which resemble skeletal diseases in humans. The proper diagnosis of skeletal variation is key for this type of research. However, even laboratory rearing conditions can alter skeletal phenotypes. The subject of this study is the link between skeletal phenotypes and rearing conditions. Thus, wildtype medaka were reared from hatching to an early juvenile stage at low (LD: 5 individuals/L), medium (MD: 15 individuals/L), and high (HD: 45 individuals/L) densities. The objectives of the study are: (I) provide a comprehensive overview of the postcranial skeletal elements in medaka; (II) evaluate the effects of rearing density on specific meristic counts and on the variability in type and incidence of skeletal anomalies; (III) define the best laboratory settings to obtain a skeletal reference for a sound evaluation of future experimental conditions; (IV) contribute to elucidating the structural and cellular changes related to the onset of skeletal anomalies. The results from this study reveal that rearing densities greater than 5 medaka/L reduce the animals’ growth. This reduction is related to decreased mineralization of dermal (fin rays) and perichondral (fin supporting elements) bone. Furthermore, high density increases anomalies affecting the caudal fin endoskeleton and dermal rays, and the preural vertebral centra. A series of static observations on Alizarin red S whole mount-stained preural fusions provide insights into the etiology of centra fusion. The fusion of preural centra involves the ectopic formation of bony bridges over the intact intervertebral ligament. An apparent consequence is the degradation of the intervertebral ligaments and the remodeling and reshaping of the fused vertebral centra into a biconoid-shaped centrum. From this study it can be concluded that it is paramount to take into account the rearing conditions, natural variability, skeletal phenotypic plasticity, and the genetic background along with species-specific peculiarities when screening for skeletal phenotypes of mutant or wildtype medaka.

Nervous necrosis virus titration and antigen quantitation by indirect sandwich enzyme linked immunosorbent assay

Viral nervous necrosis (VNN) is a serious disease of marine and brackishwater fishes caused by nervous necrosis virus (NNV) resulting in up to 100% mortality in early larval and juvenile stages. Adult fish when infected are asymptomatic and spread the virus vertically to the offspring through milt and eggs. Prevention of vertical transmission of the disease is by using disease free broodstock and vaccinating the brooders. Estimation of antigen content and virus titre is essential to determine antigen/virus concentration in VNN vaccine. A monoclonal antibody based indirect sandwich ELISA was developed to quantify the NNV antigen and to estimate the virus titre by TCID50 coupled ELISA. Mouse hybridoma clones secreting monoclonal antibodies (MAb) against the capsid protein of NNV was developed and characterised. The antibodies reacted specifically with the recombinant capsid protein and purified virus in western blot. Polyclonal antibodies against NNV were used as capture antibodies and MAbs were used as detection antibodies to optimise an indirect sandwich ELISA to detect and quantify capsid protein of NNV. The developed assay had a sensitivity of 390 ng/ml and could detect the virus in clinical samples. The assay coupled with TCID50 could be used to estimate the virus titre rather than by observing the CPE which is laborious and subjective.

Developmental Temperature Shapes the Otolith Morphology of Metamorphosing and Juvenile Gilthead Seabream (Sparus aurata Linnaeus, 1758)

Otolith morphological variability is used as a reliable indicator to discriminate fish that experience different environmental conditions during their lifetimes. The present study examined the effects of developmental temperature (DT) during the egg and yolk-sac larval period on the otolith shape and asymmetry of Gilthead seabream in the later metamorphosis (56–58 days post-hatching, dph) and the early juvenile stage (93–95 dph). The experimental populations were reared at different water temperatures (17, 20, or 23 °C DT) from epiboly onset to the end of the yolk-sac larval stage (5–7 days post-fertilization, dpf) and then at a common rearing temperature (20 °C), up to the end of the trials (93–95 dph). Otolith shape and bilateral asymmetry were analyzed at metamorphosis (20–21 mm standard length, SL) and the early juvenile stage (31–32 mm SL). The results of elliptic Fourier analysis showed that DT significantly affected the otolith shape at both stages examined. Furthermore, elevated DT significantly increased the asymmetry levels of seabream otoliths in the early juvenile stage. The results are discussed in terms of the thermally induced long-term changes of seabream otolith morphology and the potential effects of the raised otolith asymmetry on wild seabream juveniles.

Genetic architecture of variation in Arabidopsis thaliana rosettes.

Rosette morphology across Arabidopsis accessions exhibits considerable variation. Here we report a high-throughput phenotyping approach based on automatic image analysis to quantify rosette shape and dissect the underlying genetic architecture. Shape measurements of the rosettes in a core set of Recombinant Inbred Lines from an advanced mapping population (Multiparent Advanced Generation Inter-Cross or MAGIC) derived from inter-crossing 19 natural accessions. Image acquisition and analysis was scaled to extract geometric descriptors from time stamped images of growing rosettes. Shape analyses revealed heritable morphological variation at early juvenile stages and QTL mapping resulted in over 116 chromosomal regions associated with trait variation within the population. Many QTL linked to variation in shape were located near genes related to hormonal signalling and signal transduction pathways while others are involved in shade avoidance and transition to flowering. Our results suggest rosette shape arises from modular integration of sub-organ morphologies and can be considered a functional trait subjected to selective pressures of subsequent morphological traits. On an applied aspect, QTLs found will be candidates for further research on plant architecture.

The neuropeptide Pth2 modulates social behavior and anxiety in zebrafish

SummaryBehavior is context-dependent and often modulated by an animal’s internal state. In particular, different social contexts can alter anxiety levels and modulate social behavior. The vertebrate-specific neuropeptide parathyroid hormone 2 (pth2) is regulated by the presence of conspecifics in zebrafish. As its cognate receptor, the parathyroid hormone 2 receptor (pth2r), is widely expressed across the brain, we tested fish lacking the functional Pth2 peptide in several anxiety-related and social behavior paradigms. Here, we show that the propensity to react to sudden stimuli with an escape response was increased in pth2−/− zebrafish, consistent with an elevated anxiety level. While overall social preference for conspecifics was maintained in pth2−/− fish until the early juvenile stage, we found that both social preference and shoaling were altered later in development. The data presented suggest that the neuropeptide Pth2 modulates several conserved behaviors and may thus enable the animal to react appropriately in different social contexts.

Recovery of Haemal Lordosis in European Seabass Dicentrarchus labrax (Linnaeus 1758)

Lordosis of the haemal part of the vertebral column is a frequent abnormality in reared fish. Haemal lordosis develops during the late larval and early juvenile life stages of fish, mainly due to the high swimming activity of the fish in the rearing tanks. In the present study, we have examined whether haemal lordosis recovers during the growth of European seabass. Furthermore, we aimed to develop simple morphometric indices (PrAn1 and PrAn2) that might link the severity of lordosis at the juvenile stage with fish morphological quality at harvesting. At 111 days post-hatching (dph, 53 ± 4 mm standard length, SL), 600 seabass juveniles with lordotic (L, 200 fish) or normal (N, 400 fish) external morphology were selected and introduced in a common tank. At 150 dph (75 ± 7 mm SL), 350 fish were randomly selected, pit-tagged and transferred to a sea cage for on-growing up to 502 dph (234 ± 16 mm SL). The morphological examination of the fish at 150 and 502 dph revealed that 60% (46 out of 77) of L juveniles turned into normal phenotype by the end of on-growing period. Interestingly, 56% of the fish with recovered external morphology (N-Rec) presented either a completely normal vertebral column (31%) or minor abnormalities of individual vertebrae (25%). Following the results of geometric morphometric analysis, the differences in the body shape between N-Rec and N fish were not statistically significant (p > 0.05, canonical variate analysis). The examined morphometric indices were effective in discriminating the normal fish from 58% (PrAn1) to 65% (PrAn2) of lordotic juveniles. Results are discussed with respect to the mechanism of lordosis recovery, and spotlights on their application for quality control and cull out of the abnormal fish in commercial hatcheries.

Notes on the morphological changes in the oocyte of the polychaete Perinereis wilsoni (Glasby and Hsieh 2006) from spawning to the early larval stage

ABSTRACT Perinereis wilsoni is a nereidid polychaete species with segments that are completely separated from each other by a septum. Recent molecular studies on this species discussed the mechanism of male and female germ cell development and germ cell regeneration. Although several molecular studies have been conducted on this species, its embryonic and larval development have not yet been described. To address this, oocyte development from spawning to the early juvenile stage was monitored. Morphological changes in relation to minutes/hours post-fertilization are described. We found several changes in the oocyte egg envelope before the start of cell division. During embryonic development, the larvae left the jelly layer as a 3-segmented nectochaete possessing a complete adult body plan, typical of the direct larval development in this species. The dorsal pygidial cirri were observed to develop fully during the five-segmented nectochaete stage. This paper provides important information on an aspect of the reproductive biology of this species.